WO2013183523A1 - Method for producing extract from lichen belonging to genus usnea - Google Patents

Method for producing extract from lichen belonging to genus usnea Download PDF

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Publication number
WO2013183523A1
WO2013183523A1 PCT/JP2013/064950 JP2013064950W WO2013183523A1 WO 2013183523 A1 WO2013183523 A1 WO 2013183523A1 JP 2013064950 W JP2013064950 W JP 2013064950W WO 2013183523 A1 WO2013183523 A1 WO 2013183523A1
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WIPO (PCT)
Prior art keywords
extract
ogase
treatment
content
lichen
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PCT/JP2013/064950
Other languages
French (fr)
Japanese (ja)
Inventor
剛史 大場
大樹 村瀬
麻実子 菊池
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花王株式会社
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Publication date
Priority claimed from JP2012130797A external-priority patent/JP2013253058A/en
Priority claimed from JP2012130799A external-priority patent/JP2013253060A/en
Priority claimed from JP2012130798A external-priority patent/JP2013253059A/en
Application filed by 花王株式会社 filed Critical 花王株式会社
Priority to CN201380019745.6A priority Critical patent/CN104244961A/en
Publication of WO2013183523A1 publication Critical patent/WO2013183523A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A61K8/368—Carboxylic acids; Salts or anhydrides thereof with carboxyl groups directly bound to carbon atoms of aromatic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/09—Lichens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9733—Lichens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH

Definitions

  • the present invention relates to a method for producing a lichen extract of the genus Usnea .
  • Endothelin is a peptide hormone derived from endothelial cells and acts on various cells and tissues through receptors. For example, it is known to cause an increase in intracellular calcium concentration in vascular smooth muscle cells and the like (Non-Patent Document 1). In recent years, endothelin has promoted an increase in intracellular calcium concentration in epidermal melanocytes (melanocytes), promoted cell proliferation via intracellular signal transduction system, and enhanced the activity of tyrosinase, the rate-limiting enzyme of melanin synthesis. has been reported (see Non-Patent Document 2).
  • endothelin is one of the melanocyte activators produced by epidermal keratinocytes (keratinocytes) (Non-patent Document 3), and is an important factor in ultraviolet-induced pigmentation and senile pigment spot formation. It has been reported (Non-Patent Documents 4 and 5).
  • a substance capable of suppressing the action of endothelin due to the biological action of endothelin suppresses melanin production and pigmentation and is useful as a whitening agent.
  • Patent Documents 1 to 3 it is known that lichen extracts of the genus Sarogase belonging to the family Parmeliaceae have an inhibitory action on melanin production (see Patent Documents 1 to 3 and Non-Patent Document 6).
  • Patent Documents 1 to 3 as a method for producing a melanin production inhibitor, lichen is cultured, and the culture is freeze-dried and then subjected to solvent extraction at a temperature of 0 ° C. to 15 ° C. .
  • the present invention relates to a method for producing an extract containing Salusose lichen belonging to a low content of usnic acid, wherein the lichenaceae lichen is extracted using an alcohol aqueous solution having an alcohol content of 80% by volume or less.
  • the present invention relates to an endothelin action inhibitor comprising as an active ingredient the extract of the genus Sarhogase lichen obtained by the above production method.
  • the present invention also relates to a whitening agent comprising, as an active ingredient, an extract from the genus Sarugagase obtained by the production method.
  • the method for producing the Ususic acid low-containing Salogase lichen extract, the endothelin action inhibitor, and the whitening agent are collectively referred to as the first embodiment of the present invention.
  • the present invention relates to a method for producing an extract of Sarugagase lichen having a high content of difractic acid and a low content of usnic acid, wherein the lichenaceae lichen is extracted using an alcohol aqueous solution having an alcohol content of 30% by volume to 80% by volume.
  • the present invention relates to a method for producing an endothelin action inhibitor, which extracts Saruogase lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume.
  • the present invention relates to a whitening agent comprising, as an active ingredient, a Sargoses genus lichen extract or endothelin action inhibitor obtained by the production method. (Hereinafter, the manufacturing method of the extract containing high amount of difractic acid and low content of usnic acid, the method of manufacturing the endothelin action inhibitor, and the whitening agent are collectively referred to as the second embodiment of the present invention.)
  • the present invention is a method of extracting Sarugagase lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment. It is the manufacturing method of the extract which performs the above process, Comprising: It is related with the manufacturing method of the Ususic acid low content Salogase genus lichen extract which the residual rate of the usnic acid after the said processing is less than 50%.
  • the present invention is a method of extracting Sarugagase lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment.
  • the present invention relates to a method for producing an extract of Lichenaceae lichen with a high content of difractic acid and a low content of usnic acid.
  • the present invention is a method of extracting Sarugagase lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment. It is related with the manufacturing method of the endothelin action inhibitor which performs the above process.
  • the present invention relates to a whitening agent comprising, as an active ingredient, a Sargoses genus lichen extract or endothelin action inhibitor obtained by the production method.
  • the present invention is meant to include all of the first embodiment, the second embodiment, and the third embodiment.
  • An object of the present invention is to provide a method for producing a Sargogase lichen extract or an endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent.
  • the inventors of the present invention have conducted intensive investigations on the components contained in the extract of Lichenaceae lichens, the search for whitening components from Lichenaceae lichen extracts, and the extraction methods that can contain these components in high concentrations.
  • difractic acid which is contained in the extract of the genus Sarhogase and represented by the following formula, has an inhibitory effect on the increase in calcium (ion) concentration in melanocytes caused by the action of endothelin.
  • the present inventors depending on the extraction method, contain a high concentration of usnic acid represented by the following formula in the extract of the genus Sarogase genus. I also found out.
  • Usnic acid is known as a hepatotoxic substance (for example, Frankos VH (January 2005), "NTP nomination for usnic acid and Usnea barbata herb", National Toxicology Program, http://ntp.niehs.nih.gov /ntp/htdocs/Chem_Background/ExSumPdf/UsnicAcid.pdf). Therefore, it is a problem to use a Salgogase lichen extract having a high content of difractic acid and a high content of usnic acid in a skin external preparation such as a whitening agent from the viewpoint of safety.
  • Salogase lichen is known to contain difractic acid and usnic acid (for example, Phytochemistry, 1987, vol. 26, No. 12, p. 3181-3185; Bull. Kochi. Gakuen College., 1992, vol.23, p.687 (9) -697 (19)).
  • the present inventors have studied the extraction conditions of Sarugagase lichen with reduced usnic acid content, which can be suitably used for skin external preparations such as whitening agents.
  • the concentration of usnic acid increases when an aqueous solution of alcohol with high alcohol content is used as the extraction solvent, while the concentration of usnic acid increases, while the aqueous solution of alcohol with a certain range of alcohol content is used as the extraction solvent.
  • the concentration of usnic acid can be reduced by extracting clothing.
  • an extract of Lichenaceae lichen extracted using an aqueous alcohol solution with a certain alcohol content range as an extraction solvent has an inhibitory effect on the increase in calcium (ion) concentration in melanocytes caused by the action of endothelin. .
  • the present inventors can use the Sargogase lichen extract or endothelin action inhibitor which can be used suitably for skin external preparations such as whitening agents and has a reduced content of toxic usnic acid.
  • Skin external preparations such as whitening agents
  • the concentration of difractic acid is significantly increased
  • the concentration of usnic acid, a hepatotoxic substance is significantly decreased.
  • the inventor has obtained knowledge that a Sargogase lichen extract or endothelin action inhibitor can be suitably used for a skin external preparation such as a whitening agent.
  • the present inventors can obtain a Sargogase lichen extract or endothelin action inhibitor that can be suitably used for a topical skin preparation such as a whitening agent and selectively reduces the content of toxic usnic acid.
  • the production method to be used was examined.
  • the concentration of usnic acid which is a hepatotoxic substance, is significant by performing low-temperature storage treatment and hydration treatment on the extract of Lichenaceae liquor prepared using an aqueous alcohol solution with an alcohol content exceeding 70% by volume as an extraction solvent. It was found that a Sargogase lichen extract or endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent is obtained.
  • the present invention has been completed based on these findings.
  • the present invention in particular, the Sargogus genus lichen extract obtained by the production method of the first embodiment of the present invention has a reduced content of usnic acid, which is a hepatotoxic substance. Therefore, the production method of the present invention, particularly the first embodiment of the present invention, produces an extract of Sarugagase lichen that can be suitably used for external skin preparations such as endothelin action inhibitors, melanin production inhibitors, and whitening agents. can do.
  • the present invention and in particular, the extract of Lichenaceae lichen and endothelin action inhibitor obtained by the production method of the second embodiment of the invention contains a high content of difractic acid, which is an active ingredient of endothelin action inhibitor, and is a hepatotoxic substance.
  • the content of usnic acid is reduced. Therefore, the production method of the present invention, particularly the second embodiment of the present invention, can produce a Salomagus lichen extract and an endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent.
  • the production method of the present invention can produce a Sargogase lichen extract or endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent.
  • the sorghum lichen is extracted using an aqueous alcohol solution having an alcohol content of 80% by volume or less. According to this method, an extract of Lichenaceae lichen having a low content of usnic acid, which is a hepatotoxic substance, can be obtained.
  • the extract of the genus Sarhogase obtained in this way is excellent in safety and should be suitably used for pharmaceuticals, quasi-drugs, cosmetics, etc. that can exhibit endothelin action inhibitory effect, melanin production inhibitory effect, whitening effect, etc. it can.
  • the term “usunic acid low-content Salogase lichen extract” refers to the extraction of Salogase lichen obtained using the extraction solvent defined in the first embodiment of the present invention when extraction is performed under the same conditions. It means that the usnic acid content in the product is less than the usnic acid content in the extract obtained using the extraction solvent having a higher alcohol content than the extraction solvent defined in the first embodiment of the present invention. In addition, the concentration of usnic acid in the extracts when the alcohol content is changed when the same solvent species is extracted under the same conditions is compared.
  • an alcohol having an alcohol content of 30% by volume to 80% by volume Extract Sarogase lichen using aqueous solution.
  • an extract of Salgocase genus lichen or endothelin action inhibitor containing difractic acid having an endothelin action inhibitory effect at a high concentration and a low content of usnic acid, which is a hepatotoxic substance is obtained.
  • the Saruogase lichen extract and endothelin action inhibitor thus obtained can be suitably used for pharmaceuticals, quasi-drugs, cosmetics and the like that can exert the whitening effect of difractic acid.
  • the extract of the genus Sarcogella having a high content of difructaic acid refers to the extraction of the genus Lichenaceus obtained using the extraction solvent defined in the second embodiment of the present invention when extraction is performed under the same conditions.
  • the content of difractic acid in the extract of Lichenacea lichen obtained using the extraction solvent defined in the second embodiment of the present invention is the second embodiment of the present invention.
  • the Lichenaceae lichen obtained using the extraction solvent defined in the second embodiment of the present invention is lower in the extract, the Lichenaceae lichen obtained using the extraction solvent defined in the second embodiment of the present invention
  • the extract is to be included in the “Dioglucate-rich and usnic acid-rich Salogase lichen extract” in the present specification.
  • the third embodiment of the present invention is meant to include all of the following fourth and fifth embodiments.
  • a sorghum lichen is extracted using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and hydration treatment.
  • a method for producing an extract that performs one or more selected treatments also referred to herein as “selective reduction treatment of usnic acid”
  • the residual ratio of usnic acid after the treatment is less than 50%
  • the fifth embodiment of the present invention is a method for extracting Sarcogum lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and comprising a low-temperature storage treatment and a hydration treatment for the obtained extract. It is a method for producing an extract containing a high content of difractic acid and a low content of usnic acid, or an endothelin action inhibitor, which performs at least one treatment selected from the group. According to this method, an extract of Saruogase lichen or endothelin action that contains difractic acid having an endothelin action inhibitory effect at a high concentration and a low content of usnic acid, which is a hepatotoxic substance, is obtained.
  • the Saruogase lichen extract and endothelin action inhibitor thus obtained can be suitably used for pharmaceuticals, quasi-drugs, cosmetics and the like that can exhibit a whitening effect.
  • the “usunic acid low-content Sarhogase lichen extract” refers to usnic acid contained in the Sarusose lichen extract extracted using the extraction solvent defined in the fourth and fifth embodiments of the present invention.
  • the proportion of the content of usnic acid in the extract of Lichenaceae lichen obtained by performing one or more treatments selected from the group consisting of low-temperature storage treatment and hydration treatment on the extract as compared with the content Is reduced.
  • the residual ratio of usnic acid after the treatment is less than 50% means that the usnic acid content in the extract after the selective reduction treatment of usnic acid is In consideration of dilution, when converted to the content before selective reduction treatment of usnic acid, the content of usnic acid in the extract after selective reduction treatment of usnic acid is before the selective reduction treatment of usnic acid. Is reduced to less than 50% of the usnic acid content in the extract.
  • the “difractate-rich Salogase lichen extract” is extracted using the extraction solvent defined in the fifth embodiment of the present invention when extraction is performed under the same conditions.
  • the content of difractic acid in the extract of Lichenaceae is higher than the content of difractic acid in the extract extracted using an extraction solvent having a lower alcohol content than the extraction solvent defined in the fifth embodiment of the present invention. This means that the concentration of difractor in the extract when the alcohol content is changed when extracted under the same conditions using the same solvent species is compared.
  • difractic acid may be present in the form of a salt.
  • the salt of difractic acid can be appropriately selected.
  • alkali metal salts such as lithium salt, sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, and alkylamine salts such as trimethylamine and triethylamine.
  • alkanolamine salts such as quaternary ammonium salts, triethanolamine, diethanolamine, and monoethanolamine, or amino acid salts such as lysine, histidine, and arginine.
  • alkali metal salts such as lithium salt, sodium salt and potassium salt
  • alkaline earth metal salts such as calcium salt and magnesium salt
  • alkylamine salts such as trimethylamine and triethylamine.
  • alkanolamine salts such as quaternary ammonium salts, triethanolamine, diethanolamine, and monoethanolamine, or amino acid salts such as lysine, histidine, and arginine.
  • the Lichenaceae lichen used in the present invention is a lichen belonging to the family Parmeliaceae.
  • naturally-occurring Salogase lichens may be collected and used, or commercially available products may be obtained and used.
  • Usnea territorial clothing for use in the present invention can be appropriately selected, Naga Usnea (Usnea longissima), Yokowa Usnea (Usnea diffracta Vain.), Fuji Usnea (Usnea trichodeoides), false Fuji Usnea (Usnea pseudomontis-fuji), Examples include Usnea pectinata .
  • the lichen genus lichen used in the present invention one or two or more selected from the group consisting of Nagasaru Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefujisaru Ogase and Kushinohasa Ogase are preferred, and the group consisting of Nagasara Ogase and Yokosawa Ogase 1 type or 2 types selected more are more preferable, and Nagasaru Ogase or Yokosawa Ogase is still more preferable.
  • any arbitrary part (the whole body, lichen body, temporary root, Iran, umbilical body, etc.) of the genus Ligoceae can be used, and a plurality of each part may be used in combination. .
  • standing culture cells fungal cells, algal cells, or an undifferentiated symbiotic cultured tissue in which both are mixed
  • standing culture cells fungal cells, algal cells, or an undifferentiated symbiotic cultured tissue in which both are mixed
  • the Sarhogase lichen used in the present invention may be in a naturally existing state, but in order to increase the extraction efficiency, it is preferable to use one subjected to treatment such as drying, freeze-drying, shredding, and pulverization.
  • an alcohol aqueous solution having an alcohol content of 80% by volume or less is used as an extraction solvent.
  • an alcohol aqueous solution having an alcohol content of 30% by volume to 80% by volume is used as the extraction solvent.
  • an aqueous alcohol solution having an alcohol content of more than 70% by volume is used as the extraction solvent.
  • aqueous alcohol solution used as the extraction solvent in the present invention examples include aqueous solutions of lower alcohols (preferably lower alcohols having 1 to 4 carbon atoms) such as methanol, ethanol, isopropyl alcohol, and butanol, 1,2-butanediol, 1 , 3-butanediol, 1,4-butanediol, 2,3-butanediol, aqueous solutions of polyhydric alcohols such as ethylene glycol, propylene glycol, and 1,3-butylene glycol.
  • these solvents may be used alone or in combination of two or more.
  • methanol, ethanol, isopropyl alcohol 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 2,3-butanediol, etc. It is preferable to use an aqueous solution, and it is more preferable to use an aqueous solution of methanol, ethanol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, or 2,3-butanediol. A 1,3-butanediol aqueous solution is more preferred.
  • an aqueous solution of lower alcohols such as methanol, ethanol, isopropyl alcohol, more preferably an aqueous solution of methanol or ethanol, and even more preferably an aqueous ethanol solution.
  • the alcohol content of the alcohol aqueous solution used as the extraction solvent is preferably 70% by volume or less, and more preferably 60% by volume or less.
  • the alcohol content of the aqueous alcohol solution used as the extraction solvent in the second embodiment of the present invention is preferably 40% by volume or more, preferably 70% by volume or less, and more preferably 60% by volume or less.
  • the alcohol content of the aqueous alcohol solution used as the extraction solvent in the third embodiment of the present invention is preferably more than 80% by volume, more preferably 90% by volume or more, and still more preferably 95% by volume or more.
  • the extraction temperature is preferably 0 ° C. or higher, more preferably 5 ° C. or higher, preferably 50 ° C. or lower, more preferably 40 ° C. or lower, and further preferably 20 ° C. or lower.
  • 0 to 50 ° C is preferable, 5 to 40 ° C is more preferable, and 5 to 20 ° C is further preferable.
  • the extraction time is preferably 1 hour or longer, more preferably 1 day or longer, preferably 20 days or shorter, more preferably 7 days or shorter. Alternatively, it is preferably 1 hour to 20 days, more preferably 1 to 7 days.
  • stirring may be performed during extraction, or homogenization treatment may be performed in a solvent.
  • the usage-amount of an extraction solvent can be set suitably, It is preferable that it is 1 time amount or more by mass ratio with respect to the Salogase lichen of an extraction raw material, It is more preferable that it is 5 times amount or more, 50 times amount Or less, more preferably 30 times or less. Alternatively, the amount is preferably 1 to 50 times, and more preferably 5 to 30 times.
  • the extract obtained by extraction with the solvent may be used as it is, but if necessary, further suitable separation means such as gel filtration, chromatography, A highly active fraction may be fractionated by precision distillation or the like.
  • the solvent of the obtained extract may be replaced with another solvent (for example, an aqueous alcohol solution).
  • the extract obtained by the production method of the present invention includes the various extracts thus obtained, a diluted solution thereof, a concentrated solution thereof, a purified product thereof or a dry powder thereof.
  • the extract obtained by extraction with the solvent may be used as it is for selective reduction of usnic acid, but if necessary, further suitable separation means such as a gel is used.
  • a highly active fraction may be fractionated by filtration, chromatography, precision distillation or the like.
  • the solvent of the obtained extract may be replaced with another solvent (for example, an aqueous alcohol solution).
  • the extract after solvent extraction used in the production method of the present invention includes the various extracts thus obtained, a diluted solution thereof, a concentrated solution thereof, a purified product thereof or a dry powder thereof.
  • the extract of Lichenaceae lichen obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent for the topical skin preparation
  • Any method can be selected as the usnic acid reduction treatment, and examples thereof include low-temperature storage treatment, water addition treatment, activated carbon treatment, hexane washing treatment, and heat treatment. In the present invention, these treatment steps may be used alone or in combination of two or more, and these treatment steps may be repeated.
  • an extract of Salogase lichen and an endothelin action inhibitor obtained using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is used as a skin external preparation.
  • a selective reduction treatment of usnic acid any method can be selected, and examples thereof include low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment, and heat treatment. In the present invention, these treatment steps may be used alone or in combination of two or more, and these treatment steps may be repeated. Even if the difractic acid content in the extract of Lichenaceae is reduced by the selective reduction treatment of usnic acid, the decrease rate of usnic acid content is sufficiently high compared to the decrease rate of difractic acid content. Good.
  • Salgogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment. Process more than seeds.
  • the selective reduction treatment of usnic acid may be used alone, or two or more may be used in combination, and these treatment steps may be repeated.
  • the order and number of times of the selective reduction of usnic acid are appropriately designed and changed in order to make the residual rate of usnic acid in the extract of Lichenaceae liquor less than 50%. be able to.
  • the low temperature storage as a (selective) reduction process of usnic acid will be described.
  • Usnic acid has high crystallinity, low solubility in aqueous solutions, and tends to precipitate at low temperatures. For this reason, when the Lichenaceae lichen extract extracted using the extraction solvent defined in the present invention is stored at a low temperature, usnic acid is hardly dissolved and crystals of usnic acid are precipitated. By removing the precipitated usnic acid by a conventional method, it becomes possible to (selectively) reduce the usnic acid from the Lichenaceae lichen extract.
  • the storage temperature of the extract of Lichenaceae is preferably ⁇ 10 ° C. or higher, more preferably ⁇ 5 ° C. or higher, and preferably 5 ° C.
  • 1st and 2nd embodiment of this invention as a preservation
  • the storage time of the extract of Lichenaceae liquor is preferably 1 hour or longer, more preferably 3 hours or longer, further preferably 6 hours or longer, preferably 40 days or shorter, 14 days
  • the following is more preferable, and 7 days or less is more preferable.
  • it is preferably 1 hour to 40 days, more preferably 3 hours to 14 days, and even more preferably 6 hours to 7 days.
  • the hydration treatment as a (selective) reduction treatment of usnic acid will be described.
  • Usnic acid has high crystallinity and low solubility in aqueous solutions. For this reason, when the Sarcophagus lichen extract extracted using the extraction solvent defined in the present invention is stored in a solution having a high water content, usnic acid is hardly dissolved and crystals of usnic acid are precipitated. By removing the precipitated usnic acid by a conventional method, it becomes possible to (selectively) reduce the usnic acid from the Lichenaceae lichen extract. In the first embodiment of the present invention, the amount of water added to the extract of Lichenaceae lichen can be appropriately set.
  • the amount of water added to the Saruogase lichen extract can be appropriately set within a range in which difractic acid is not lost due to precipitation or the like after addition. Specifically, it is preferable to add water so that the alcohol content after addition is 25% by volume or more, more preferably 30% by volume or more, and more preferably 45% by volume or less. It is preferable to add water, and it is more preferable to add water so as to be 40 volume% or less. Alternatively, it is preferable to add water so that the alcohol content after addition is 25 to 45% by volume, and it is more preferable to add 30% to 40% by volume.
  • the amount of water added to the Sarcophagus lichen extract it is possible to appropriately set the amount of water added to the Sarcophagus lichen extract, and it is preferable to add water so that the alcohol content after addition is 25% by volume or more, and 30 volumes. It is more preferable to add water so as to be not less than 50%, preferably not more than 50% by volume, and more preferably not more than 45% by volume. Alternatively, it is preferable to add water so that the alcohol content after addition is 25 to 50% by volume, and it is more preferable to add 30% to 45% by volume.
  • the activated carbon treatment as a (selective) reduction treatment of usnic acid will be described.
  • usnic acid is particularly easily adsorbed on activated carbon.
  • difractic acid and usnic acid differ in molecular size and compound physical properties (polarity). Because of these differences, usnic acid is more easily adsorbed on activated carbon than difractic acid. Therefore, by applying the activated carbon treatment to the extract of the genus Sarhogase obtained by the method of the present invention, usnic acid in the extract can be (selectively) removed.
  • the amount of the activated carbon can be appropriately set, and is preferably 1% by weight or more, more preferably 10% by weight or more, still more preferably 50% by weight or more, and 200% by weight with respect to the solid content of the extract of Lichenaceae.
  • the following is preferable, and 150% by weight or less is more preferable.
  • it is preferably 1 to 200% by weight, more preferably 10 to 200% by weight, and further preferably 50 to 150% by weight.
  • the treatment time can also be set as appropriate, preferably 10 minutes or longer, more preferably 30 minutes or longer, preferably 24 hours or shorter, more preferably 1 hour or shorter. Alternatively, 10 minutes to 24 hours are preferable, and 30 minutes to 1 hour are more preferable.
  • the Saruogase lichen extract and activated carbon may be mixed and stirred.
  • the activated carbon may be a normal one, and a commercially available one can be used.
  • a hexane washing process as a (selective) reduction process of usnic acid will be described.
  • usnic acid is particularly easily dissolved in hexane.
  • difractic acid is difficult to dissolve in hexane, whereas usnic acid is easily dissolved in hexane.
  • the usanic acid lichen extract is subjected to hexane washing treatment and the hexane layer is separated to remove (selectively) usnic acid in the sorghum lichen extract.
  • the method of washing with hexane can be selected as appropriate.
  • the hexane washing lichen extract and hexane, and optionally mixed with an alcohol solution such as ethanol, are separated into two layers, and the layer containing the lichen extract is usually added. It may be recovered by the law. Further, the hexane cleaning process may be repeated.
  • the heat treatment as a (selective) reduction treatment of usnic acid will be described. It is thought that heat treatment is performed on the extract of Lichenaceae liquor, the other components in the lichen extract of Lilium extract react with usnic acid, decomposition of usnic acid occurs, and the content of usnic acid decreases. It is done.
  • the heating temperature of the extract of Lichenaceae is preferably 50 ° C. or higher, more preferably 60 ° C. or higher, preferably 100 ° C. or lower, more preferably 90 ° C. or lower, and further preferably 80 ° C. or lower.
  • the heating time of the Saruogase lichen extract can be appropriately set depending on the heating temperature, preferably 1 hour or more, more preferably 6 hours or more, preferably 7 days or less, and more preferably 3 days or less. Alternatively, it is preferably 1 hour to 7 days, more preferably 6 hours to 3 days.
  • a low-temperature storage treatment, a water addition treatment, an activated carbon treatment, and a hexane washing is performed on a Salgogase lichen extract obtained using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent. It is preferable to further reduce usnic acid by one or more treatments selected from the group consisting of treatment and heat treatment, and more preferable methods for reducing usnic acid include the following. (1) The extracted Saruogase lichen extract is further subjected to a hydration treatment and a low-temperature storage treatment. (2) A low-temperature storage treatment is further performed on the extracted Sarhogase lichen extract.
  • a low temperature preservation process, a hydration process, and a low temperature preservation process are further performed in this order with respect to the extracted Sarhogase lichen extract.
  • a hydrotreating process and a low-temperature storage process are performed in this order on the extracted Lichenaceae lichen extract.
  • a hydrous treatment, a low-temperature storage treatment, and an activated carbon treatment are performed in this order on the extracted Saruogase lichen extract.
  • a hydrous treatment, a low-temperature storage treatment, and a hexane washing treatment are performed in this order on the extracted Sarhogase lichen extract.
  • the low-temperature storage treatment and the activated carbon treatment are further performed in this order on the extracted Lichenaceae lichen extract.
  • the low-temperature storage treatment and the hexane washing treatment are further performed in this order on the extracted Sarhogase lichen extract.
  • the extracted Saruogase lichen extract is further subjected to a water treatment, a low-temperature storage treatment, and a heat treatment in this order.
  • the absolute amount of usnic acid can be appropriately set, and the usnic acid content is preferably 150 ppm or less, preferably 120 ppm or less. More preferably it is.
  • a low-temperature storage treatment a water addition treatment, activated carbon is applied to a Salugatace lichen extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume as an extraction solvent.
  • a Salugatace lichen extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume as an extraction solvent.
  • the extracted Saruogase lichen extract is further subjected to a hydration treatment and a low-temperature storage treatment.
  • a low-temperature storage treatment is further performed on the extracted Sarhogase lichen extract.
  • a low temperature preservation process, a hydration process, and a low temperature preservation process are further performed in this order with respect to the extracted Sarhogase lichen extract.
  • a hydrotreating process and a low-temperature storage process are performed in this order on the extracted Lichenaceae lichen extract.
  • a hydrous treatment, a low-temperature storage treatment, and an activated carbon treatment are performed in this order on the extracted Saruogase lichen extract.
  • a hydrous treatment, a low-temperature storage treatment, and a hexane washing treatment are performed in this order on the extracted Sarhogase lichen extract.
  • the low-temperature storage treatment and the activated carbon treatment are further performed in this order on the extracted Lichenaceae lichen extract.
  • the low-temperature storage treatment and the hexane washing treatment are further performed in this order on the extracted Sarhogase lichen extract.
  • the extracted Saruogase lichen extract is further subjected to a water treatment, a low-temperature storage treatment, and a heat treatment in this order.
  • the content ratio of difractic acid relative to usnic acid can be set as appropriate, and the obtained Sarcoses lichen extract obtained.
  • the usnic acid content in the endothelin action inhibitor is set to 1 on a mass basis
  • the difractic acid content is preferably 3 or more, more preferably 5 or more.
  • the absolute amount of usnic acid can be set as appropriate, and the usnic acid content is preferably 150 ppm or less, and more preferably 120 ppm or less.
  • the content of active ingredients such as endothelin action inhibitor, melanin production inhibitor, whitening agent, etc. contained in the extract of Lichenaceae by selective reduction of usnic acid is reduced.
  • the residual ratio of usnic acid is sufficiently reduced.
  • the difractic acid content in the extract of Lichenaceae is reduced by the selective reduction treatment of usnic acid, the residual ratio of usnic acid relative to the residual ratio of difractic acid Should be low enough.
  • the extraction using the aqueous alcohol solution, the hydration treatment, and the low-temperature storage treatment it is preferable to perform the extraction using the aqueous alcohol solution, the hydration treatment, and the low-temperature storage treatment in this order. It is also preferable to perform the extraction using the aqueous alcohol solution, the low-temperature storage treatment, the hydration treatment, and the low-temperature storage treatment in this order.
  • the content ratio of difractic acid to usnic acid can be appropriately set, and the extract of Lizoaceae lichen or endothelin action inhibitor obtained
  • the content of usnic acid is 1 on the mass basis
  • the content of difractic acid is preferably 2 or more, and more preferably 10 or more.
  • the absolute amount of usnic acid can be set as appropriate, and the usnic acid content is preferably 300 ppm or less, more preferably 200 ppm or less.
  • the aqueous solution is used to treat the sorghum lichen before the extraction of the sorghum lichen using the aqueous alcohol solution, and the aqueous solvent is removed. Extraction using the aqueous alcohol solution may be performed on the residue of the Lichenaceae lichen obtained in this way.
  • a water-soluble component contained in the sorghum lichen can be removed by treating the sorghum lichen with an aqueous solvent, and the extract of the sorghum lichen extract or endothelin action inhibitor with less impurities, or difractic acid concentration with less impurities Can be produced, particularly a Sargogase lichen extract or endothelin action inhibitor.
  • aqueous solvent a solvent containing a water content sufficient to remove water-soluble components contained in the genus Lisaceae can be appropriately selected.
  • Water, an alcohol aqueous solution for example, an aqueous methanol solution, an aqueous ethanol solution
  • Propanol aqueous solution isopropanol aqueous solution
  • water or an alcohol aqueous solution having an alcohol content of 10% by volume or less, preferably 5% by volume or less, more preferably 1% by volume or less is preferable.
  • Arbitrary methods can be selected as a method for treating S. aureus lichen using the above-mentioned aqueous solvent, and a method of immersing S. aureus lichen in an aqueous solvent, a method of washing a sarcoma lichen with an aqueous solvent, etc. .
  • a method of immersing S. aureus lichen in an aqueous solvent in order to increase the efficiency of removing water-soluble components contained in the sorghum lichen, stirring may be performed during the treatment or homogenization treatment may be performed in the solvent.
  • the treatment conditions for Sargogase lichen using an aqueous solvent can be appropriately set depending on the solvent used.
  • the treatment temperature is preferably 0 ° C. or higher, more preferably 5 ° C. or higher, preferably 50 ° C. or lower, and more preferably 30 ° C. or lower.
  • 0 to 50 ° C. is preferable, and 0 to 30 ° C. is more preferable.
  • the treatment time is preferably 5 hours or longer, more preferably 1 day or longer, preferably 10 days or shorter, and more preferably 7 days or shorter.
  • the S. aeruginosa lichen is treated with a hydrophobic organic solvent such as hexane. You may perform extraction using the said aqueous alcohol solution with respect to the residue of Salgogase lichen obtained by removing hydrophobic substances, such as an acid.
  • Hydrophobic components contained in Sorghum lichen can be removed by treating Sorghum lichen using a hydrophobic organic solvent, and the extract of Lactobacillus lichen extract or endothelin action inhibitor with little impurities, or difractor with few impurities It is possible to produce an extract of Saruogase lichen or an endothelin action inhibitor having a particularly high acid concentration.
  • hydrophobic organic solvent a solvent capable of removing the hydrophobic component contained in the garlic lichen can be appropriately selected, and n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane. , Cycloheptane, octane, and other linear alkanes that are liquid at room temperature and pressure, and n-hexane is preferred.
  • the processing method of the Sarcophagus lichen using the said hydrophobic organic solvent it can choose suitably, The method of immersing the Sarcophagus lichen in the hydrophobic organic solvent, The method of wash
  • stirring may be performed during the treatment or a homogenization treatment may be performed in the solvent. .
  • the processing conditions for the Sarcophagus lichen using a hydrophobic organic solvent can be appropriately set depending on the solvent used.
  • the treatment temperature is preferably 0 ° C. or higher, more preferably 5 ° C. or higher, preferably 50 ° C. or lower, and more preferably 30 ° C. or lower.
  • 0 to 50 ° C. is preferable, and 0 to 30 ° C. is more preferable.
  • the treatment time is preferably 5 hours or longer, more preferably 1 day or longer, preferably 10 days or shorter, and more preferably 7 days or shorter.
  • Nagasagaogase extract having a low content of usnic acid which is a hepatotoxic substance
  • Nagasagaogase extract has a melanin production inhibitory action and an endothelin action inhibitory action shown in the examples described later. Therefore, the Ususic acid low-content Salogase lichen extract obtained by the production method of the first embodiment of the present invention is It is highly safe and can be suitably used for an external preparation for skin that exhibits a whitening effect, such as an endothelin inhibitor and a melanin production inhibitor.
  • the extraction process WHEREIN The Salomasae lichen extract and the endothelin action inhibitor which are high in the content of difractic acid and low in the content of usnic acid are obtained. be able to. Therefore, since difractic acid can suppress the production of melanin by suppressing the action of endothelin on melanocytes, the Saruogase lichen extract and endothelin action inhibitor obtained by the production method of the second embodiment of the present invention are excellent. It has an endothelin action inhibitory effect and a whitening effect.
  • the extract of the genus Sarugagase liquor which fully reduced the content of usnic acid can be obtained.
  • an extract of Salgocase lichen and endothelin action inhibitor having a high difractic acid content and a low usnic acid content.
  • the Saruogase genus lichen extract has a melanin production inhibitory effect and an endothelin action inhibitory effect shown in Examples described later.
  • Difractic acid also has an endothelin action inhibitory effect.
  • the Sarcophagus lichen extract and endothelin action inhibitor obtained by the production method of the third embodiment of the present invention can suppress the production of melanin by inhibiting the action of endothelin on melanocytes
  • the endothelin inhibitor, melanin can be used for an external preparation for skin that exhibits a whitening effect, such as a production inhibitor.
  • whitening refers to suppressing melanin pigment formation and pigmentation, returning to an original transparent skin color without excess melanin, or pigmentation such as skin blackening or stain / sobacus. It means to prevent and suppress.
  • the sarcoma lichen extract obtained by the production method of the second and third embodiments of the present invention and the endothelin action inhibitor have a low content of usnic acid, which is a hepatotoxic substance.
  • endothelin action inhibitor has high safety and can be suitably used as a skin external preparation such as a whitening agent.
  • the Saruogase lichen extract obtained by the present invention may be used as an endothelin inhibitor or a whitening agent as it is.
  • the Lichenaceae lichen extract obtained by the present invention may be used by mixing with a suitable liquid or solid excipient or extender such as titanium oxide, calcium carbonate, distilled water, lactose, starch and the like.
  • a suitable liquid or solid excipient or extender such as titanium oxide, calcium carbonate, distilled water, lactose, starch and the like.
  • other endothelin action inhibitors, other whitening agents, moisturizers, antioxidants, UV absorbers, surfactants, thickeners, colorants, oily substances, polymer compounds, preservatives, powders , Pigments, fragrances, emulsion stabilizers, pH adjusters, and the like may be used in admixture with components used in ordinary whitening agents.
  • difractic acid when using the Saruogase genus lichen extract obtained by the 4th embodiment of this invention for an endothelin action inhibitor or a whitening agent, it is good also considering difractic acid as an active ingredient and making another ingredient into an active ingredient.
  • difractic acid when using the Sarhogase lichen extract obtained by the fifth embodiment of the present invention as an endothelin action inhibitor or whitening agent, only difractic acid may be used as an active ingredient, or in combination with difractic acid and other ingredients. It may be an active ingredient.
  • the endothelin action inhibitor obtained by the method of the present invention and the whitening agent of the present invention can be applied to the skin and the like as skin cosmetics, external medicines, quasi-drugs, etc. It can take a wide variety of forms such as a solubilization system, an emulsification system, a powder system, a gel system, an ointment system, a cream, a water-oil two-layer system, and a water-oil-powder three-layer system. Examples thereof include face wash, lotion, milky lotion, cream, lotion, gel, essence (beauty serum), pack, mask, foundation, ointment, sheet-like product and the like.
  • the content of the Sargogus genus lichen extract in the endothelin action inhibitor obtained by the method of the present invention and the whitening agent of the present invention can be appropriately set.
  • it can be set as appropriate depending on the concentration of an active ingredient such as difractic acid in the extract of Lichenaceae lichen.
  • the amount of endothelin action inhibitor obtained by the method of the present invention and the amount of whitening agent of the present invention can be appropriately set and varies depending on the content of the active ingredient.
  • the skin surface 0.1 ⁇ g or more per 1 cm 2 is preferable, and 5 ⁇ g or less is preferable.
  • 0.1 to 5 ⁇ g per 1 cm 2 of the skin surface is preferable.
  • 0.1 ⁇ g or more per 1 cm 2 of skin surface is preferable, and 10 ⁇ g or less is preferable.
  • 0.1 to 10 ⁇ g per 1 cm 2 of the skin surface is preferable.
  • the present invention further discloses the following extract production method, endothelin action inhibitor, whitening agent, method and use.
  • a Salmonella lichen extract containing a low amount of usnic acid which is extracted using an aqueous alcohol solution having an alcohol content of 80% by volume or less, preferably 70% by volume or less, more preferably 60% by volume or less. Manufacturing method.
  • aqueous alcohol solution having an alcohol content of 30% by volume or more preferably 40% by volume or more and 80% by volume or less, preferably 70% by volume or less, more preferably 60% by volume or less.
  • ⁇ 3> The method according to ⁇ 1> or ⁇ 2>, wherein the alcohol aqueous solution is an ethanol aqueous solution or a 1,3-butanediol aqueous solution.
  • ⁇ 4> At least one selected from the group consisting of Nagasaga Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefu Gisal Ogase and Kushinohasaru Ogase, preferably at least one selected from the group consisting of Nagasaro Ogase and Yokosawa Ogase
  • ⁇ 5> selective) reduction of usnic acid by one or more treatments selected from the group consisting of low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment, and heat treatment on the extracted Lichenaceae lichen extract
  • ⁇ 6> The method according to ⁇ 5> above, wherein the extracted extract of Sorghum lichen is subjected to water treatment and low-temperature storage treatment to perform (selective) reduction of the usnic acid.
  • ⁇ 7> The production method according to ⁇ 5> above, wherein the extracted extract of the genus Sarhogase is subjected to a low-temperature storage treatment to perform a (selective) reduction treatment of the usnic acid.
  • ⁇ 8> The production according to ⁇ 5>, wherein the extracted extract of the genus Sarhogase is subjected to a low-temperature storage treatment, a hydration treatment, and a low-temperature storage treatment in this order to perform (selective) reduction of the usnic acid.
  • Aqueous solvent preferably water or an alcohol content of 10% by volume or less (preferably 5% by volume or less, more preferably 1% by volume or less)) (preferably methanol aqueous solution, ethanol aqueous solution, propanol aqueous solution) Or the aqueous solution of isopropanol)), and the extracted aqueous liquor is extracted with the aqueous alcohol solution, according to any one of the above items ⁇ 1> to ⁇ 14> Production method.
  • a hydrophobic organic solvent preferably at least one hydrophobic organic solvent selected from the group consisting of n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane, cycloheptane and octane, more preferably n
  • n-pentane cyclopentane
  • n-hexane cyclohexane
  • n-heptane cycloheptane and octane
  • n preferably n
  • the sarcoma lichen is treated with -hexane
  • the treated sarcoma lichen is extracted with the aqueous alcohol solution.
  • Salogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, preferably more than 80% by volume, more preferably 90% by volume or more, and even more preferably 95% by volume or more.
  • a method for producing an extract wherein the obtained extract is subjected to at least one treatment selected from the group consisting of a low-temperature storage treatment and a hydrolysis treatment, wherein the residual ratio of usnic acid after the treatment is less than 50%
  • Salogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, preferably more than 80% by volume, more preferably 90% by volume or more, and even more preferably 95% by volume or more.
  • ⁇ 20> The method according to ⁇ 18> or ⁇ 19>, wherein the aqueous alcohol solution is an aqueous ethanol solution.
  • ⁇ 21> At least one selected from the group consisting of Nagasaro-Ogase, Yokosawa-Ogase, Fujisaru-Ogase, Nisefu-jisaru-ogase and Kushinohasaru-ogase, preferably at least one selected from the group consisting of Nagasaro-ogase and Yokosawa-o-gase
  • the production method according to any one of ⁇ 18> to ⁇ 20>, wherein ⁇ 22> The production method according to any one of ⁇ 18> to ⁇ 21>, wherein the extraction using the aqueous alcohol solution, the hydrolysis treatment, and the low-temperature storage treatment are performed in this order.
  • ⁇ 23> The method according to any one of ⁇ 18> to ⁇ 22>, wherein the extraction using the aqueous alcohol solution, the low-temperature storage treatment, the hydration treatment, and the low-temperature storage treatment are performed in this order.
  • ⁇ 24> Any one of the above ⁇ 19> to ⁇ 23>, wherein when the usnic acid content in the obtained extract is 1 on a mass basis, the difractic acid content is 2 or more, preferably 10 or more The manufacturing method of 1 item
  • An endothelin action inhibitor comprising, as an active ingredient, an extract of the genus Sarhogase obtained by the production method according to any one of ⁇ 1> to ⁇ 16>.
  • a whitening agent comprising, as an active ingredient, an extract of a Lichenaceae lichen obtained by the production method according to any one of ⁇ 1> to ⁇ 16>.
  • Sarhogase lichen having an alcohol content of 30% by volume or more, preferably 40% by volume or more and 80% by volume or less, preferably 70% by volume or less, more preferably 60% by volume or less.
  • ⁇ 28> The production method according to ⁇ 27>, wherein the alcohol aqueous solution is an ethanol aqueous solution or a 1,3-butanediol aqueous solution.
  • ⁇ 29> At least one selected from the group consisting of Nagasaga Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefu Gisal Ogase, and Kushinohasaru Ogase, preferably at least one selected from the group consisting of Nagasaro Ogase and Yokosawa Ogase
  • the method according to ⁇ 27> or ⁇ 28>, wherein ⁇ 30> Extracted from the genus Lichenaceae liquor, extracted from the genus Lichenaceae liquor by one or more treatments selected from the group consisting of low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment and heat treatment The production method according to any one of ⁇ 27> to ⁇ 29>, wherein the selective reduction treatment of usnic acid
  • a low-temperature preservation treatment, a hydrolyzing preservation treatment, and a low-temperature preservation treatment are performed in this order on the extracted Saruogase lichen extract in this order to perform a selective reduction treatment of usnic acid contained in the Saruogase lichen extract, ⁇ 30> The production method according to item.
  • the extract of Salgogase lichen is subjected to water treatment, low-temperature storage treatment and activated carbon treatment in this order to perform selective reduction treatment of usnic acid contained in the Salgogase lichen extract, ⁇ 30 > The manufacturing method of description.
  • the extract of Salgogase lichen is subjected to water treatment, low-temperature storage treatment, and hexane washing treatment in this order to perform a selective reduction treatment of usnic acid contained in the Salgogase lichen extract. Item 30>.
  • the extract of Salgogase lichen is subjected to water treatment, low-temperature storage treatment, and heat treatment in this order to perform a selective reduction treatment of usnic acid contained in the Salgogase lichen extract, ⁇ 30 > The manufacturing method of description.
  • aqueous alcohol solution preferably water or an alcohol content of 10% by volume or less (preferably 5% by volume or less, more preferably 1% by volume or less)) (preferably methanol aqueous solution, ethanol aqueous solution, propanol aqueous solution) Or the aqueous solution of isopropanol)
  • the extracted aqueous liquor is extracted with the aqueous alcohol solution, according to any one of the above items ⁇ 27> to ⁇ 39> Production method.
  • a hydrophobic organic solvent preferably at least one hydrophobic organic solvent selected from the group consisting of n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane, cycloheptane and octane, more preferably n
  • a hydrophobic organic solvent preferably at least one hydrophobic organic solvent selected from the group consisting of n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane, cycloheptane and octane, more preferably n
  • sarcoma lichen is treated using (-hexane)
  • the treated sarcophagus lichen is extracted using the aqueous alcohol solution.
  • the content of usnic acid in the obtained endothelin inhibitor is set to 1 on a mass basis
  • the content of difractic acid is 3 or more, preferably 5 or more, ⁇ 27> to ⁇ 41>
  • a whitening agent comprising, as an active ingredient, a Salugose lichen extract or endothelin action inhibitor obtained by the production method according to any one of ⁇ 1> to ⁇ 17> and ⁇ 27> to ⁇ 42>.
  • Salogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, preferably more than 80% by volume, more preferably 90% by volume or more, and even more preferably 95% by volume or more.
  • the manufacturing method of the endothelin action inhibitor which performs 1 or more types of processes chosen from the group which consists of a low-temperature preservation
  • ⁇ 45> The method according to ⁇ 44>, wherein the aqueous alcohol solution is an aqueous ethanol solution.
  • ⁇ 46> At least one selected from the group consisting of Nagasaga Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefujisaru Ogase, and Kushinohasa Ogase, preferably at least one selected from the group consisting of Nagasaro Ogase and Yokosawa Ogase
  • the method according to ⁇ 44> or ⁇ 45>, wherein ⁇ 47> The production method according to any one of ⁇ 44> to ⁇ 46>, wherein the extraction using the aqueous alcohol solution, the hydrolysis treatment, and the low-temperature storage treatment are performed in this order.
  • ⁇ 48> The method according to any one of ⁇ 44> to ⁇ 46>, wherein the extraction using the aqueous alcohol solution, the low-temperature storage treatment, the hydration treatment, and the low-temperature storage treatment are performed in this order.
  • the difractic acid content is 2 or more, preferably 10 or more, ⁇ 44> to ⁇ 48>.
  • a whitening agent comprising, as an active ingredient, an extract of Salogase lichen or an endothelin action inhibitor obtained by the production method according to any one of ⁇ 18> to ⁇ 24> and ⁇ 44> to ⁇ 48> .
  • ⁇ 51> Use of an extract of Ligocephalus lichen obtained by the production method according to any one of ⁇ 1> to ⁇ 24> as an endothelin action inhibitor or a whitening agent.
  • ⁇ 52> Use of an extract of Salogase lichen obtained by the production method according to any one of ⁇ 1> to ⁇ 24> for the production of an endothelin action inhibitor or a whitening agent.
  • ⁇ 53> A method of using the Salgogase lichen extract obtained by the production method according to any one of ⁇ 1> to ⁇ 24> as an endothelin action inhibitor or a whitening agent.
  • ⁇ 54> A method for suppressing endothelin action, which uses the Salgaraceae lichen extract obtained by the production method according to any one of ⁇ 1> to ⁇ 24>.
  • ⁇ 55> A whitening method using the Salgogase lichen extract obtained by the production method according to any one of ⁇ 1> to ⁇ 24>.
  • ⁇ 56> The use or method according to any one of ⁇ 51> to ⁇ 55>, wherein the increase in calcium ion concentration in melanocytes is suppressed to suppress endothelin action.
  • ⁇ 57> The use or method according to any one of the above items ⁇ 51> to ⁇ 56>, wherein the extract of Lichenaceae lichen is applied in the form of a cosmetic.
  • ⁇ 58> Use of an endothelin action inhibitor obtained by the production method according to any one of ⁇ 27> to ⁇ 42> and ⁇ 44> to ⁇ 49> as a whitening agent.
  • ⁇ 59> Use of an endothelin action inhibitor obtained by the production method according to any one of ⁇ 27> to ⁇ 42> and ⁇ 44> to ⁇ 49> for the production of a whitening agent.
  • ⁇ 60> A method of using the endothelin action inhibitor obtained by the production method of any one of ⁇ 27> to ⁇ 42> and ⁇ 44> to ⁇ 49> as a whitening agent.
  • ⁇ 61> A whitening method using an endothelin action inhibitor obtained by the production method according to any one of ⁇ 27> to ⁇ 42> and ⁇ 44> to ⁇ 49>.
  • ⁇ 62> The use or method according to any one of ⁇ 58> to ⁇ 61>, wherein the increase in calcium ion concentration in melanocytes is suppressed to suppress endothelin action.
  • ⁇ 63> The use or method according to any one of ⁇ 58> to ⁇ 62>, wherein the endothelin action inhibitor is applied in the form of a cosmetic.
  • German chamomile extract 40 g of flower portion of German chamomile (Japanese name: chamomile, obtained from Shinwa Bussan Co., Ltd.) and 400 mL of 50% ethanol-containing aqueous solution were mixed and extracted after 14 days at room temperature and filtered German chamomile extract (221 mL) was obtained (evaporation residue 2.63%).
  • Endothelin action inhibitory effect About Nagasaru Ogase extract, Yokosawa Ogase extract, German chamomile extract and difractic acid, the endothelin action inhibitory effect was verified using the following evaluation system.
  • Endothelin is known to act on melanocytes to increase calcium concentration and increase melanin production (Yada Y. et al., J. Biol. Chem., 1991, vol. 266, p. 18352). -18357; Imokawa G. et al., J. Biol. Chem., 1992, vol. 267, p. 24675-24680, etc.), substances that suppress the action of endothelin are useful as whitening ingredients.
  • NHEMs Normal human neonatal epidermis-derived melanocytes
  • NHEMs Normal human neonatal epidermis-derived melanocytes
  • HMGS PMA growth additive
  • a 50% by volume ethanol aqueous solution or dimethyl sulfoxide added was used as a control. The results are shown in Table 2.
  • German chamomile extract is known to suppress the increase in Ca concentration of melanocytes by the action of endothelin and suppress melanin production (see, for example, Pigment Cell Res., 1997, vol. 10, p. 218-228). ).
  • Table 2 in the system to which the German chamomile extract was added, a concentration-dependent calcium concentration increase inhibitory action was confirmed.
  • an increase in calcium concentration was suppressed by 20% or more, and an excellent endothelin inhibitory action was confirmed.
  • Example 1-1 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, 207.35 g of Nagasaru Ogase extract (evaporation residue 0.202 (w / w v)%).
  • Example 1-2 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed and extracted at 30 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 206.59 g (evaporation residue 0.266 (w / v)%).
  • Example 1-3 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed, extracted at 40 ° C. for 7 days, filtered, and Nagasal Ogase extract 208.26 g (evaporation residue 0.280 (w / w v)%).
  • Example 1-4 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 20 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.49 g (evaporation residue 0.172 (w / v)%).
  • Example 1-5 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 30 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 199.90 g (evaporation residue 0.146 (w / v)%).
  • Example 1-6 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 40 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasal Ogase extract 200.70 g (evaporation residue 0.190 (w / v)%).
  • Example 1-7 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 5 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.67 g (evaporation residue 0.178 (w / v)%).
  • Example 1-8 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 15 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 202.57 g (evaporation residue 0.192 (w / v)%).
  • Example 1-9 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.69 g (evaporation residue 0.298 (w / v)%).
  • Example 1-10 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 30 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 200.54 g (evaporation residue 0.258 (w / v)%).
  • Example 1-11 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.49 g (evaporation residue 0.316 (w / v)%).
  • Example 1-12 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 60 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 198.69 g (evaporation residue 0.242 (w / v)%).
  • Example 1-13 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 70 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, 195.1 g of Nagasal Ogase extract (evaporation residue 0.232 (w / v)%).
  • Example 1-14 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 80 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.6 g (evaporation residue 0.216 (w / v)%).
  • Measuring instruments HPLC: Agilent Technologies Open LAB CDS LC02-1260, Autosampler G1329B 1260 ALS, Column oven G1316A 1260 TCC, Detector G1315D 1260 DAD VL, Pump G1312B 1260 Bin pump, G1379B 1260 ⁇ -Degasser ⁇ HPLC analysis conditions> Flow rate: 0.75 mL / min Column used: Inertsil ODS-3 5 ⁇ m (GL Science), 3.0 ID ⁇ 150 mm Detection wavelength: 254 nm Column temperature: 40 ° C Mobile phase: A: 0.1% trifluoroacetic acid-containing aqueous solution, B: 0.1% trifluoroacetic acid-containing methanol 0 to 10 minutes: 70% B 10-15 minutes: 70% B ⁇ 100% B 15-20 minutes: 100% B The results are shown in Table 3.
  • a Salusose lichen extract containing a low amount of usnic acid can be produced by extracting the Ligosa lichen using an aqueous alcohol solution having an alcohol content of 80% by volume or less. Since usnic acid has hepatotoxicity, according to the first embodiment of the present invention, an extract of Sargogase lichen that is excellent in safety and can be suitably used for an external skin preparation such as an endothelin action inhibitor is produced. be able to.
  • Example 1-14 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasaga Ogase extract 858.5 g (evaporation residue 0.374 (w / v)%). The obtained extract was stored at ⁇ 5 ° C. for 12 days and then filtered to obtain 833.4 g of Nagasaroogase extract (low evaporation residue 0.360 (w / v)%) after the low-temperature storage treatment. The Usunic acid content was measured by the same method as above for the Nagasaga Ogase extract before and after the low-temperature storage treatment. The results are shown in Table 4.
  • the Ussanoic acid content is further reduced by performing low-temperature storage treatment on the Nagasaruogase extract obtained using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent. can do.
  • Example 1-16 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasal Ogase extract 852.8 g (evaporation residue 0.268 (w / w v)%). The obtained extract was stored at ⁇ 5 ° C. for 13 days and filtered to obtain 837.2 g (evaporation residue 0.256 (w / v)%) of the Nagasaga Ogase extract before treatment. Next, 10 mL of Nagasaga Ogase extract pretreatment product and 2.5 mL of distilled water were mixed and diluted so that the volume of ethanol was 40%. .
  • the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 4 days and then filtered, and the Nagasal Ogase extract after the hydrous / low temperature preservation treatment was 11.1 g (evaporation residue 0.130 (w / v)%).
  • Got. Usasic acid content was measured by the same method as above for the Nagasaga Ogase extract before treatment, the Nagasaga Ogase extract after the hydrotreatment, and the Nagasal Ogase extract after the hydrothermal treatment at low temperature. The results are shown in Table 5.
  • the theoretical value of the usnic acid content contained in the Nagasaru Ogase extract after the hydrotreatment is also shown in Table 5.
  • Example 1-17 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 1 L of 50% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 7 days, filtered, and 838.5 g (Evaporation residue) Min 0.218 (w / v)%). Next, 800 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 200 mL of distilled water are mixed and diluted so that the volume of ethanol becomes 40%. Obtained. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C.
  • Nagasaga Ogase extract is obtained by subjecting Nagasaga Ogase extract obtained by using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent to water treatment and low-temperature storage treatment. Can further reduce the usnic acid content.
  • Example 1-18 5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. Then, add 0.65 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (10% by weight based on the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 10 wt% -treated product was obtained.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment.
  • the acid content was measured (evaporation residue 0.08 (w / v)%)). The results are shown in Table 7.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment.
  • the acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
  • activated carbon trade name: Shirahige M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment.
  • the acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
  • Example 1-21 5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. Then, 9.75 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (150% by weight with respect to the solid content in the extract) is added, and the mixture is stirred at room temperature for 45 minutes and then filtered. Thus, a product after treatment with 150 wt% of activated carbon was obtained.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment.
  • the acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
  • Example 1-22 5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. 13.0 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (200% by weight with respect to the solid content in the extract) is added, stirred for 45 minutes at room temperature, and then filtered. Thus, a product after treatment with 200 wt% of activated carbon was obtained.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment.
  • the acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
  • Nagasagaogase extract obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and activated carbon treatment.
  • the usnic acid content contained in the Saruogase extract can be further reduced.
  • Example 1-23 50 mL of hexane and 5 mL of 99.5 vol% ethanol aqueous solution were added to 50 mL of Nagasaroogase extract (extract before hexane washing) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment, and a separating funnel was used. After separating into two layers, the lower layer was recovered (hexane washed once product), and the usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 8.
  • Example 1-24 To 50 mL of Nagasaga Ogase extract obtained in Example 1-23, hexane 50 mL and 99.5% by volume aqueous ethanol solution 5 mL were added again, and after separating into two layers using a separatory funnel, the lower layer was recovered (hexane The usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 8.
  • Example 1-25 Add 45 mL of hexane again to 45 mL of Nagasaga Ogase extract obtained in Example 1-24, separate into two layers using a separatory funnel, and recover the lower layer (hexane washed three times), as above
  • the usnic acid content was measured by HPLC according to the above method. The results are shown in Table 8.
  • the Nagasaruogase extract obtained using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and hexane washing treatment.
  • the usnic acid content contained in the Nagasaru Ogase extract can be further reduced.
  • Example 1-26 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% aqueous ethanol solution 1 L were mixed and extracted at 20 ° C. for 3 days followed by filtration. Nagasaru Ogase extract 858.3 g (evaporation residue 0.460 (w / v)%). The obtained Nagasaga Ogase extract was stored at ⁇ 5 ° C. for 12 days and then filtered to obtain 849.2 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before activated carbon treatment. About the Nagasaruogase extract before the activated carbon treatment obtained, the usnic acid content was measured by the same method as described above. The results are shown in Table 9.
  • Example 1-27 After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) 1.08 mg (manufactured by Enviro Chemicals) (10% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with 10 wt% of activated carbon.
  • activated carbon trade name: Shirasagi M, Japan
  • 1.08 mg manufactured by Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid content was measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 9.
  • Example 1-28 After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 5.40 mg (50 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon 50 wt%.
  • activated carbon trade name: Shirasagi M, Japan
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid content was measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 9.
  • Example 1-29 After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 10.80 mg (100% by weight based on solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with activated carbon 100 wt%.
  • activated carbon trade name: Shirasagi M, Japan
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid content was measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 9.
  • Example 1-30 After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 16.20 mg (150 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with 150 wt% of activated carbon.
  • activated carbon trade name: Shirasagi M, Japan
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid content was measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 9.
  • Example 1-31 After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd. (21.60 mg) (200% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon of 200 wt%.
  • activated carbon trade name: Shirasagi M, Japan
  • Enviro Chemicals Co., Ltd. 21.60 mg (200% by weight with respect to the solid content in the extract
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid content was measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 9.
  • Nagasaga Ogase extract is obtained by performing low-temperature storage treatment and activated carbon treatment on Nagasaga Ogase extract obtained using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent. Can further reduce the usnic acid content.
  • Example 1-32 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed and extracted at 20 ° C. for 8 days followed by filtration. Nagasaru Ogase extract 855.8 g (evaporation residue 0.270 (w / v)%). The obtained Nagasaga Ogase extract was stored at ⁇ 5 ° C. for 13 days and then filtered to obtain 839.5 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before hexane washing treatment. . About the Nagasagaogase extract before the obtained hexane washing process, the usnic acid content was measured by the method similar to the above. The results are shown in Table 10.
  • Example 1-33 Add 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution to 50 mL of Nagasaru Ogase extract obtained in Example 1-32 before washing with hexane, separate into two layers using a separatory funnel, and recover the lower layer Then, the usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 10.
  • Example 1-34 To 50 mL of Nagasaga Ogase extract obtained in Example 1-33, 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution were added again. After separating into two layers using a separatory funnel, the lower layer was recovered (hexane The usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 10.
  • Example 1-35 To 50 mL of Nagasaga Ogase extract obtained in Example 1-34, 50 mL of hexane and 5 mL of a 99.5% by volume ethanol aqueous solution were added again. After separating into two layers using a separatory funnel, the lower layer was recovered (hexane The product was washed three times), and the usnic acid content was measured by HPLC in the same manner as described above (evaporation residue 0.286 (w / v)%). The results are shown in Table 10.
  • Nagasagaogase extract is obtained by performing low-temperature storage treatment and hexane washing treatment on Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent.
  • the usnic acid content contained in the product can be further reduced.
  • Examples 1-36 to 1-43 10 mL of the Nagasaga Ogase extract after the hydrolyzing and low-temperature storage treatment obtained in Example 1-17 was stored at 60 ° C., and the Nagasaga Ogase extract after each storage time shown in Table 11 was the same as described above.
  • the usnic acid content was measured by HPLC according to the method. The results are shown in Table 11.
  • Examples 1-44 to 1-47 The same method as described above for the Nagasaga Ogase extract obtained by stirring 150 mL of Nagasaro Ogase extract obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment at 70 ° C. for the time shown in Table 11. Was used to measure the content of usnic acid by HPLC. The results are shown in Table 12.
  • Nagasagaogase extract obtained by using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and heat treatment.
  • the usnic acid content contained in the Nagasaru Ogase extract can be further reduced.
  • Example 1-51 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaga Ogase extract 175.62 g (evaporation residue 0 .290 (w / v)%).
  • Example 1-52 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and 174.25 g of Nagasal Ogase extract (evaporation residue 0 .340 (w / v)%).
  • Example 1-53 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 177.57 g (evaporation residue 0 .410 (w / v)%).
  • Example 1-54 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaga Ogase extract 178.19 g (evaporation residue 0 260 (w / v)%).
  • Example 1-55 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and Nagasal Ogase extract 179.82 g (evaporation residue 0 .300 (w / v)%).
  • Example 1-56 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 182.14 g (evaporation residue 0 .380 (w / v)%).
  • a Nagasaruogase extract having a sufficiently low concentration of usnic acid can be obtained.
  • the usnic acid content increased (Comparative Example 1-8).
  • a Salusose lichen extract containing a low amount of usnic acid can be produced by extracting the Ligosa lichen using an alcohol aqueous solution having an alcohol content of 80% by volume or less. Since usnic acid has hepatotoxicity, according to the first embodiment of the present invention, an extract of Nagasaga Ogase which is excellent in safety and can be suitably used for an external skin preparation such as an endothelin action inhibitor is produced. Can do.
  • Example 1-57 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract (177.6 g) 420 (w / v)%). 10 mL of the obtained extract was stored at ⁇ 5 ° C. for 5 days and then filtered to obtain 8.9 g of Nagasaga Ogase extract after evaporation at low temperature (evaporation residue 0.380 (w / v)%).
  • the Ussanoic acid content is further reduced by performing a low-temperature storage treatment on the Nagasagaogase extract obtained using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent. can do.
  • Example 1-58 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% 1,3-butanediol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration. 0.6 g (evaporation residue 0.420 (w / v)%) was obtained. Next, the mixture was diluted by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 1 mL of distilled water to perform starch, and a Nagasal Ogase extract after hydrolysis was obtained. Further, 9.5 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at ⁇ 5 ° C. for 5 days and then filtered, and 8.6 g of the Nagasaro Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.350 (w / v )%).
  • Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment.
  • the measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 16.
  • the theoretical value of the usnic acid content contained in the Nagasaru Ogase extract after the hydrotreatment is also shown in Table 16.
  • Example 1-59 Digestion was performed by mixing 10 mL of Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 1-58 and 2.5 mL of distilled water, and extraction was performed after Nagasal Ogase after hydrolysis. I got a thing. Further, 12 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at ⁇ 5 ° C. for 5 days and filtered, and then 10.5 g of the Nagasaga Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.300 (w / v)%) )
  • Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment.
  • the measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 17.
  • Table 17 also shows the theoretical value of the usnic acid content contained in the Nagasaru Ogase extract after the hydrotreatment.
  • Example 1-60 Distilled by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 1-58 and 5 mL of distilled water. Obtained. Furthermore, 14 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 5 days and filtered, and 12.6 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (evaporation residue 0.240 (w / v)%) )
  • Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment.
  • the measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 18.
  • the theoretical value of the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment is also shown in Table 18.
  • Example 1-61 Distillation was performed by mixing 10 mL of Nagasaru Ogase extract before hydrolysis and low-temperature storage prepared in the same manner as in Example 1-58 and 7.5 mL of distilled water, and extraction was performed after Nagasaga Ogase after hydrolysis. I got a thing. Further, 16 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 5 days and filtered, and 14.1 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.200 (w / v)%) )
  • Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment.
  • the measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 19.
  • Table 19 also shows the theoretical value of the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment.
  • Example 1-62 Distilled by mixing 10 mL of Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 1-58 and 10 mL of distilled water, and subjected to hydrolysis, Obtained. Further, 18 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 5 days and filtered, and then 15.9 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.170 (w / v )%).
  • Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment.
  • the measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 20.
  • the theoretical value of the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment is also shown in Table 20.
  • the Nagasaruogase extract obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to a hydrolysis treatment and a low-temperature storage treatment in this order.
  • the usnic acid content contained in the Nagasaru Ogase extract can be further reduced.
  • Example 1-63 Yokowa Sarugase (obtained from Hichem Corp.) and 10 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration.
  • Example 1-64 Yokowa Sarugase (obtained from Hichem) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 156.79 g (evaporation residue 0.402 (w / v)) %).
  • Example 1-65 Yokowa Sarugase (obtained from Hichem) 10 g and 50 vol% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 162.18 g (Evaporation residue 0.348 (w / v)) %).
  • Example 1-66 Yokowa Sarugase (obtained from Hichem Corp.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days followed by filtration. Yokowa Sarugase extract 166.61 g (evaporation residue 0.250) (W / v)%).
  • Example 1-67 Yokowa Sarugase (obtained from Hichem) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days, followed by filtration.
  • a Salusose lichen extract containing a low amount of usnic acid can be produced by extracting the Ligosa lichen using an aqueous alcohol solution having an alcohol content of 80% by volume or less. Since usnic acid has hepatotoxicity, according to the first embodiment of the present invention, it is possible to produce a Sarugose lichen that is excellent in safety and can be suitably used for an external skin preparation such as an endothelin action inhibitor. it can.
  • Example 1-68 88.57 g of the extract obtained from Example 1-64 (Yokosawa Ogase extract before hydrolyzing and low-temperature storage treatment) was stored at ⁇ 5 ° C. for 3 days and then filtered to obtain Yokosawa Ogase extract after low-temperature storage treatment. 81.68 g (evaporation residue 0.390 (w / v)%) was obtained. 80 mL of the obtained Yokosawa-ogase extract after low-temperature storage treatment and 20 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Yokosawa-ogase extract after the hydrotreatment. Further, the Yokosawa ogase extract after the hydrotreatment was stored at -5 ° C. for 4 days and then filtered, and 86.12 g (Yield residue 0.328 (w / v)%) of the Yokosawa ogase extract after the hydration / low temperature preservation treatment Got.
  • Example 1-69 40 mL of the Yokosawa Ogase extract obtained in Example 1-64 (Yokosawa Ogase extract before hydrolyzing and low-temperature storage treatment) and 10 mL of distilled water were mixed to dilute the ethanol volume to 40%, followed by hydrolysis. The later Yokosawa Ogase extract was obtained. Further, the Yokosawa ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 3 days and then filtered, and 41.03 g (Yield residue 0.286 (w / v)%) of the Yokosawa ogase extract after the hydrolyzing and low-temperature preservation treatment Got.
  • the usnic acid content was measured by the same method as described above. The results are shown in Table 23. In addition, Table 23 also shows the theoretical value of the usnic acid content contained in the Yokosawa ogase extract after the hydrotreatment.
  • the usnic acid content was measured by the same method as described above. The results are shown in Table 24.
  • the theoretical value of the usnic acid content contained in the Yokosawa ogase extract after the hydrotreatment is also shown in Table 24.
  • the Yokosawa ogase extract obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to a hydration treatment and a low-temperature storage treatment to thereby obtain a Yokosawa ogase.
  • the usnic acid content contained in the extract can be further reduced.
  • Example 1-71 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of 5 volume% ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 96.98 g of filtrate (evaporation residue 0.080 (w / v )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue. 32.48 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration, and Nagasaga Ogase extract 178.44 g (evaporation residue 0.170 (w / v)%) )
  • Example 1-72 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of a 1% by volume ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 959.01 g of filtrate (evaporation residue 0.076 (w / v )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue. 35.47 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days followed by filtration. The Nagasaga Ogase extract 180.03 g (evaporation residue 0.152 (w / v)%) )
  • Example 1-73 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of distilled water were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 950.01 g of filtrate (evaporation residue 0.080 (w / v)%) Separated. Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue. 32.75 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration. The Nagasaga Ogase extract 179.03 g (evaporation residue 0.144 (w / v)% )
  • the Usunic acid content is particularly improved by treating the Sarcophagus lichen with an aqueous solvent before extraction with an aqueous alcohol solution having an alcohol content of 80% by volume or less.
  • a low Sargogassian lichen extract can be obtained.
  • Example 1-74 80 mL of the extract obtained in Example 1-71 (Nagasaruogase extract before hydrolyzing and low-temperature storage treatment) and 20 mL of distilled water were mixed to dilute the ethanol volume to about 40-50%. The Nagasagaogase extract after the hydration treatment was obtained. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 4 days and filtered, and 84.33 g of the Nagasao Ogase extract after the hydrous and low temperature preservation treatment (evaporation residue 0.140 (w / v)%) Got.
  • Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment. The results are shown in Table 26. In addition, the theoretical value of the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment is also shown in Table 26.
  • Nagasaga Ogase extract is obtained by subjecting Nagasaga Ogase extract obtained by using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent to hydrotreating and low-temperature storage treatment.
  • the usnic acid content contained in can be selectively reduced.
  • Example 2-1 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 30 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 199.90 g (evaporation residue 0.146 (w / v)%).
  • Example 2-2 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 40 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasal Ogase extract 200.70 g (evaporation residue 0.190 (w / v)%).
  • Example 2-3 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 5 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.67 g (evaporation residue 0.178 (w / v)%).
  • Example 2-4 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 15 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 202.57 g (evaporation residue 0.192 (w / v)%).
  • Example 2-5 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.69 g (evaporation residue 0.298 (w / v)%).
  • Example 2-6 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 30 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 200.54 g (evaporation residue 0.258 (w / v)%).
  • Example 2-7 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.49 g (evaporation residue 0.316 (w / v)%).
  • Example 2-8 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 60 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 198.69 g (evaporation residue 0.242 (w / v)%).
  • Example 2-9 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 70 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, 195.1 g of Nagasal Ogase extract (evaporation residue 0.232 (w / v)%).
  • Example 2-10 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 80 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.6 g (evaporation residue 0.216 (w / v)%).
  • Difractic acid content and usnic acid content in each Nagasagaogase extract obtained in Examples 2-1 to 2-10 and Comparative Examples 2-1 to 2-11 The amount was analyzed using HPLC under the following conditions.
  • the apparatus and measurement conditions used for the measurement are as follows.
  • Measuring instruments HPLC: Agilent Technologies Open LAB CDS LC02-1260, Autosampler G1329B 1260 ALS, Column oven G1316A 1260 TCC, Detector G1315D 1260 DAD VL, Pump G1312B 1260 Bin pump, G1379B 1260 ⁇ -Degasser ⁇ HPLC analysis conditions> Flow rate: 0.75 mL / min Column used: Inertsil ODS-3 5 ⁇ m (GL Science), 3.0 ID ⁇ 150 mm Detection wavelength: 254 nm Column temperature: 40 ° C Mobile phase: A: 0.1% trifluoroacetic acid-containing aqueous solution, B: 0.1% trifluoroacetic acid-containing methanol 0 to 10 minutes: 70% B 10-15 minutes: 70% B ⁇ 100% B 15-20 minutes: 100% B The results are shown in Table 27.
  • the difractic acid content was sufficiently high and the usnic acid content was reduced, and the difractic acid high content and the usnic acid low content were low.
  • Nagasarogase extract can be obtained.
  • the usnic acid content decreased, but the difractic acid content also significantly decreased (Comparative Examples 2-1 to 2-4).
  • the content of difractate increased, but the content of usnic acid also increased (Comparative Examples 2-5 to 2-11).
  • an extract of Salgogase lichen having a high content of difractate and a low content of usnic acid is produced by extracting the Licorice lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume.
  • Usnic acid has hepatotoxicity, while difractic acid has an excellent endothelin inhibitory action. Therefore, according to the second embodiment of the present invention, an endothelin action inhibitor having an endothelin action inhibitory effect and excellent in safety can be produced.
  • Example 2-11 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasaga Ogase extract 858.5 g (evaporation residue 0.374 (w / v)%). The obtained extract was stored at ⁇ 5 ° C. for 12 days and then filtered to obtain 833.4 g of Nagasaroogase extract (low evaporation residue 0.360 (w / v)%) after the low-temperature storage treatment.
  • Nagasagaogase extract is obtained by subjecting Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent to a low-temperature storage treatment.
  • the usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the product.
  • Example 2-12 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasal Ogase extract 852.8 g (evaporation residue 0.268 (w / w v)%). The obtained extract was stored at ⁇ 5 ° C. for 13 days and filtered to obtain 837.2 g (evaporation residue 0.256 (w / v)%) of the Nagasaga Ogase extract before treatment. Next, 10 mL of Nagasaga Ogase extract pretreatment product and 2.5 mL of distilled water were mixed and diluted so that the volume of ethanol was 40%. .
  • the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 4 days and then filtered, and the Nagasal Ogase extract after the hydrous / low temperature preservation treatment was 11.1 g (evaporation residue 0.130 (w / v)%).
  • Nagasagaogase extract obtained using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to low-temperature storage treatment, hydrolysis treatment and low-temperature storage treatment.
  • the usnic acid content contained in the Nagasaga Ogase extract can be selectively reduced.
  • Example 2-13 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 1 L of 50% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 7 days, filtered, and 838.5 g (Evaporation residue) Min 0.218 (w / v)%).
  • 800 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 200 mL of distilled water are mixed and diluted so that the volume of ethanol becomes 40%, and then the Nagasaga Ogase extract after hydrolysis is obtained. It was. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 7 days and filtered, and 916.2 g of the Nagasal Ogase extract after the hydrothermal treatment at low temperature (Evaporation residue 0.130 (w / v)%) Got.
  • the difractic acid content and the usnic acid content were measured by the same method as described above. The results are shown in Table 6. In addition, the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment are also shown in Table 30.
  • the Nagasaruogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a hydration treatment and a low-temperature storage treatment.
  • the usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the Nagasaru Ogase extract.
  • Example 2-14 5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. Then, add 0.65 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (10% by weight based on the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 10 wt% -treated product was obtained.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.08 (w / v)%)). The results are shown in Table 31.
  • Example 2-15 5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. After dilution, add 3.25 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (50% by weight based on the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 50 wt% treated product was obtained.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
  • Example 2-16 5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. After dilution, add 6.50 mg of activated carbon (trade name: Shirahige M, manufactured by Nippon Enviro Chemicals) (100% by weight with respect to the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 100 wt% treated product was obtained.
  • activated carbon trade name: Shirahige M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
  • Example 2-17 5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. Then, 9.75 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (150% by weight with respect to the solid content in the extract) is added, and the mixture is stirred at room temperature for 45 minutes and then filtered. Thus, a product after treatment with 150 wt% of activated carbon was obtained.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
  • Example 2-18 5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. 13.0 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (200% by weight with respect to the solid content in the extract) is added, stirred for 45 minutes at room temperature, and then filtered. Thus, a product after treatment with 200 wt% of activated carbon was obtained.
  • activated carbon trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals
  • the obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
  • the Nagasagaogase extract obtained using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and activated carbon treatment.
  • the usnic acid content can be selectively reduced while maintaining the content of difractacic acid contained in the Nagasaroogase extract.
  • Example 2-19 50 mL of hexane and 5 mL of 99.5 vol% ethanol aqueous solution were added to 50 mL of Nagasaroogase extract (extract before hexane washing treatment) obtained in Example 2-13 after hydrolyzing and low-temperature storage treatment, and a separatory funnel was used. After separating into two layers, the lower layer was recovered (hexane washed once product), and the contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 32.
  • Example 2-20 To 50 mL of Nagasaga Ogase extract obtained in Example 2-19, 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution were added again, and after separating into two layers using a separatory funnel, the lower layer was recovered (hexane The contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 32.
  • Example 2-21 Add 45 mL of hexane again to 45 mL of Nagasaga Ogase extract obtained in Example 2-20, separate into two layers using a separatory funnel, and recover the lower layer (hexane washed three times), as above
  • the contents of difractic acid and usnic acid were measured by HPLC according to the above method. The results are shown in Table 32.
  • Nagasagaogase extract obtained by using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to hydration treatment, low-temperature storage treatment, and hexane washing treatment.
  • hydration treatment low-temperature storage treatment
  • hexane washing treatment it is possible to selectively reduce the usnic acid content while maintaining the content of difractic acid contained in the Nagasaroogase extract.
  • Example 2-22 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% aqueous ethanol solution 1 L were mixed and extracted at 20 ° C. for 3 days followed by filtration. Nagasaru Ogase extract 858.3 g (evaporation residue 0.460 (w / v)%). The obtained Nagasaga Ogase extract was stored at ⁇ 5 ° C. for 12 days and then filtered to obtain 849.2 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before activated carbon treatment. About the obtained Nagasaruogase extract before the activated carbon treatment, the difractic acid content and the usnic acid content were measured by the same method as described above. The results are shown in Table 33.
  • Example 2-23 27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) 1.08 mg (manufactured by Enviro Chemicals) (10% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with 10 wt% of activated carbon.
  • activated carbon trade name: Shirasagi M, Japan
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 33.
  • Example 2-24 27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 5.40 mg (50 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon 50 wt%.
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 33.
  • Example 2-25 27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 10.80 mg (100% by weight based on solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with activated carbon 100 wt%.
  • activated carbon trade name: Shirasagi M, Japan
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 33.
  • Example 2-26 27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 16.20 mg (150 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with 150 wt% of activated carbon.
  • activated carbon trade name: Shirasagi M, Japan
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 33.
  • Example 2-27 27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd. (21.60 mg) (200% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon of 200 wt%.
  • activated carbon trade name: Shirasagi M, Japan
  • the obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment.
  • the acid and usnic acid contents were measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 33.
  • Example 2-28 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed and extracted at 20 ° C. for 8 days followed by filtration. Nagasaru Ogase extract 855.8 g (evaporation residue 0.270 (w / v)%). The obtained Nagasaga Ogase extract was stored at ⁇ 5 ° C. for 13 days and then filtered to obtain 839.5 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before hexane washing treatment. . About the Nagasaruogase extract before the obtained hexane washing process, the difractic acid content and the usnic acid content were measured by the method similar to the above. The results are shown in Table 34.
  • Example 2-29 Add 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution to 50 mL of Nagasaga Ogase extract obtained in Example 2-28 before washing with hexane, separate into two layers using a separatory funnel, and recover the lower layer Then, the contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 34.
  • Example 2-30 To 50 mL of Nagasaga Ogase extract obtained in Example 2-29, 50 mL of hexane and 5 mL of a 99.5% by volume ethanol aqueous solution were added again. After separating into two layers using a separatory funnel, the lower layer was recovered (hexane The contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 34.
  • Example 2-31 To 50 mL of Nagasaga Ogase extract obtained in Example 2-30, 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution were added again, and after separating into two layers using a separatory funnel, the lower layer was recovered (hexane The contents of difractic acid and usnic acid were measured by HPLC by the same method as described above (evaporation residue 0.286 (w / v)%). The results are shown in Table 34.
  • Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a low-temperature storage treatment and a hexane washing treatment.
  • the usnic acid content can be selectively reduced without significantly losing the content of difractic acid contained in the Nagasaru Ogase extract.
  • Examples 2-32 to 2-39 10 mL of the Nagasaga Ogase extract after hydrolyzed and cryopreserved treatment obtained in Example 2-13 was stored at 60 ° C., and the Nagasaga Ogase extract after each storage time shown in Table 35 was the same as described above.
  • the contents of difractic acid and usnic acid were measured by HPLC according to the method. The results are shown in Table 35.
  • Examples 2-40 to 2-43 150 mL of the Nagasaga Ogase extract obtained after the hydrolyzing and low-temperature storage treatment obtained in Example 2-13 was stirred at 70 ° C. for the time shown in Table 12, and the obtained Nagasaga Ogase extract was subjected to the same method as described above. Was used to measure the contents of difractic acid and usnic acid by HPLC. The results are shown in Table 36.
  • the Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and heat treatment.
  • the usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the Nagasaruogase extract.
  • Example 2-47 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaga Ogase extract 175.62 g (evaporation residue 0 .290 (w / v)%).
  • Example 2-48 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and 174.25 g of Nagasal Ogase extract (evaporation residue 0 .340 (w / v)%).
  • Example 2-49 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 177.57 g (evaporation residue 0 .410 (w / v)%).
  • Example 2-51 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and Nagasal Ogase extract 179.82 g (evaporation residue 0 .300 (w / v)%).
  • Example 2-52 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 182.14 g (evaporation residue 0 .380 (w / v)%).
  • the difractic acid content was sufficiently high and the usnic acid content was reduced, and the difractic acid high content and the low usnic acid content were low.
  • Nagasarogase extract can be obtained.
  • the concentration of difractate increased, but the concentration of usnic acid also increased (Comparative Example 2-12).
  • the extract of Salogase lichen having a high content of difractic acid and a low content of usnic acid is produced by extracting the Licorice lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume.
  • Example 2-53 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract (177.6 g) 420 (w / v)%). 10 mL of the obtained extract was stored at ⁇ 5 ° C. for 5 days and then filtered to obtain 8.9 g of Nagasaga Ogase extract after evaporation at low temperature (evaporation residue 0.380 (w / v)%).
  • Nagasaga Ogase extract is obtained by subjecting Nagasaga Ogase extract obtained by using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent to a low-temperature storage treatment.
  • the usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the product.
  • Example 2-54 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% 1,3-butanediol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration. 0.6 g (evaporation residue 0.420 (w / v)%) was obtained. Next, the mixture was diluted by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 1 mL of distilled water to perform starch, and a Nagasal Ogase extract after hydrolysis was obtained. Further, 9.5 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at ⁇ 5 ° C. for 5 days and then filtered, and 8.6 g of the Nagasaro Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.350 (w / v )%).
  • Example 2-55 Digestion was performed by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 2.5 mL of distilled water prepared in the same manner as in Example 2-54, and Nagasaga Ogase extraction after hydrolysis was performed. I got a thing. Further, 12 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at ⁇ 5 ° C. for 5 days and filtered, and then 10.5 g of the Nagasaga Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.300 (w / v)%) )
  • Example 2-56 Distilled by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 2-54 and 5 mL of distilled water, and then subjected to hydrolysis. Obtained. Furthermore, 14 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 5 days and filtered, and 12.6 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (evaporation residue 0.240 (w / v)%) )
  • Example 2-57 Digestion was performed by mixing 10 mL of Nagasaga Ogase extract before hydrolysis / low-temperature storage treatment prepared in the same manner as in Example 2-54 and 7.5 mL of distilled water, and extraction was performed after Nagasaga Ogase after hydrolysis. I got a thing. Further, 16 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 5 days and filtered, and 14.1 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.200 (w / v)%) )
  • Example 2-58 Distilled by mixing 10 mL of Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 2-54 and 10 mL of distilled water, and then subjected to hydrolysis. Obtained. Further, 18 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 5 days and filtered, and then 15.9 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.170 (w / v )%).
  • Example 2-59 Yokowa Sarugase (obtained from Hichem) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 156.79 g (evaporation residue 0.402 (w / v)) %).
  • Example 2-60 Yokowa Sarugase (obtained from Hichem) 10 g and 50 vol% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 162.18 g (Evaporation residue 0.348 (w / v)) %).
  • Example 2-61 Yokowa Sarugase (obtained from Hichem Corp.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days followed by filtration.
  • Yokowa Sarugase extract 166.61 g evaporation residue 0.250) (W / v)%).
  • Example 2-62 Yokowa Sarugase (obtained from Hichem) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days, followed by filtration.
  • the difractic acid content was sufficiently high and the usnic acid content was reduced, and the difractic acid content was high and the usnic acid content was low.
  • Yokowa Sarugase extract can be obtained.
  • Yokosawa ogase was extracted using an extraction solvent having an alcohol content that was too low, the usnic acid content decreased, but the difractic acid content also significantly decreased (Comparative Example 2-12).
  • Yokowa sucrose was extracted using an extraction solvent having an alcohol content that was too high, the difractic acid content increased, but the usnic acid content also increased (Comparative Examples 2-13 and 2-14).
  • an extract of Salgogase lichen having a high content of difractate and a low content of usnic acid is produced by extracting the Licorice lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume.
  • Usnic acid has hepatotoxicity, while difractic acid has an excellent endothelin inhibitory action. Therefore, according to the 2nd embodiment of this invention, the endothelin action inhibitor which has an endothelin action inhibitory effect and was excellent in safety
  • security can be manufactured.
  • Example 2-63 88.57 g of the extract obtained in Example 2-59 (Yokosawa ogousse extract before hydrolyzing and low-temperature storage treatment) was stored at ⁇ 5 ° C. for 3 days and filtered to obtain a Yokosawa ogousse extract after low-temperature storage treatment. 81.68 g (evaporation residue 0.390 (w / v)%) was obtained. 80 mL of the obtained Yokosawa-ogase extract after low-temperature storage treatment and 20 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Yokosawa-ogase extract after the hydrotreatment. Further, the Yokosawa ogase extract after the hydrotreatment was stored at -5 ° C. for 4 days and then filtered, and 86.12 g (Yield residue 0.328 (w / v)%) of the Yokosawa ogase extract after the hydration / low temperature preservation treatment Got.
  • Example 2-64 40 mL of the Yokosawa Ogase extract obtained in Example 2-59 (Yokosawa Ogase extract before hydrolyzing and low-temperature storage treatment) and 10 mL of distilled water were mixed to dilute the ethanol volume to 40%, followed by hydrolysis. The later Yokosawa Ogase extract was obtained. Further, the Yokosawa ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 3 days and then filtered, and 41.03 g (Yield residue 0.286 (w / v)%) of the Yokosawa ogase extract after the hydrolyzing and low-temperature preservation treatment Got.
  • the Yokosawa ogase extract obtained by using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a hydrolysis treatment and a low-temperature storage treatment.
  • the usnic acid content can be selectively reduced while maintaining the content of difractacic acid contained in the Yokosawa ogase extract.
  • Example 2-66 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of 5 volume% ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 96.98 g of filtrate (evaporation residue 0.080 (w / v )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue. 32.48 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration, and Nagasaga Ogase extract 178.44 g (evaporation residue 0.170 (w / v)%) )
  • Example 2-67 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of a 1% by volume ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration. )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue. 35.47 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days followed by filtration. The Nagasaga Ogase extract 180.03 g (evaporation residue 0.152 (w / v)%) )
  • Example 2-68 Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of distilled water were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 950.01 g of filtrate (evaporation residue 0.080 (w / v)%) Separated. Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue. 32.75 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration. The Nagasaga Ogase extract 179.03 g (evaporation residue 0.144 (w / v)% )
  • Example 2-69 80 mL of the extract obtained in Example 2-66 (Nagasaruogase extract before hydrolyzing and low-temperature storage treatment) and 20 mL of distilled water were mixed to dilute the ethanol volume to about 40 to 50%.
  • the Nagasagaogase extract after the hydration treatment was obtained. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 4 days and filtered, and 84.33 g of the Nagasao Ogase extract after the hydrous and low temperature preservation treatment (evaporation residue 0.140 (w / v)%) Got.
  • the Nagasaruogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a hydration treatment and a low-temperature storage treatment.
  • the usnic acid content contained in the Nagasaga Ogase extract can be selectively reduced.
  • Example 3-1 The Nagasal Ogase extract obtained by Comparative Example 3-7 before the hydrothermal treatment at low temperature and 8 mL of distilled water and 11 mL of distilled water were mixed to dilute the ethanol volume to 40%. Got. Furthermore, the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 12 days and then filtered, and the Nagasal Ogase extract after the hydrous and low temperature preservation treatment 13.5 g (evaporation residue 0.084 (w / v)%) Got.
  • Table 52 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment. Furthermore, Table 52 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaru Ogase before the hydrothermal treatment at low temperature.
  • Example 3-2 The Nagasal Ogase extract obtained in Comparative Example 3-8 before the hydrolysis / low-temperature storage treatment was mixed with 8 mL of distilled water and 11 mL of distilled water to dilute the ethanol volume to 40%. Got. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 12 days and then filtered, and the Nagasal Ogase extract after the hydrous and low temperature preservation treatment 13.4 g (evaporation residue 0.114 (w / v)%) Got.
  • Table 53 shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment. Furthermore, Table 53 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
  • Example 3-3 The Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-9 was stored at ⁇ 5 ° C. for 37 days and then filtered to obtain 140.1 g of Nagasaro Ogase extract after low-temperature storage treatment (evaporation residue). 0.136 (w / v)%) was obtained.
  • the obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
  • the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 6 days and then filtered, and the Nagasaga Ogase extract after the hydrous / low temperature preservation treatment 206.0 g (evaporation residue 0.06 (w / v)%) Got.
  • Example 3-4 The Nagasaga Ogase extract obtained in Comparative Example 3-10, which had not been hydrolyzed and stored at a low temperature, was stored at ⁇ 5 ° C. for 37 days and then filtered to obtain 137.2 g of the Nagasal Ogase extract after the low temperature storage (the evaporation residue). Min 0.166 (w / v)%).
  • the obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 6 days and then filtered, and then 204.3 g of the Nagasaga Ogase extract after the hydrous and low temperature preservation treatment (evaporation residue 0.05 (w / v)%) Got.
  • Nagasaga Ogase extract before hydrolyzed and cryopreserved Nagasal ogase extract after cryopreserved
  • Nagasal ogused extract after hydrolyzed and Nagasal ogused extract after hydrolyzed and cryopreserved was used to measure the difractic acid content and the usnic acid content.
  • the results are shown in Table 55.
  • the theoretical value of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment is also shown in Table 55.
  • Table 55 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
  • Example 3-5 The Nagasaga Ogase extract obtained in Comparative Example 3-11 before the hydrothermal and low temperature preservation treatment was stored at ⁇ 5 ° C. for 33 days and filtered to obtain 139.7 g of the Nagasal Ogase extract after the low temperature preservation treatment (evaporation residue). Min 0.196 (w / v)%).
  • the obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
  • the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 3 days and then filtered, and the Nagasaga Ogase extract after the hydrous / low temperature preservation treatment 205.7 g (evaporation residue 0.06 (w / v)%) Got.
  • Nagasaroogase extract before hydrolyzing and low-temperature preservation treatment Nagasaroogase extract after cryopreservation treatment, Nagasaroogase extract after hydrolyzing treatment, and Nagasaroogase extract after hydrolyzing and cryopreservation treatment was used to measure the difractic acid content and the usnic acid content.
  • Table 56 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment.
  • Table 56 also shows the residual rate of usnic acid relative to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
  • Example 3-6 133.61 g of Nagasaga Ogase extract after low-temperature storage treatment was filtered by storing 139.62 g of Nagasaro-ogase extract before hydrolysis / low-temperature storage treatment obtained in Comparative Example 3-12 at -5 ° C. for 3 days. (Evaporation residue 0.248 (w / v)%) was obtained.
  • the obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
  • the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 3 days and then filtered, and 201.58 g of the Nagasal Ogase extract after the hydrothermal treatment at low temperature (evaporation residue 0.09 (w / v)%) Got.
  • Example 3-7 141.50 g of the Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-13 was stored at ⁇ 5 ° C. for 2 days and filtered to obtain 132.31 g of Nagasaga Ogase extract after low-temperature storage treatment. (Evaporation residue 0.358 (w / v)%) was obtained.
  • the obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
  • the Nagasaga Ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 3 days and filtered, and 197.0 g of the Nagasal Ogase extract after the hydrous and low-temperature storage treatment (evaporation residue 0.104 (w / v)%) Got.
  • Nagasaga Ogase extract before hydrolyzed and cryopreserved Nagasal ogase extract after cryopreserved
  • Nagasal ogused extract after hydrolyzed and Nagasal ogused extract after hydrolyzed and cryopreserved was used to measure the difractic acid content and the usnic acid content.
  • Table 58 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment.
  • Table 58 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
  • the extract of lichenaceae lichen using only an alcohol aqueous solution with an alcohol content of more than 70% by volume as the extraction solvent has a very high usnic acid content in the extract. high.
  • the usnic acid content can be selectively reduced by performing a selective reduction process of usnic acid.
  • Example 3-8 Extract 81.05 g of the extract obtained in Comparative Example 3-17 (Yokosawa-o-gase extract before hydrolyzing and low-temperature storage treatment) was stored at ⁇ 5 ° C. for 3 days, and then filtered to extract Yokosawa-o-gase after low-temperature storage treatment. As a result, 75.02 g (0.272 (w / v)% of evaporation residue) of the product was obtained. The obtained Yokosawa ogase extract after low-temperature storage treatment and 68.75 mL of distilled water were mixed and diluted so that the volume of ethanol was 40%, and the Yokosawa ogase extract after the hydration treatment was obtained.
  • the Yokosawa ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 4 days and then filtered, and 98.97 g of the Yokosawa ogase extract after the hydrothermal treatment at low temperature (evaporation residue 0.08 (w / v)%) Got.
  • Example 3-9 50 ml of the extract of Yokosawa-o-gase before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-19 and 68.75 mL of distilled water were mixed to dilute the ethanol volume to 40%. An extract was obtained. Further, the Yokosawa ogase extract after the hydrolysis treatment was stored at ⁇ 5 ° C. for 2 days and filtered, and 101.15 g of the Yokosawa ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.062 (w / v)%) Got.
  • Example 3-10 50 mL of the extract of Yokosawa-o-gase before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-20 and 68.75 mL of distilled water were mixed to dilute the ethanol volume to 40%. An extract was obtained. Further, the Yokosawa ogase extract after the hydrotreatment was stored at ⁇ 5 ° C. for 2 days and then filtered, and 100.15 g of the Yokosawa ogase extract after the hydrothermal treatment at low temperature (evaporation residue 0.102 (w / v)%) Got.
  • the extract of lichenaceae lichen using only an alcohol aqueous solution with an alcohol content of more than 70% by volume as the extraction solvent has a very high usnic acid content in the extract. high.
  • the usnic acid content can be selectively reduced by performing a selective reduction process of usnic acid.
  • Whitening agents in the form of lotions, emulsions, cosmetic liquids and creams having the composition shown below, each of which contains the extract obtained in the above example as an active ingredient, can be prepared by conventional methods.

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Abstract

A method for producing an extract from a lichen belonging to the genus Usnea which has a low usnic acid content, said method comprising extracting a lichen belonging to the genus Usnea with an aqueous alcohol solution having an alcohol content of 80 vol% or less.

Description

サルオガセ属地衣類抽出物の製造方法Method for producing Sargogase lichen extract
　本発明は、サルオガセ（Usnea）属地衣類抽出物の製造方法に関する。 The present invention relates to a method for producing a lichen extract of the genus Usnea .
　エンドセリンは、内皮細胞由来のペプチドホルモンで、受容体を通して種々の細胞や組織に作用する。例えば、血管平滑筋細胞等において細胞内カルシウム濃度上昇を引き起こすことが知られている（非特許文献１）。
　近年、エンドセリンが表皮メラノサイト（メラニン細胞）に対し細胞内カルシウム濃度上昇を促し、細胞内のシグナル伝達系を介して細胞増殖を促進するとともに、メラニン合成の律速酵素であるチロシナーゼの活性を増強することが報告されている（非特許文献２参照）。また、エンドセリンは表皮角化細胞（ケラチノサイト）が産生するメラノサイト活性化因子の１つであること（非特許文献３）、紫外線誘導性色素沈着や老人性色素斑形成の重要な要因であることが報告されている（非特許文献４、５）。
　このようなエンドセリンの生体作用から、エンドセリンの作用を抑制しうる物質は、メラニン生成や色素沈着等を抑制し、美白剤として有用である。 Endothelin is a peptide hormone derived from endothelial cells and acts on various cells and tissues through receptors. For example, it is known to cause an increase in intracellular calcium concentration in vascular smooth muscle cells and the like (Non-Patent Document 1).
In recent years, endothelin has promoted an increase in intracellular calcium concentration in epidermal melanocytes (melanocytes), promoted cell proliferation via intracellular signal transduction system, and enhanced the activity of tyrosinase, the rate-limiting enzyme of melanin synthesis. Has been reported (see Non-Patent Document 2). In addition, endothelin is one of the melanocyte activators produced by epidermal keratinocytes (keratinocytes) (Non-patent Document 3), and is an important factor in ultraviolet-induced pigmentation and senile pigment spot formation. It has been reported (Non-Patent Documents 4 and 5).
A substance capable of suppressing the action of endothelin due to the biological action of endothelin suppresses melanin production and pigmentation and is useful as a whitening agent.
　一方、ウメノキゴケ（Parmeliaceae）科サルオガセ属の地衣類の抽出物がメラニン生成阻害作用を有することが知られている（特許文献１～３、並びに非特許文献６参照）。なお、特許文献１～３には、メラニン生成阻害物質の製造方法として、地衣類を培養し、その培養物を凍結乾燥した後０℃～１５℃の温度で溶媒抽出することが記載されている。 On the other hand, it is known that lichen extracts of the genus Sarogase belonging to the family Parmeliaceae have an inhibitory action on melanin production (see Patent Documents 1 to 3 and Non-Patent Document 6). In Patent Documents 1 to 3, as a method for producing a melanin production inhibitor, lichen is cultured, and the culture is freeze-dried and then subjected to solvent extraction at a temperature of 0 ° C. to 15 ° C. .
特開平４－２８２３１９号公報JP-A-4-282319 特開平４－２８２３２０号公報JP-A-4-282320 特開平４－２８２３２１号公報JP-A-4-282321
　本発明は、サルオガセ属地衣類を、アルコール含有率が８０体積％以下のアルコール水溶液を用いて抽出する、ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法に関する。
　また、本発明は、前記製造方法により得られるサルオガセ属地衣類抽出物を有効成分とする、エンドセリン作用抑制剤に関する。
　また、本発明は、前記製造方法により得られるサルオガセ属地衣類抽出物を有効成分とする、美白剤に関する。
（以下、前記ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法、前記エンドセリン作用抑制剤、及び美白剤を併せて本発明の第１の実施態様という。） TECHNICAL FIELD The present invention relates to a method for producing an extract containing Salusose lichen belonging to a low content of usnic acid, wherein the lichenaceae lichen is extracted using an alcohol aqueous solution having an alcohol content of 80% by volume or less.
In addition, the present invention relates to an endothelin action inhibitor comprising as an active ingredient the extract of the genus Sarhogase lichen obtained by the above production method.
The present invention also relates to a whitening agent comprising, as an active ingredient, an extract from the genus Sarugagase obtained by the production method.
(Hereinafter, the method for producing the Ususic acid low-containing Salogase lichen extract, the endothelin action inhibitor, and the whitening agent are collectively referred to as the first embodiment of the present invention.)
　また、本発明は、サルオガセ属地衣類を、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出する、ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法に関する。
　また、本発明は、サルオガセ属地衣類を、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出する、エンドセリン作用抑制剤の製造方法に関する。
　さらに、本発明は、前記製造方法により得られるサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤を有効成分とする、美白剤に関する。
（以下、前記ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法、前記エンドセリン作用抑制剤の製造方法、及び美白剤を併せて本発明の第２の実施態様という。） In addition, the present invention relates to a method for producing an extract of Sarugagase lichen having a high content of difractic acid and a low content of usnic acid, wherein the lichenaceae lichen is extracted using an alcohol aqueous solution having an alcohol content of 30% by volume to 80% by volume. .
In addition, the present invention relates to a method for producing an endothelin action inhibitor, which extracts Saruogase lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume.
Furthermore, the present invention relates to a whitening agent comprising, as an active ingredient, a Sargoses genus lichen extract or endothelin action inhibitor obtained by the production method.
(Hereinafter, the manufacturing method of the extract containing high amount of difractic acid and low content of usnic acid, the method of manufacturing the endothelin action inhibitor, and the whitening agent are collectively referred to as the second embodiment of the present invention.)
　また、本発明は、サルオガセ属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う抽出物の製造方法であって、前記処理後のウスニン酸の残存率が５０％未満である、ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法に関する。
　また、本発明は、サルオガセ属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う、ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法に関する。
　また、本発明は、サルオガセ属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う、エンドセリン作用抑制剤の製造方法に関する。
　さらに、本発明は、前記製造方法により得られるサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤を有効成分とする、美白剤に関する。
（以下、前記ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法、前記ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法、前記エンドセリン作用抑制剤の製造方法、及び美白剤を併せて本発明の第３の実施態様という。） In addition, the present invention is a method of extracting Sarugagase lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment. It is the manufacturing method of the extract which performs the above process, Comprising: It is related with the manufacturing method of the Ususic acid low content Salogase genus lichen extract which the residual rate of the usnic acid after the said processing is less than 50%.
In addition, the present invention is a method of extracting Sarugagase lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment. The present invention relates to a method for producing an extract of Lichenaceae lichen with a high content of difractic acid and a low content of usnic acid.
In addition, the present invention is a method of extracting Sarugagase lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment. It is related with the manufacturing method of the endothelin action inhibitor which performs the above process.
Furthermore, the present invention relates to a whitening agent comprising, as an active ingredient, a Sargoses genus lichen extract or endothelin action inhibitor obtained by the production method.
(Hereinafter, the method for producing the low extract of Usugic acid Salogase genus lichen, the method for producing the extract of Salgogase genus liquor containing the high content of difractate and low content of Usnic acid, the method of producing the endothelin action inhibitor, and the whitening agent) This is referred to as a third embodiment of the present invention.)
　ここで、特に断わらない限り、本発明とは前記第１の実施態様、第２の実施態様、及びの第３の実施態様のいずれも包含する意味である。 Here, unless otherwise specified, the present invention is meant to include all of the first embodiment, the second embodiment, and the third embodiment.
発明の詳細な説明Detailed Description of the Invention
　本発明は、美白剤などの皮膚外用剤に好適に用いることができるサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤の製造方法の提供を課題とする。 An object of the present invention is to provide a method for producing a Sargogase lichen extract or an endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent.
　本発明者らは、サルオガセ属地衣類抽出物に含まれる成分の検討、並びにサルオガセ属地衣類抽出物からの美白成分の探求と、当該成分を高濃度に含有しうる抽出方法について鋭意検討を行った。その結果、サルオガセ属地衣類の抽出物に含まれ、下記式で表されるジフラクタ酸が、エンドセリンの作用によって引き起こされるメラノサイト内のカルシウム（イオン）濃度上昇の抑制効果を有することを見出した。 The inventors of the present invention have conducted intensive investigations on the components contained in the extract of Lichenaceae lichens, the search for whitening components from Lichenaceae lichen extracts, and the extraction methods that can contain these components in high concentrations. As a result, it has been found that difractic acid, which is contained in the extract of the genus Sarhogase and represented by the following formula, has an inhibitory effect on the increase in calcium (ion) concentration in melanocytes caused by the action of endothelin.
Figure JPOXMLDOC01-appb-I000001
Figure JPOXMLDOC01-appb-I000001
　一方で、本発明者らは、サルオガセ属地衣類抽出物に含まれる成分の検討を行った結果、抽出方法によっては、サルオガセ属地衣類抽出物に下記式で表されるウスニン酸が高濃度で含まれることも見出した。 On the other hand, as a result of studying the components contained in the extract of the genus Sarhogase, the present inventors, depending on the extraction method, contain a high concentration of usnic acid represented by the following formula in the extract of the genus Sarogase genus. I also found out.
Figure JPOXMLDOC01-appb-I000002
Figure JPOXMLDOC01-appb-I000002
　前記ウスニン酸は、肝毒性物質として知られている（例えば、Frankos　VH（January　2005），"NTP　nomination　for　usnic　acid　and　Usnea　barbata　herb"，National　Toxicology　Program，http://ntp.niehs.nih.gov/ntp/htdocs/Chem_Background/ExSumPdf/UsnicAcid.pdf参照）。したがって、ジフラクタ酸の含有量が高くても、ウスニン酸の含有量も高いサルオガセ属地衣類抽出物は、安全性の観点から美白剤などの皮膚外用剤に用いることは問題である。 Usnic acid is known as a hepatotoxic substance (for example, Frankos VH (January 2005), "NTP nomination for usnic acid and Usnea barbata herb", National Toxicology Program, http://ntp.niehs.nih.gov /ntp/htdocs/Chem_Background/ExSumPdf/UsnicAcid.pdf). Therefore, it is a problem to use a Salgogase lichen extract having a high content of difractic acid and a high content of usnic acid in a skin external preparation such as a whitening agent from the viewpoint of safety.
　サルオガセ属地衣類は、ジフラクタ酸やウスニン酸を含有することが知られている（例えば、Phytochemistry，1987，vol．26，No．12，p．3181-3185；Bull．Kochi．Gakuen　College.，1992，vol．23，p．687(9)-697(19)参照）。 Salogase lichen is known to contain difractic acid and usnic acid (for example, Phytochemistry, 1987, vol. 26, No. 12, p. 3181-3185; Bull. Kochi. Gakuen College., 1992, vol.23, p.687 (9) -697 (19)).
　そこで、本発明者らは、美白剤などの皮膚外用剤に好適に用いることができる、ウスニン酸の含有量を低減した、サルオガセ属地衣類の抽出条件について検討を行った。その結果、アルコール含有率の高いアルコール水溶液を抽出溶媒として用いてサルオガセ属地衣類を抽出するとウスニン酸の濃度が上昇するのに対し、アルコール含有率が一定範囲のアルコール水溶液を抽出溶媒として用いてサルオガセ属地衣類を抽出するとウスニン酸の濃度が低減できることを見出した。
　さらに、アルコール含有率が一定範囲のアルコール水溶液を抽出溶媒として用いて抽出したサルオガセ属地衣類抽出物は、エンドセリンの作用によって引き起こされるメラノサイト内のカルシウム（イオン）濃度上昇の抑制効果を有することを見出した。 Therefore, the present inventors have studied the extraction conditions of Sarugagase lichen with reduced usnic acid content, which can be suitably used for skin external preparations such as whitening agents. As a result, the concentration of usnic acid increases when an aqueous solution of alcohol with high alcohol content is used as the extraction solvent, while the concentration of usnic acid increases, while the aqueous solution of alcohol with a certain range of alcohol content is used as the extraction solvent. It was found that the concentration of usnic acid can be reduced by extracting clothing.
Furthermore, it was found that an extract of Lichenaceae lichen extracted using an aqueous alcohol solution with a certain alcohol content range as an extraction solvent has an inhibitory effect on the increase in calcium (ion) concentration in melanocytes caused by the action of endothelin. .
　さらに、本発明者らは、美白剤などの皮膚外用剤に好適に用いることができ、毒性を有するウスニン酸の含有量を低減した、サルオガセ属地衣類抽出物又はエンドセリン作用抑制剤が得られる製造方法について検討を行った。その結果、アルコール含有率が一定範囲のアルコール水溶液を抽出溶媒として用いてサルオガセ属地衣類を抽出することにより、ジフラクタ酸濃度が有意に上昇し、肝毒性物質であるウスニン酸濃度が有意に低下し、美白剤などの皮膚外用剤に好適に用いることができる、サルオガセ属地衣類抽出物又はエンドセリン作用抑制剤が得られるとの知見を得た。 Furthermore, the present inventors can use the Sargogase lichen extract or endothelin action inhibitor which can be used suitably for skin external preparations such as whitening agents and has a reduced content of toxic usnic acid. Was examined. As a result, by extracting Saruogase lichen using an aqueous alcohol solution with a certain range of alcohol content as an extraction solvent, the concentration of difractic acid is significantly increased, and the concentration of usnic acid, a hepatotoxic substance, is significantly decreased. The inventor has obtained knowledge that a Sargogase lichen extract or endothelin action inhibitor can be suitably used for a skin external preparation such as a whitening agent.
　さらに、本発明者らは、美白剤などの皮膚外用剤に好適に用いることができ、毒性を有するウスニン酸の含有量を選択的に低減した、サルオガセ属地衣類抽出物又はエンドセリン作用抑制剤が得られる製造方法について検討を行った。その結果、アルコール含有率が７０体積％を超えるアルコール水溶液を抽出溶媒として用い調製したサルオガセ属地衣類の抽出物について、低温保存処理や加水処理を行うことにより、肝毒性物質であるウスニン酸濃度が有意に低下し、美白剤などの皮膚外用剤に好適に用いることができるサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤が得られるとの知見を得た。 Furthermore, the present inventors can obtain a Sargogase lichen extract or endothelin action inhibitor that can be suitably used for a topical skin preparation such as a whitening agent and selectively reduces the content of toxic usnic acid. The production method to be used was examined. As a result, the concentration of usnic acid, which is a hepatotoxic substance, is significant by performing low-temperature storage treatment and hydration treatment on the extract of Lichenaceae liquor prepared using an aqueous alcohol solution with an alcohol content exceeding 70% by volume as an extraction solvent. It was found that a Sargogase lichen extract or endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent is obtained.
　本発明はこれらの知見に基づいて完成するに至ったものである。 The present invention has been completed based on these findings.
　本発明、特に本発明の第１の実施態様の製造方法により得られるサルオガセ属地衣類抽出物は、肝毒性物質であるウスニン酸の含有量が低減されている。したがって、本発明、特に本発明の第１の実施態様の製造方法は、エンドセリン作用抑制剤、メラニン生成阻害剤、美白剤などの皮膚外用剤に好適に用いることができるサルオガセ属地衣類抽出物を製造することができる。 The present invention, in particular, the Sargogus genus lichen extract obtained by the production method of the first embodiment of the present invention has a reduced content of usnic acid, which is a hepatotoxic substance. Therefore, the production method of the present invention, particularly the first embodiment of the present invention, produces an extract of Sarugagase lichen that can be suitably used for external skin preparations such as endothelin action inhibitors, melanin production inhibitors, and whitening agents. can do.
　本発明、特に本発明の第２の実施態様の製造方法により得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤は、エンドセリン作用抑制剤の有効成分であるジフラクタ酸を高含有するとともに、肝毒性物質であるウスニン酸の含有量が低減されている。したがって、本発明、特に本発明の第２の実施態様の製造方法は、美白剤などの皮膚外用剤に好適に用いることができるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤を製造することができる。 The present invention, and in particular, the extract of Lichenaceae lichen and endothelin action inhibitor obtained by the production method of the second embodiment of the invention contains a high content of difractic acid, which is an active ingredient of endothelin action inhibitor, and is a hepatotoxic substance. The content of usnic acid is reduced. Therefore, the production method of the present invention, particularly the second embodiment of the present invention, can produce a Salomagus lichen extract and an endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent.
　本発明、特に本発明の第３の実施態様の製造方法により得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤は、肝毒性物質であるウスニン酸の含有量が低減されている。したがって、本発明、特に本発明の第３の実施態様の製造方法は、美白剤などの皮膚外用剤に好適に用いることができるサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤を製造することができる。 In the present invention, particularly in the Sargoguce lichen extract and endothelin action inhibitor obtained by the production method of the third embodiment of the present invention, the content of usnic acid, which is a hepatotoxic substance, is reduced. Therefore, the production method of the present invention, particularly the third embodiment of the present invention, can produce a Sargogase lichen extract or endothelin action inhibitor that can be suitably used for a skin external preparation such as a whitening agent.
　本発明の上記及び他の特徴及び利点は、下記の記載からより明らかになるであろう。 The above and other features and advantages of the present invention will become more apparent from the following description.
　本発明の第１の実施態様のウスニン酸低含有サルオガセ属地衣類抽出物の製造方法において、アルコール含有率が８０体積％以下のアルコール水溶液を用いてサルオガセ属地衣類を抽出する。
　当該方法により、肝毒性物質であるウスニン酸の含有量が低いサルオガセ属地衣類抽出物が得られる。このようにして得られるサルオガセ属地衣類抽出物は安全性に優れ、エンドセリン作用抑制効果、メラニン生成阻害効果、美白効果などを発揮し得る医薬品、医薬部外品、化粧料等に好適に用いることができる。 In the method for producing an extract of low usnin acid sorghum lichen according to the first embodiment of the present invention, the sorghum lichen is extracted using an aqueous alcohol solution having an alcohol content of 80% by volume or less.
According to this method, an extract of Lichenaceae lichen having a low content of usnic acid, which is a hepatotoxic substance, can be obtained. The extract of the genus Sarhogase obtained in this way is excellent in safety and should be suitably used for pharmaceuticals, quasi-drugs, cosmetics, etc. that can exhibit endothelin action inhibitory effect, melanin production inhibitory effect, whitening effect, etc. it can.
　本明細書において、「ウスニン酸低含有サルオガセ属地衣類抽出物」とは、同条件で抽出を行った場合、本発明の第１の実施態様で規定する抽出溶媒を用いて得られるサルオガセ属地衣類抽出物中のウスニン酸含有量が、本発明の第１の実施態様で規定する抽出溶媒よりもアルコール含有率の高い抽出溶媒を用いて得られる抽出物中のウスニン酸含有量よりも少ないことを意味し、同一の溶媒種を用い同条件で抽出した場合にアルコール含有率を変化させたときの抽出物中のウスニン酸濃度を比較したものである。 In the present specification, the term “usunic acid low-content Salogase lichen extract” refers to the extraction of Salogase lichen obtained using the extraction solvent defined in the first embodiment of the present invention when extraction is performed under the same conditions. It means that the usnic acid content in the product is less than the usnic acid content in the extract obtained using the extraction solvent having a higher alcohol content than the extraction solvent defined in the first embodiment of the present invention. In addition, the concentration of usnic acid in the extracts when the alcohol content is changed when the same solvent species is extracted under the same conditions is compared.
　本発明の第２の実施態様のジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法及びエンドセリン作用抑制剤の製造方法において、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いてサルオガセ属地衣類を抽出する。
　当該方法により、エンドセリン作用抑制効果を有するジフラクタ酸を高濃度で含有し、肝毒性物質であるウスニン酸の含有量が低いサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤が得られる。このようにして得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤は、ジフラクタ酸の美白効果を発揮し得る医薬品、医薬部外品、化粧料等に好適に用いることができる。 In the method for producing a high extract of difractate and low content of usnic acid and a method for producing endothelin action inhibitor according to the second embodiment of the present invention, an alcohol having an alcohol content of 30% by volume to 80% by volume Extract Sarogase lichen using aqueous solution.
According to this method, an extract of Salgocase genus lichen or endothelin action inhibitor containing difractic acid having an endothelin action inhibitory effect at a high concentration and a low content of usnic acid, which is a hepatotoxic substance, is obtained. The Saruogase lichen extract and endothelin action inhibitor thus obtained can be suitably used for pharmaceuticals, quasi-drugs, cosmetics and the like that can exert the whitening effect of difractic acid.
　本明細書において、「ジフラクタ酸高含有サルオガセ属地衣類抽出物」とは、同条件で抽出を行った場合、本発明の第２の実施態様で規定する抽出溶媒を用いて得られるサルオガセ属地衣類抽出物中のジフラクタ酸含有量が、本発明で規定する抽出溶媒よりもアルコール含有率の低い抽出溶媒を用いて得られる抽出物中のジフラクタ酸含有量よりも多いことを意味し、同一の溶媒種を用い同条件で抽出した場合にアルコール含有率を変化させたときの抽出物中のジフラクタ酸濃度を比較したものである。
　また、「ウスニン酸低含有サルオガセ属地衣類抽出物」とは、同条件で抽出を行った場合、本発明の第２の実施態様で規定する抽出溶媒を用いて得られるサルオガセ属地衣類抽出物中のウスニン酸含有量が、本発明で規定する抽出溶媒よりもアルコール含有率の高い抽出溶媒を用いて得られる抽出物中のウスニン酸含有量よりも少ないことを意味し、同一の溶媒種を用い同条件で抽出した場合にアルコール含有率を変化させたときの抽出物中のウスニン酸濃度を比較したものである。
　なお、後述の実施例で示すように、本発明の第２の実施態様で規定する抽出溶媒を用いて得られるサルオガセ属地衣類抽出物中のジフラクタ酸含有量が、本発明の第２の実施態様で規定する抽出溶媒よりもアルコール含有率の高い抽出溶媒を用いて得られる抽出物中のジフラクタ酸含有量よりも低い場合であっても、ウスニン酸の含有率を比較した場合、その含有率が、本発明の第２の実施態様で規定する抽出溶媒を用いて得られる抽出物の方が低い場合には、本発明の第２の実施態様で規定する抽出溶媒を用いて得られるサルオガセ属地衣類抽出物は本明細書における「ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物」に含むものとする。 In the present specification, “the extract of the genus Sarcogella having a high content of difructaic acid” refers to the extraction of the genus Lichenaceus obtained using the extraction solvent defined in the second embodiment of the present invention when extraction is performed under the same conditions. Means that the content of difractic acid in the product is higher than the content of difractic acid in the extract obtained by using an extraction solvent having an alcohol content lower than that of the extraction solvent defined in the present invention. Is a comparison of the concentration of difractic acid in the extract when the alcohol content is changed when extracted under the same conditions.
In addition, “usunic acid low content Salogase lichen extract” in the extract of Salogase lichen obtained using the extraction solvent defined in the second embodiment of the present invention when extraction is performed under the same conditions. This means that the usnic acid content is lower than the usnic acid content in the extract obtained using an extraction solvent having a higher alcohol content than the extraction solvent specified in the present invention. This is a comparison of the usnic acid concentration in the extract when the alcohol content is changed when extracted under the conditions.
In addition, as shown in the below-mentioned Examples, the content of difractic acid in the extract of Lichenacea lichen obtained using the extraction solvent defined in the second embodiment of the present invention is the second embodiment of the present invention. Even if it is lower than the content of difractic acid in the extract obtained by using an extraction solvent having a higher alcohol content than the extraction solvent specified in 1. In the case where the extract obtained using the extraction solvent defined in the second embodiment of the present invention is lower in the extract, the Lichenaceae lichen obtained using the extraction solvent defined in the second embodiment of the present invention The extract is to be included in the “Dioglucate-rich and usnic acid-rich Salogase lichen extract” in the present specification.
　本明細書において、特に断らない限り、本発明の第３の実施態様とは下記第４及び第５の実施態様のすべてを包含する意味である。 In this specification, unless otherwise specified, the third embodiment of the present invention is meant to include all of the following fourth and fifth embodiments.
　本発明の第４の実施態様は、サルオガセ属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理（本明細書において、「ウスニン酸の選択的低減処理」ともいう）を行う抽出物の製造方法であって、前記処理後のウスニン酸の残存率が５０％未満である、ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法である。当該方法により、肝毒性物質であるウスニン酸の含有量が低いサルオガセ属地衣類抽出物が得られる。
　また、本発明の第５の実施態様は、サルオガセ属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う、ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物又はエンドセリン作用抑制剤の製造方法である。当該方法により、エンドセリン作用抑制効果を有するジフラクタ酸を高濃度で含有し、肝毒性物質であるウスニン酸の含有量が低い、サルオガセ属地衣類抽出物又はエンドセリン作用抑制剤が得られる。
　このようにして得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤は、美白効果などを発揮し得る医薬品、医薬部外品、化粧料等に好適に用いることができる。 According to a fourth embodiment of the present invention, a sorghum lichen is extracted using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and hydration treatment. A method for producing an extract that performs one or more selected treatments (also referred to herein as “selective reduction treatment of usnic acid”), wherein the residual ratio of usnic acid after the treatment is less than 50% It is a manufacturing method of a certain usnusic acid low content Salogase lichen extract. According to this method, an extract of Lichenaceae lichen having a low content of usnic acid, which is a hepatotoxic substance, can be obtained.
Further, the fifth embodiment of the present invention is a method for extracting Sarcogum lichen using an aqueous alcohol solution having an alcohol content of more than 70% by volume, and comprising a low-temperature storage treatment and a hydration treatment for the obtained extract. It is a method for producing an extract containing a high content of difractic acid and a low content of usnic acid, or an endothelin action inhibitor, which performs at least one treatment selected from the group. According to this method, an extract of Saruogase lichen or endothelin action that contains difractic acid having an endothelin action inhibitory effect at a high concentration and a low content of usnic acid, which is a hepatotoxic substance, is obtained.
The Saruogase lichen extract and endothelin action inhibitor thus obtained can be suitably used for pharmaceuticals, quasi-drugs, cosmetics and the like that can exhibit a whitening effect.
　本明細書において、「ウスニン酸低含有サルオガセ属地衣類抽出物」とは、本発明の第４及び第５の実施態様で規定する抽出溶媒を用いて抽出したサルオガセ属地衣類抽出物に含まれるウスニン酸含有量と比較して、当該抽出物に対してさらに低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行って得られるサルオガセ属地衣類抽出物中のウスニン酸の含有量の割合が低減されていることを意味する。
　また、本発明の第４の実施態様における「前記処理後のウスニン酸の残存率が５０％未満である」とは、前記ウスニン酸の選択的低減処理後の抽出物中のウスニン酸含有量を、希釈分を考慮し、ウスニン酸の選択的低減処理前の含有量に換算した場合、ウスニン酸の選択的低減処理後の抽出物中のウスニン酸含有量がウスニン酸の選択的低減処理処理前の抽出物中のウスニン酸含有量の５０％未満に低減されていることを意味する。 In the present specification, the “usunic acid low-content Sarhogase lichen extract” refers to usnic acid contained in the Sarusose lichen extract extracted using the extraction solvent defined in the fourth and fifth embodiments of the present invention. The proportion of the content of usnic acid in the extract of Lichenaceae lichen obtained by performing one or more treatments selected from the group consisting of low-temperature storage treatment and hydration treatment on the extract as compared with the content Is reduced.
Further, in the fourth embodiment of the present invention, “the residual ratio of usnic acid after the treatment is less than 50%” means that the usnic acid content in the extract after the selective reduction treatment of usnic acid is In consideration of dilution, when converted to the content before selective reduction treatment of usnic acid, the content of usnic acid in the extract after selective reduction treatment of usnic acid is before the selective reduction treatment of usnic acid. Is reduced to less than 50% of the usnic acid content in the extract.
　本発明の第５の実施態様における「ジフラクタ酸高含有サルオガセ属地衣類抽出物」とは、同条件で抽出を行った場合、本発明の第５の実施態様で規定する抽出溶媒を用いて抽出したサルオガセ属地衣類抽出物中のジフラクタ酸含有量が、本発明の第５の実施態様で規定する抽出溶媒よりもアルコール含有率の低い抽出溶媒を用いて抽出した抽出物中のジフラクタ酸含有量よりも多いことを意味し、同一の溶媒種を用いて同条件で抽出した場合にアルコール含有率を変化させたときの抽出物中のジフラクタ濃度を比較したものである。 In the fifth embodiment of the present invention, the “difractate-rich Salogase lichen extract” is extracted using the extraction solvent defined in the fifth embodiment of the present invention when extraction is performed under the same conditions. The content of difractic acid in the extract of Lichenaceae is higher than the content of difractic acid in the extract extracted using an extraction solvent having a lower alcohol content than the extraction solvent defined in the fifth embodiment of the present invention. This means that the concentration of difractor in the extract when the alcohol content is changed when extracted under the same conditions using the same solvent species is compared.
　本発明の第２及び第５の実施態様の製造方法により得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤において、ジフラクタ酸は塩の形態で存在してもよい。ジフラクタ酸の塩としては適宜選択することができ、例えば、リチウム塩、ナトリウム塩、カリウム塩等のアルカリ金属塩、カルシウム塩、マグネシウム塩等のアルカリ土類金属塩、トリメチルアミン、トリエチルアミン等のアルキルアミン塩及び４級アンモニウム塩、トリエタノールアミン、ジエタノールアミン、モノエタノールアミン等のアルカノールアミン塩、又はリジン、ヒスチジン、アルギニン等のアミノ酸塩などが挙げられ、好ましくは、アルカリ金属塩、アルカノールアミン塩、アミノ酸塩である。 In the Sargogus genus lichen extract and the endothelin action inhibitor obtained by the production methods of the second and fifth embodiments of the present invention, difractic acid may be present in the form of a salt. The salt of difractic acid can be appropriately selected. For example, alkali metal salts such as lithium salt, sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, and alkylamine salts such as trimethylamine and triethylamine. And alkanolamine salts such as quaternary ammonium salts, triethanolamine, diethanolamine, and monoethanolamine, or amino acid salts such as lysine, histidine, and arginine. Preferred are alkali metal salts, alkanolamine salts, and amino acid salts. is there.
　本発明で用いるサルオガセ属地衣類は、ウメノキゴケ（Parmeliaceae）科の地衣類である。本発明では、天然に存在するサルオガセ属地衣類を採取して用いてもよいし、市販品を入手して用いてもよい。本発明に用いられるサルオガセ属地衣類としては適宜選択することができ、ナガサルオガセ（Usnea　longissima）、ヨコワサルオガセ（Usnea　diffracta　Vain.）、フジサルオガセ（Usnea　trichodeoides）、ニセフジサルオガセ（Usnea　pseudomontis-fuji）、クシノハサルオガセ（Usnea　pectinata）などが挙げられる。本発明で用いる前記サルオガセ属地衣類としては、ナガサルオガセ、ヨコワサルオガセ、フジサルオガセ、ニセフジサルオガセ及びクシノハサルオガセからなる群より選ばれる１種又は２種以上が好ましく、ナガサルオガセ及びヨコワサルオガセからなる群より選ばれる１種又は２種がより好ましく、ナガサルオガセ又はヨコワサルオガセがさらに好ましい。
　本発明において、前記サルオガセ属地衣類の全ての任意の部位（全体、地衣体、仮根、シリア、臍状体（さいじょうたい）等）を用いることができ、各部位を複数組み合わせて用いてもよい。また地位培養細胞（菌細胞、藻細胞、又は両者の混在した未分化共生培養組織）を用いることもできる。本発明において、サルオガセ属地衣類を抽出するには、地衣体を用いるのが好ましい。
　本発明に用いるサルオガセ属地衣類は、天然に存在するそのままの状態でもよいが、抽出効率を高めるために、乾燥、凍結乾燥、細断、粉砕などの処理を施したものを用いるのが好ましい。 The Lichenaceae lichen used in the present invention is a lichen belonging to the family Parmeliaceae. In the present invention, naturally-occurring Salogase lichens may be collected and used, or commercially available products may be obtained and used. As Usnea territorial clothing for use in the present invention can be appropriately selected, Naga Usnea (Usnea longissima), Yokowa Usnea (Usnea diffracta Vain.), Fuji Usnea (Usnea trichodeoides), false Fuji Usnea (Usnea pseudomontis-fuji), Examples include Usnea pectinata . As the lichen genus lichen used in the present invention, one or two or more selected from the group consisting of Nagasaru Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefujisaru Ogase and Kushinohasa Ogase are preferred, and the group consisting of Nagasara Ogase and Yokosawa Ogase 1 type or 2 types selected more are more preferable, and Nagasaru Ogase or Yokosawa Ogase is still more preferable.
In the present invention, any arbitrary part (the whole body, lichen body, temporary root, Syria, umbilical body, etc.) of the genus Ligoceae can be used, and a plurality of each part may be used in combination. . In addition, standing culture cells (fungal cells, algal cells, or an undifferentiated symbiotic cultured tissue in which both are mixed) can also be used. In the present invention, it is preferable to use a lichen body in order to extract the Sarhogase lichen.
The Sarhogase lichen used in the present invention may be in a naturally existing state, but in order to increase the extraction efficiency, it is preferable to use one subjected to treatment such as drying, freeze-drying, shredding, and pulverization.
　本発明の第１の実施態様の製造方法において、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いる。本発明の第２の実施態様の製造方法において、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いる。本発明の第３の実施態様の製造方法においては、アルコール含有率が７０体積％を超えるアルコール水溶液を抽出溶媒として用いる。
　本発明において抽出溶媒として用いるアルコール水溶液としては、メタノール、エタノール、イソプロピルアルコール、ブタノール等の低級アルコール類（好ましくは、炭素数１～４の低級アルコール類）の水溶液、1,2-ブタンジオール、1,3-ブタンジオール、1,4-ブタンジオール、2,3-ブタンジオール、エチレングリコール、プロピレングリコール、1,3-ブチレングリコール等の多価アルコール類の水溶液が挙げられる。本発明では、これらの溶媒を単独で用いても、２種以上を組み合わせて用いてもよい。本発明の第１及び第２の実施態様においては、メタノール、エタノール、イソプロピルアルコール、1,2-ブタンジオール、1,3-ブタンジオール、1,4-ブタンジオール、2,3-ブタンジオール等の水溶液を用いるのが好ましく、メタノール、エタノール、1,2-ブタンジオール、1,3-ブタンジオール、1,4-ブタンジオール又は2,3-ブタンジオールの水溶液を用いるのがより好ましく、エタノール水溶液又は1,3-ブタンジオール水溶液がさらに好ましい。本発明の第３の実施態様においては、メタノール、エタノール、イソプロピルアルコール等の低級アルコール類の水溶液を用いるのが好ましく、メタノール又はエタノールの水溶液を用いるのがより好ましく、エタノール水溶液がさらに好ましい。
　本発明の第１の実施態様において抽出溶媒として用いるアルコール水溶液のアルコール含有率は７０体積％以下が好ましく、６０体積％以下がより好ましい。
　本発明の第２の実施態様において抽出溶媒として用いるアルコール水溶液のアルコール含有率は、４０体積％以上が好ましく、７０体積％以下が好ましく、６０体積％以下がより好ましい。また、３０体積％以上７０体積％以下が好ましく、４０体積％以上７０体積％以下がさらに好ましく、４０体積％以上６０体積％以下が特に好ましい。
　本発明の第３の実施態様において抽出溶媒として用いるアルコール水溶液のアルコール含有率は８０体積％を超えるのが好ましく、９０体積％以上がより好ましく、９５体積％以上がさらに好ましい。 In the production method of the first embodiment of the present invention, an alcohol aqueous solution having an alcohol content of 80% by volume or less is used as an extraction solvent. In the production method of the second embodiment of the present invention, an alcohol aqueous solution having an alcohol content of 30% by volume to 80% by volume is used as the extraction solvent. In the production method of the third embodiment of the present invention, an aqueous alcohol solution having an alcohol content of more than 70% by volume is used as the extraction solvent.
Examples of the aqueous alcohol solution used as the extraction solvent in the present invention include aqueous solutions of lower alcohols (preferably lower alcohols having 1 to 4 carbon atoms) such as methanol, ethanol, isopropyl alcohol, and butanol, 1,2-butanediol, 1 , 3-butanediol, 1,4-butanediol, 2,3-butanediol, aqueous solutions of polyhydric alcohols such as ethylene glycol, propylene glycol, and 1,3-butylene glycol. In the present invention, these solvents may be used alone or in combination of two or more. In the first and second embodiments of the present invention, methanol, ethanol, isopropyl alcohol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 2,3-butanediol, etc. It is preferable to use an aqueous solution, and it is more preferable to use an aqueous solution of methanol, ethanol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, or 2,3-butanediol. A 1,3-butanediol aqueous solution is more preferred. In the third embodiment of the present invention, it is preferable to use an aqueous solution of lower alcohols such as methanol, ethanol, isopropyl alcohol, more preferably an aqueous solution of methanol or ethanol, and even more preferably an aqueous ethanol solution.
In the first embodiment of the present invention, the alcohol content of the alcohol aqueous solution used as the extraction solvent is preferably 70% by volume or less, and more preferably 60% by volume or less.
The alcohol content of the aqueous alcohol solution used as the extraction solvent in the second embodiment of the present invention is preferably 40% by volume or more, preferably 70% by volume or less, and more preferably 60% by volume or less. Moreover, 30 volume% or more and 70 volume% or less are preferable, 40 volume% or more and 70 volume% or less are more preferable, and 40 volume% or more and 60 volume% or less are especially preferable.
The alcohol content of the aqueous alcohol solution used as the extraction solvent in the third embodiment of the present invention is preferably more than 80% by volume, more preferably 90% by volume or more, and still more preferably 95% by volume or more.
　サルオガセ属地衣類の抽出条件については、使用する抽出溶媒によって適宜設定できる。例えば、抽出温度は、０℃以上が好ましく、５℃以上がより好ましく、５０℃以下が好ましく、４０℃以下がより好ましく、２０℃以下がさらに好ましい。あるいは、０～５０℃が好ましく、５～４０℃がより好ましく、５～２０℃がさらに好ましい。抽出時間は、１時間以上が好ましく、１日以上がより好ましく、２０日以下が好ましく、７日以下がより好ましい。あるいは、１時間～２０日間が好ましく、１～７日間がより好ましい。抽出効率を上げるため、抽出の際に攪拌を行ったり、溶媒中でホモジナイズ処理を行ってもよい。なお、抽出溶媒の使用量は適宜設定することができ、抽出原料のサルオガセ属地衣類に対し質量比で１倍量以上であることが好ましく、５倍量以上であることがより好ましく、５０倍量以下であることが好ましく、３０倍量以下であることがより好ましい。あるいは、１～５０倍量であることが好ましく、５～３０倍量であることがより好ましい。 About the extraction conditions of Saruogase lichen, it can set suitably with the extraction solvent to be used. For example, the extraction temperature is preferably 0 ° C. or higher, more preferably 5 ° C. or higher, preferably 50 ° C. or lower, more preferably 40 ° C. or lower, and further preferably 20 ° C. or lower. Alternatively, 0 to 50 ° C is preferable, 5 to 40 ° C is more preferable, and 5 to 20 ° C is further preferable. The extraction time is preferably 1 hour or longer, more preferably 1 day or longer, preferably 20 days or shorter, more preferably 7 days or shorter. Alternatively, it is preferably 1 hour to 20 days, more preferably 1 to 7 days. In order to increase extraction efficiency, stirring may be performed during extraction, or homogenization treatment may be performed in a solvent. In addition, the usage-amount of an extraction solvent can be set suitably, It is preferable that it is 1 time amount or more by mass ratio with respect to the Salogase lichen of an extraction raw material, It is more preferable that it is 5 times amount or more, 50 times amount Or less, more preferably 30 times or less. Alternatively, the amount is preferably 1 to 50 times, and more preferably 5 to 30 times.
　本発明の第１及び第２の実施態様において、上記溶媒で抽出して得られた抽出物はそのまま使用してもよいが、必要に応じて更に適当な分離手段、例えばゲル濾過、クロマトグラフィー、精密蒸留等により活性の高い画分を分画してもよい。あるいは、得られた抽出物の溶媒を別の溶媒（例えば、アルコール水溶液）に置換してもよい。本発明の製造方法により得られる抽出物には、このようにして得られた各種抽出物、その希釈液、その濃縮液、その精製物又はそれらの乾燥末等が包含される。
　本発明の第３の実施態様において、上記溶媒で抽出して得られた抽出物はそのままウスニン酸の選択的低減処理に使用してもよいが、必要に応じて更に適当な分離手段、例えばゲル濾過、クロマトグラフィー、精密蒸留等により活性の高い画分を分画してもよい。あるいは、得られた抽出物の溶媒を別の溶媒（例えば、アルコール水溶液）に置換してもよい。本発明の製造方法に用いられる溶媒抽出後の抽出物には、このようにして得られた各種抽出物、その希釈液、その濃縮液、その精製物又はそれらの乾燥末等が包含される。 In the first and second embodiments of the present invention, the extract obtained by extraction with the solvent may be used as it is, but if necessary, further suitable separation means such as gel filtration, chromatography, A highly active fraction may be fractionated by precision distillation or the like. Alternatively, the solvent of the obtained extract may be replaced with another solvent (for example, an aqueous alcohol solution). The extract obtained by the production method of the present invention includes the various extracts thus obtained, a diluted solution thereof, a concentrated solution thereof, a purified product thereof or a dry powder thereof.
In the third embodiment of the present invention, the extract obtained by extraction with the solvent may be used as it is for selective reduction of usnic acid, but if necessary, further suitable separation means such as a gel is used. A highly active fraction may be fractionated by filtration, chromatography, precision distillation or the like. Alternatively, the solvent of the obtained extract may be replaced with another solvent (for example, an aqueous alcohol solution). The extract after solvent extraction used in the production method of the present invention includes the various extracts thus obtained, a diluted solution thereof, a concentrated solution thereof, a purified product thereof or a dry powder thereof.
　本発明の第１の実施態様において、抽出溶媒としてアルコール含有率が８０体積％以下のアルコール水溶液を用いて得られるサルオガセ属地衣類抽出物を皮膚外用剤に用いることを考慮して、サルオガセ属地衣類抽出物に対してさらなるウスニン酸の低減処理を施すことが好ましい。
　ウスニン酸の低減処理としては任意の方法を選択することができ、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理、加熱処理などがあげられる。本発明では、これらの処理工程を単独で用いてもよいし、２種以上を組合わせて用いてもよく、これらの処理工程を繰返し行ってもよい。 In the first embodiment of the present invention, taking into account the use of the extract of Lichenaceae lichen obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent for the topical skin preparation, It is preferable to further reduce the usnic acid to the product.
Any method can be selected as the usnic acid reduction treatment, and examples thereof include low-temperature storage treatment, water addition treatment, activated carbon treatment, hexane washing treatment, and heat treatment. In the present invention, these treatment steps may be used alone or in combination of two or more, and these treatment steps may be repeated.
　本発明の第２の実施態様において、抽出溶媒としてアルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤を皮膚外用剤に用いることを考慮して、サルオガセ属地衣類抽出物に対してさらにウスニン酸の選択的低減処理を施すことが好ましい。
　ウスニン酸の選択的低減処理としては任意の方法を選択することができ、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理、加熱処理などが挙げられる。本発明では、これらの処理工程を単独で用いてもよいし、２種以上を組合わせて用いてもよく、これらの処理工程を繰返し行ってもよい。
　なお、ウスニン酸の選択的低減処理によりサルオガセ属地衣類抽出物中のジフラクタ酸含有量が低下しても、ジフラクタ酸含有量の低下率と比較してウスニン酸含有量の低下率が十分に高ければよい。 In the second embodiment of the present invention, an extract of Salogase lichen and an endothelin action inhibitor obtained using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is used as a skin external preparation. In consideration of the above, it is preferable to further perform a selective reduction treatment of usnic acid on the Sarhogase lichen extract.
As the selective reduction treatment of usnic acid, any method can be selected, and examples thereof include low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment, and heat treatment. In the present invention, these treatment steps may be used alone or in combination of two or more, and these treatment steps may be repeated.
Even if the difractic acid content in the extract of Lichenaceae is reduced by the selective reduction treatment of usnic acid, the decrease rate of usnic acid content is sufficiently high compared to the decrease rate of difractic acid content. Good.
　本発明の第３の実施態様では、アルコール含有率が７０体積％を超えるアルコール水溶液を用いてサルオガセ属地衣類を抽出し、得られた抽出物を低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う。
　本発明では、前記ウスニン酸の選択的低減処理を単独で用いてもよいし、２種以上を組合わせて用いてもよく、これらの処理工程を繰返し行ってもよい。また、本発明の第４の実施態様において、サルオガセ属地衣類抽出物のウスニン酸の残存率を５０％未満とするため、前記ウスニン酸の選択的低減処理の順序、回数などを適宜設計・変更することができる。 In the third embodiment of the present invention, Salgogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, and the obtained extract is selected from the group consisting of low-temperature storage treatment and water treatment. Process more than seeds.
In the present invention, the selective reduction treatment of usnic acid may be used alone, or two or more may be used in combination, and these treatment steps may be repeated. Further, in the fourth embodiment of the present invention, the order and number of times of the selective reduction of usnic acid are appropriately designed and changed in order to make the residual rate of usnic acid in the extract of Lichenaceae liquor less than 50%. be able to.
　ウスニン酸の（選択的）低減処理としての低温保存について説明する。
　ウスニン酸は結晶性が高く、水溶液に対する溶解性が低く、低温では析出しやすい。そのため、本発明で規定する抽出溶媒を用いて抽出したサルオガセ属地衣類抽出物を低温保存すると、ウスニン酸が溶けにくくなりウスニン酸の結晶が析出する。このようにして析出したウスニン酸を常法により除去することで、サルオガセ属地衣類抽出物からのウスニン酸の（選択的）低減が可能となる。
　サルオガセ属地衣類抽出物の保存温度としては－１０℃以上が好ましく、－５℃以上がより好ましく、５℃以下が好ましい。あるいは、－１０～５℃が好ましく、－５～５℃がより好ましい。また、本発明の第１及び第２の実施態様において、サルオガセ属地衣類抽出物の保存時間としては１時間以上が好ましく、６時間以上がより好ましく、１４日以下が好ましく、５日以下がより好ましい。あるいは、１時間～１４日間が好ましく、６時間～５日間がより好ましい。また、本発明の第３の実施態様において、サルオガセ属地衣類抽出物の保存時間としては１時間以上が好ましく、３時間以上がより好ましく、６時間以上がさらに好ましく、４０日以下が好ましく、１４日以下がより好ましく、７日以下がより好ましい。あるいは、１時間～４０日間が好ましく、３時間～１４日間がより好ましく、６時間～７日間がさらに好ましい。 The low temperature storage as a (selective) reduction process of usnic acid will be described.
Usnic acid has high crystallinity, low solubility in aqueous solutions, and tends to precipitate at low temperatures. For this reason, when the Lichenaceae lichen extract extracted using the extraction solvent defined in the present invention is stored at a low temperature, usnic acid is hardly dissolved and crystals of usnic acid are precipitated. By removing the precipitated usnic acid by a conventional method, it becomes possible to (selectively) reduce the usnic acid from the Lichenaceae lichen extract.
The storage temperature of the extract of Lichenaceae is preferably −10 ° C. or higher, more preferably −5 ° C. or higher, and preferably 5 ° C. or lower. Alternatively, −10 to 5 ° C. is preferable, and −5 to 5 ° C. is more preferable. Moreover, in the 1st and 2nd embodiment of this invention, as a preservation | save time of a Sarhogase lichen extract, 1 hour or more is preferable, 6 hours or more are more preferable, 14 days or less are preferable, and 5 days or less are more preferable. . Alternatively, it is preferably 1 hour to 14 days, more preferably 6 hours to 5 days. In addition, in the third embodiment of the present invention, the storage time of the extract of Lichenaceae liquor is preferably 1 hour or longer, more preferably 3 hours or longer, further preferably 6 hours or longer, preferably 40 days or shorter, 14 days The following is more preferable, and 7 days or less is more preferable. Alternatively, it is preferably 1 hour to 40 days, more preferably 3 hours to 14 days, and even more preferably 6 hours to 7 days.
　ウスニン酸の（選択的）低減処理としての加水処理について説明する。
　ウスニン酸は結晶性が高く、水溶液に対する溶解性が低い。そのため、本発明で規定する抽出溶媒を用いて抽出したサルオガセ属地衣類抽出物を含水率の高い溶液中で保存すると、ウスニン酸が溶けにくくなりウスニン酸の結晶が析出する。このようにして析出したウスニン酸を常法により除去することで、サルオガセ属地衣類抽出物からのウスニン酸の（選択的）低減が可能となる。
　本発明の第１の実施態様において、サルオガセ属地衣類抽出物に対する加水量としては適宜設定することができる。本発明の第２の実施態様において、サルオガセ属地衣類抽出物に対する加水量としては、加水後にジフラクタ酸が析出等によりロスしない量の範囲で適宜設定することができる。具体的には、加水後のアルコール含有率が２５体積％以上となるように加水するのが好ましく、３０体積％以上となるように加水するのがさらに好ましく、４５体積％以下となるように加水するのが好ましく、４０体積％以下となるように加水するのがさらに好ましい。あるいは、加水後のアルコール含有率が２５～４５体積％となるように加水するのが好ましく、３０～４０体積％となるように加水するのがさらに好ましい。
　本発明の第３の実施態様において、サルオガセ属地衣類抽出物に対する加水量としては適宜設定することができ、加水後のアルコール含有率が２５体積％以上となるように加水するのが好ましく、３０体積％以上となるように加水するのがさらに好ましく、５０体積％以下となるように加水するのが好ましく、４５体積％以下となるように加水するのがさらに好ましい。あるいは、加水後のアルコール含有率が２５～５０体積％となるように加水するのが好ましく、３０～４５体積％となるように加水するのがさらに好ましい。 The hydration treatment as a (selective) reduction treatment of usnic acid will be described.
Usnic acid has high crystallinity and low solubility in aqueous solutions. For this reason, when the Sarcophagus lichen extract extracted using the extraction solvent defined in the present invention is stored in a solution having a high water content, usnic acid is hardly dissolved and crystals of usnic acid are precipitated. By removing the precipitated usnic acid by a conventional method, it becomes possible to (selectively) reduce the usnic acid from the Lichenaceae lichen extract.
In the first embodiment of the present invention, the amount of water added to the extract of Lichenaceae lichen can be appropriately set. In the second embodiment of the present invention, the amount of water added to the Saruogase lichen extract can be appropriately set within a range in which difractic acid is not lost due to precipitation or the like after addition. Specifically, it is preferable to add water so that the alcohol content after addition is 25% by volume or more, more preferably 30% by volume or more, and more preferably 45% by volume or less. It is preferable to add water, and it is more preferable to add water so as to be 40 volume% or less. Alternatively, it is preferable to add water so that the alcohol content after addition is 25 to 45% by volume, and it is more preferable to add 30% to 40% by volume.
In the third embodiment of the present invention, it is possible to appropriately set the amount of water added to the Sarcophagus lichen extract, and it is preferable to add water so that the alcohol content after addition is 25% by volume or more, and 30 volumes. It is more preferable to add water so as to be not less than 50%, preferably not more than 50% by volume, and more preferably not more than 45% by volume. Alternatively, it is preferable to add water so that the alcohol content after addition is 25 to 50% by volume, and it is more preferable to add 30% to 45% by volume.
　ウスニン酸の（選択的）低減処理としての活性炭処理について説明する。
　分子サイズ及び化合物の物性（極性）の違いから、サルオガセ属地衣類抽出物に含まれる成分のうち、ウスニン酸は活性炭に特に吸着されやすい。例えば、ジフラクタ酸とウスニン酸とでは、分子サイズ及び化合物の物性（極性）が異なる。これらの違いから、ウスニン酸の方がジフラクタ酸よりも活性炭に吸着されやすい。したがって、本発明の方法で得られるサルオガセ属地衣類抽出物に活性炭処理を施すことで、抽出物中のウスニン酸を（選択的に）除去することができる。
　活性炭の使用量は適宜設定することができ、サルオガセ属地衣類抽出物の固形分に対して、１重量％以上が好ましく、１０重量％以上がより好ましく、５０重量％以上がさらに好ましく、２００重量％以下が好ましく、１５０重量％以下がさらに好ましい。あるいは、１～２００重量％が好ましく、１０～２００重量％がより好ましく、５０～１５０重量％がさらに好ましい。また、処理時間も適宜設定することができ、１０分以上が好ましく、３０分以上がより好ましく、２４時間以下が好ましく、１時間以下がより好ましい。あるいは、１０分～２４時間が好ましく、３０分～１時間がより好ましい。活性炭処理を行う際、サルオガセ属地衣類抽出物と活性炭とを混合して撹拌してもよい。本発明において、活性炭としては通常のものでよく、市販のものを用いることができる。 The activated carbon treatment as a (selective) reduction treatment of usnic acid will be described.
Of the components contained in the extract of Ligocephalus lichen due to the difference in molecular size and physical properties (polarity) of the compound, usnic acid is particularly easily adsorbed on activated carbon. For example, difractic acid and usnic acid differ in molecular size and compound physical properties (polarity). Because of these differences, usnic acid is more easily adsorbed on activated carbon than difractic acid. Therefore, by applying the activated carbon treatment to the extract of the genus Sarhogase obtained by the method of the present invention, usnic acid in the extract can be (selectively) removed.
The amount of the activated carbon can be appropriately set, and is preferably 1% by weight or more, more preferably 10% by weight or more, still more preferably 50% by weight or more, and 200% by weight with respect to the solid content of the extract of Lichenaceae. The following is preferable, and 150% by weight or less is more preferable. Alternatively, it is preferably 1 to 200% by weight, more preferably 10 to 200% by weight, and further preferably 50 to 150% by weight. The treatment time can also be set as appropriate, preferably 10 minutes or longer, more preferably 30 minutes or longer, preferably 24 hours or shorter, more preferably 1 hour or shorter. Alternatively, 10 minutes to 24 hours are preferable, and 30 minutes to 1 hour are more preferable. When performing the activated carbon treatment, the Saruogase lichen extract and activated carbon may be mixed and stirred. In the present invention, the activated carbon may be a normal one, and a commercially available one can be used.
　ウスニン酸の（選択的）低減処理としてのヘキサン洗浄処理について説明する。
　サルオガセ属地衣類抽出物に含まれる成分のうち、ウスニン酸は特にヘキサンに溶解しやすい。また、ジフラクタ酸はヘキサンに溶解しづらいのに対し、ウスニン酸はヘキサンに溶解しやすい。そのため、サルオガセ属地衣類抽出物に対してヘキサン洗浄処理し、ヘキサン層を分離することで、サルオガセ属地衣類抽出物中のウスニン酸を（選択的に）除去することができる。
　ヘキサン洗浄処理の方法は適宜選択することができ、サルオガセ属地衣類抽出物及びヘキサン、並びに必要によりエタノールなどのアルコール溶液と混合して、２層に分離し、サルオガセ属地衣類抽出物を含む層を常法により回収すればよい。また、ヘキサン洗浄処理を繰返し行ってもよい。 A hexane washing process as a (selective) reduction process of usnic acid will be described.
Of the components contained in the Salhogase lichen extract, usnic acid is particularly easily dissolved in hexane. In addition, difractic acid is difficult to dissolve in hexane, whereas usnic acid is easily dissolved in hexane. For this reason, the usanic acid lichen extract is subjected to hexane washing treatment and the hexane layer is separated to remove (selectively) usnic acid in the sorghum lichen extract.
The method of washing with hexane can be selected as appropriate. The hexane washing lichen extract and hexane, and optionally mixed with an alcohol solution such as ethanol, are separated into two layers, and the layer containing the lichen extract is usually added. It may be recovered by the law. Further, the hexane cleaning process may be repeated.
　ウスニン酸の（選択的）低減処理としての加熱処理について説明する。
　サルオガセ属地衣類抽出物に対して加熱処理を行うことで、サルオガセ属地衣類抽出物中の他の成分とウスニン酸とが反応し、ウスニン酸の分解等が起こり、ウスニン酸の含有量が低下すると考えられる。
　サルオガセ属地衣類抽出物の加熱温度としては５０℃以上が好ましく、６０℃以上がさらに好ましく、１００℃以下が好ましく、９０℃以下がより好ましく、８０℃以下がさらに好ましい。あるいは、５０℃～１００℃が好ましく、５０℃～９０℃がより好ましく、６０℃～８０℃がさらに好ましい。また、サルオガセ属地衣類抽出物の加熱時間は加熱温度により適宜設定でき、１時間以上が好ましく、６時間以上がより好ましく、７日以下が好ましく、３日以下がより好ましい。あるいは、１時間～７日間が好ましく、６時間～３日間がより好ましい。 The heat treatment as a (selective) reduction treatment of usnic acid will be described.
It is thought that heat treatment is performed on the extract of Lichenaceae liquor, the other components in the lichen extract of Lilium extract react with usnic acid, decomposition of usnic acid occurs, and the content of usnic acid decreases. It is done.
The heating temperature of the extract of Lichenaceae is preferably 50 ° C. or higher, more preferably 60 ° C. or higher, preferably 100 ° C. or lower, more preferably 90 ° C. or lower, and further preferably 80 ° C. or lower. Alternatively, 50 ° C to 100 ° C is preferable, 50 ° C to 90 ° C is more preferable, and 60 ° C to 80 ° C is further preferable. Moreover, the heating time of the Saruogase lichen extract can be appropriately set depending on the heating temperature, preferably 1 hour or more, more preferably 6 hours or more, preferably 7 days or less, and more preferably 3 days or less. Alternatively, it is preferably 1 hour to 7 days, more preferably 6 hours to 3 days.
　本発明の第１の実施態様において、抽出溶媒としてアルコール含有率が８０体積％以下のアルコール水溶液を用いて得られるサルオガセ属地衣類抽出物に対して、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理及び加熱処理からなる群より選ばれる１種以上の処理によりウスニン酸のさらなる低減処理を行うことが好ましく、より好ましいウスニン酸の低減処理方法としては、下記のものが挙げられる。
（１）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理及び低温保存処理を行う。
（２）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理を行う。
（３）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理、加水処理及び低温保存処理をこの順で行う。
（４）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理及び低温保存処理をこの順で行う。
（５）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理、低温保存処理及び活性炭処理をこの順で行う。
（６）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理、低温保存処理及びヘキサン洗浄処理をこの順で行う。
（７）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理及び活性炭処理をこの順で行う。
（８）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理及びヘキサン洗浄処理をこの順で行う。
（９）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理、低温保存処理及び加熱処理をこの順で行う。 In the first embodiment of the present invention, a low-temperature storage treatment, a water addition treatment, an activated carbon treatment, and a hexane washing is performed on a Salgogase lichen extract obtained using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent. It is preferable to further reduce usnic acid by one or more treatments selected from the group consisting of treatment and heat treatment, and more preferable methods for reducing usnic acid include the following.
(1) The extracted Saruogase lichen extract is further subjected to a hydration treatment and a low-temperature storage treatment.
(2) A low-temperature storage treatment is further performed on the extracted Sarhogase lichen extract.
(3) A low temperature preservation process, a hydration process, and a low temperature preservation process are further performed in this order with respect to the extracted Sarhogase lichen extract.
(4) Further, a hydrotreating process and a low-temperature storage process are performed in this order on the extracted Lichenaceae lichen extract.
(5) Further, a hydrous treatment, a low-temperature storage treatment, and an activated carbon treatment are performed in this order on the extracted Saruogase lichen extract.
(6) Further, a hydrous treatment, a low-temperature storage treatment, and a hexane washing treatment are performed in this order on the extracted Sarhogase lichen extract.
(7) The low-temperature storage treatment and the activated carbon treatment are further performed in this order on the extracted Lichenaceae lichen extract.
(8) The low-temperature storage treatment and the hexane washing treatment are further performed in this order on the extracted Sarhogase lichen extract.
(9) The extracted Saruogase lichen extract is further subjected to a water treatment, a low-temperature storage treatment, and a heat treatment in this order.
　本発明の第１の実施態様の製造方法により得られるサルオガセ属地衣類抽出物において、ウスニン酸の絶対量は適宜設定することができ、ウスニン酸含有量は１５０ｐｐｍ以下であることが好ましく、１２０ｐｐｍ以下であることがさらに好ましい。 In the Sarcophagus lichen extract obtained by the production method of the first embodiment of the present invention, the absolute amount of usnic acid can be appropriately set, and the usnic acid content is preferably 150 ppm or less, preferably 120 ppm or less. More preferably it is.
　本発明の第２の実施態様において、抽出溶媒としてアルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて得られるサルオガセ属地衣類抽出物に対して、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理及び加熱処理からなる群より選ばれる１種以上の処理によりウスニン酸の選択的低減処理を行うことが好ましく、より好ましいウスニン酸の選択的低減処理方法としては、下記のものが挙げられる。
（１）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理及び低温保存処理を行う。
（２）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理を行う。
（３）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理、加水処理及び低温保存処理をこの順で行う。
（４）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理及び低温保存処理をこの順で行う。
（５）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理、低温保存処理及び活性炭処理をこの順で行う。
（６）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理、低温保存処理及びヘキサン洗浄処理をこの順で行う。
（７）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理及び活性炭処理をこの順で行う。
（８）抽出したサルオガセ属地衣類抽出物に対してさらに低温保存処理及びヘキサン洗浄処理をこの順で行う。
（９）抽出したサルオガセ属地衣類抽出物に対してさらに加水処理、低温保存処理及び加熱処理をこの順で行う。 In the second embodiment of the present invention, a low-temperature storage treatment, a water addition treatment, activated carbon is applied to a Salugatace lichen extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume as an extraction solvent. It is preferable to carry out selective reduction treatment of usnic acid by one or more treatments selected from the group consisting of treatment, hexane washing treatment and heat treatment, and more preferable methods for selective reduction of usnic acid are as follows. Can be mentioned.
(1) The extracted Saruogase lichen extract is further subjected to a hydration treatment and a low-temperature storage treatment.
(2) A low-temperature storage treatment is further performed on the extracted Sarhogase lichen extract.
(3) A low temperature preservation process, a hydration process, and a low temperature preservation process are further performed in this order with respect to the extracted Sarhogase lichen extract.
(4) Further, a hydrotreating process and a low-temperature storage process are performed in this order on the extracted Lichenaceae lichen extract.
(5) Further, a hydrous treatment, a low-temperature storage treatment, and an activated carbon treatment are performed in this order on the extracted Saruogase lichen extract.
(6) Further, a hydrous treatment, a low-temperature storage treatment, and a hexane washing treatment are performed in this order on the extracted Sarhogase lichen extract.
(7) The low-temperature storage treatment and the activated carbon treatment are further performed in this order on the extracted Lichenaceae lichen extract.
(8) The low-temperature storage treatment and the hexane washing treatment are further performed in this order on the extracted Sarhogase lichen extract.
(9) The extracted Saruogase lichen extract is further subjected to a water treatment, a low-temperature storage treatment, and a heat treatment in this order.
　本発明の第２の実施態様の製造方法により得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤において、ウスニン酸に対するジフラクタ酸の含有比率は適宜設定することができ、得られたサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が３以上であるのが好ましく、５以上がより好ましい。また、ウスニン酸の絶対量は適宜設定することができ、ウスニン酸含有量は１５０ｐｐｍ以下であることが好ましく、１２０ｐｐｍ以下であることがさらに好ましい。 In the Sarcophagus lichen extract and endothelin action inhibitor obtained by the production method of the second embodiment of the present invention, the content ratio of difractic acid relative to usnic acid can be set as appropriate, and the obtained Sarcoses lichen extract obtained. Alternatively, when the usnic acid content in the endothelin action inhibitor is set to 1 on a mass basis, the difractic acid content is preferably 3 or more, more preferably 5 or more. Moreover, the absolute amount of usnic acid can be set as appropriate, and the usnic acid content is preferably 150 ppm or less, and more preferably 120 ppm or less.
　本発明の第３の実施態様において、ウスニン酸の選択的低減処理により、サルオガセ属地衣類抽出物に含まれる、エンドセリン作用抑制剤、メラニン生成阻害剤、美白剤などの有効成分の含有量が低下しても、ウスニン酸の残存率が十分に低減されていればよい。例えば、本発明の第５の実施態様において、ウスニン酸の選択的低減処理によりサルオガセ属地衣類抽出物中のジフラクタ酸含有量が低下しても、ジフラクタ酸の残存率に対してウスニン酸の残存率が十分に低ければよい。 In the third embodiment of the present invention, the content of active ingredients such as endothelin action inhibitor, melanin production inhibitor, whitening agent, etc. contained in the extract of Lichenaceae by selective reduction of usnic acid is reduced. However, it is sufficient that the residual ratio of usnic acid is sufficiently reduced. For example, in the fifth embodiment of the present invention, even if the difractic acid content in the extract of Lichenaceae is reduced by the selective reduction treatment of usnic acid, the residual ratio of usnic acid relative to the residual ratio of difractic acid Should be low enough.
　本発明の第３の実施態様において、前記アルコール水溶液を用いた抽出、前記加水処理、及び前記低温保存処理をこの順で行うことが好ましい。また、前記アルコール水溶液を用いた抽出、前記低温保存処理、前記加水処理、及び前記低温保存処理をこの順で行うことも好ましい。 In the third embodiment of the present invention, it is preferable to perform the extraction using the aqueous alcohol solution, the hydration treatment, and the low-temperature storage treatment in this order. It is also preferable to perform the extraction using the aqueous alcohol solution, the low-temperature storage treatment, the hydration treatment, and the low-temperature storage treatment in this order.
　本発明の第３の実施態様の製造方法により得られるサルオガセ属地衣類抽出物において、ウスニン酸に対するジフラクタ酸の含有比率は適宜設定することができ、得られたサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が２以上であるのが好ましく、１０以上がより好ましい。また、ウスニン酸の絶対量は適宜設定することができ、ウスニン酸含有量は３００ｐｐｍ以下であることが好ましく、２００ｐｐｍ以下であることがさらに好ましい。 In the extract of Lichenaceae lichen obtained by the production method of the third embodiment of the present invention, the content ratio of difractic acid to usnic acid can be appropriately set, and the extract of Lizoaceae lichen or endothelin action inhibitor obtained When the content of usnic acid is 1 on the mass basis, the content of difractic acid is preferably 2 or more, and more preferably 10 or more. The absolute amount of usnic acid can be set as appropriate, and the usnic acid content is preferably 300 ppm or less, more preferably 200 ppm or less.
　本発明、特に本発明の第１及び第２の実施態様において、前記アルコール水溶液を用いてサルオガセ属地衣類の抽出を行う前に水系溶媒を用いてサルオガセ属地衣類を処理し、前記水系溶媒を除去して得られるサルオガセ属地衣類の残渣に対して前記アルコール水溶液を用いた抽出を行ってもよい。水系溶媒を用いてサルオガセ属地衣類を処理することによりサルオガセ属地衣類に含まれる水溶性成分を除去することができ、不純物が少ないサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤、又は不純物が少なくジフラクタ酸濃度が特に高いサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤を製造することができる。 In the first and second embodiments of the present invention, in particular, in the first and second embodiments of the present invention, the aqueous solution is used to treat the sorghum lichen before the extraction of the sorghum lichen using the aqueous alcohol solution, and the aqueous solvent is removed. Extraction using the aqueous alcohol solution may be performed on the residue of the Lichenaceae lichen obtained in this way. A water-soluble component contained in the sorghum lichen can be removed by treating the sorghum lichen with an aqueous solvent, and the extract of the sorghum lichen extract or endothelin action inhibitor with less impurities, or difractic acid concentration with less impurities Can be produced, particularly a Sargogase lichen extract or endothelin action inhibitor.
　本明細書において「水系溶媒」としては、サルオガセ属地衣類に含まれる水溶性成分を除去するだけの含水率を含む溶媒を適宜選択することができ、水、アルコール水溶液（例えば、メタノール水溶液、エタノール水溶液、プロパノール水溶液、イソプロパノール水溶液）等が挙げられる。このうち、水、又は、アルコール含有率が１０体積％以下、好ましくは５体積％以下、より好ましくは１体積％以下、のアルコール水溶液が好ましい。 In the present specification, as the “aqueous solvent”, a solvent containing a water content sufficient to remove water-soluble components contained in the genus Lisaceae can be appropriately selected. Water, an alcohol aqueous solution (for example, an aqueous methanol solution, an aqueous ethanol solution) Propanol aqueous solution, isopropanol aqueous solution) and the like. Among these, water or an alcohol aqueous solution having an alcohol content of 10% by volume or less, preferably 5% by volume or less, more preferably 1% by volume or less is preferable.
　前記水系溶媒を用いたサルオガセ属地衣類の処理方法については任意の方法を選択することができ、水系溶媒にサルオガセ属地衣類を浸漬させる方法、水系溶媒でサルオガセ属地衣類を洗浄する方法、などが挙げられる。
　水系溶媒にサルオガセ属地衣類を浸漬させる場合は、サルオガセ属地衣類に含まれる水溶性成分を除去する効率を上げるため、処理の際に攪拌を行ったり、溶媒中でホモジナイズ処理を行ってもよい。 Arbitrary methods can be selected as a method for treating S. aureus lichen using the above-mentioned aqueous solvent, and a method of immersing S. aureus lichen in an aqueous solvent, a method of washing a sarcoma lichen with an aqueous solvent, etc. .
In the case of immersing the sorghum lichen in an aqueous solvent, in order to increase the efficiency of removing water-soluble components contained in the sorghum lichen, stirring may be performed during the treatment or homogenization treatment may be performed in the solvent.
　水系溶媒を用いたサルオガセ属地衣類の処理条件については、使用する溶媒によって適宜設定できる。例えば、処理温度は、０℃以上が好ましく、５℃以上がより好ましく、５０℃以下が好ましく、３０℃以下がより好ましい。あるいは、０～５０℃が好ましく、０～３０℃がより好ましい。処理時間は、５時間以上が好ましく、１日間以上がより好ましく、１０日間以下が好ましく、７日間以下がより好ましい。あるいは、５時間～１０日間が好ましく、１～７日間がより好ましい。 The treatment conditions for Sargogase lichen using an aqueous solvent can be appropriately set depending on the solvent used. For example, the treatment temperature is preferably 0 ° C. or higher, more preferably 5 ° C. or higher, preferably 50 ° C. or lower, and more preferably 30 ° C. or lower. Alternatively, 0 to 50 ° C. is preferable, and 0 to 30 ° C. is more preferable. The treatment time is preferably 5 hours or longer, more preferably 1 day or longer, preferably 10 days or shorter, and more preferably 7 days or shorter. Alternatively, it is preferably 5 hours to 10 days, and more preferably 1 to 7 days.
　本発明、特に本発明の第１及び第２の実施態様において、前記アルコール水溶液を用いてサルオガセ属地衣類の抽出を行う前にヘキサン等の疎水性有機溶媒を用いてサルオガセ属地衣類を処理し、ウスニン酸等の疎水性物質を除去して得られるサルオガセ属地衣類の残渣に対して前記アルコール水溶液を用いた抽出を行っても良い。疎水性有機溶媒を用いてサルオガセ属地衣類を処理することによりサルオガセ属地衣類に含まれる疎水性成分を除去することができ、不純物が少ないサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤、又は不純物が少なくジフラクタ酸濃度が特に高いサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤を製造することができる。 In the first and second embodiments of the present invention, in particular in the first and second embodiments of the present invention, before the extraction of S. agarus lichen using the aqueous alcohol solution, the S. aeruginosa lichen is treated with a hydrophobic organic solvent such as hexane. You may perform extraction using the said aqueous alcohol solution with respect to the residue of Salgogase lichen obtained by removing hydrophobic substances, such as an acid. Hydrophobic components contained in Sorghum lichen can be removed by treating Sorghum lichen using a hydrophobic organic solvent, and the extract of Lactobacillus lichen extract or endothelin action inhibitor with little impurities, or difractor with few impurities It is possible to produce an extract of Saruogase lichen or an endothelin action inhibitor having a particularly high acid concentration.
　本明細書において「疎水性有機溶媒」としては、サルオガセ属地衣類に含まれる疎水性成分を除去できる溶媒を適宜選択することができ、n-ペンタン、シクロペンタン、n-ヘキサン、シクロヘキサン、n-ヘプタン、シクロヘプタン、オクタン等の直鎖又は環状の常温、常圧で液体のアルカンが挙げられ、このうちn-ヘキサンが好ましい。 In the present specification, as the “hydrophobic organic solvent”, a solvent capable of removing the hydrophobic component contained in the garlic lichen can be appropriately selected, and n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane. , Cycloheptane, octane, and other linear alkanes that are liquid at room temperature and pressure, and n-hexane is preferred.
　前記疎水性有機溶媒を用いたサルオガセ属地衣類の処理方法については適宜選択することができ、疎水性有機溶媒にサルオガセ属地衣類を浸漬させる方法、疎水性有機溶媒でサルオガセ属地衣類を洗浄する方法、などが挙げられる。
　疎水性有機溶媒にサルオガセ属地衣類を浸漬させる場合は、サルオガセ属地衣類に含まれる疎水性物質を除去する効率を上げるため、処理の際に攪拌を行ったり、溶媒中でホモジナイズ処理を行ってもよい。 About the processing method of the Sarcophagus lichen using the said hydrophobic organic solvent, it can choose suitably, The method of immersing the Sarcophagus lichen in the hydrophobic organic solvent, The method of wash | cleaning Sargoses lichen with a hydrophobic organic solvent, etc. Is mentioned.
In the case of immersing the sorghum lichen in a hydrophobic organic solvent, in order to increase the efficiency of removing the hydrophobic substance contained in the sorghum lichen, stirring may be performed during the treatment or a homogenization treatment may be performed in the solvent. .
　疎水性有機溶媒を用いたサルオガセ属地衣類の処理条件については、使用する溶媒によって適宜設定できる。例えば、処理温度は、０℃以上が好ましく、５℃以上がより好ましく、５０℃以下が好ましく、３０℃以下がより好ましい。あるいは、０～５０℃が好ましく、０～３０℃がより好ましい。処理時間は、５時間以上が好ましく、１日間以上がより好ましく、１０日間以下が好ましく、７日間以下がより好ましい。あるいは、５時間～１０日間が好ましく、１～７日間がより好ましい。 The processing conditions for the Sarcophagus lichen using a hydrophobic organic solvent can be appropriately set depending on the solvent used. For example, the treatment temperature is preferably 0 ° C. or higher, more preferably 5 ° C. or higher, preferably 50 ° C. or lower, and more preferably 30 ° C. or lower. Alternatively, 0 to 50 ° C. is preferable, and 0 to 30 ° C. is more preferable. The treatment time is preferably 5 hours or longer, more preferably 1 day or longer, preferably 10 days or shorter, and more preferably 7 days or shorter. Alternatively, it is preferably 5 hours to 10 days, and more preferably 1 to 7 days.
　本発明の第１及び第２の実施態様について、後述の実施例で示すように、前記抽出工程により、肝毒性物質であるウスニン酸の含有量が低いナガサルオガセ抽出物を得ることができる。したがって、ナガサルオガセ抽出物はメラニン生成阻害作用や後述の実施例で示すエンドセリン作用抑制作用を有するため、本発明の第１の実施態様の製造方法で得られるウスニン酸低含有サルオガセ属地衣類抽出物は安全性が高く、エンドセリン抑制剤、メラニン生成阻害剤など、美白効果を奏する皮膚外用剤に好適に用いることができる。 As for the first and second embodiments of the present invention, as shown in the examples described later, Nagasagaogase extract having a low content of usnic acid, which is a hepatotoxic substance, can be obtained by the extraction step. Therefore, Nagasagaogase extract has a melanin production inhibitory action and an endothelin action inhibitory action shown in the examples described later. Therefore, the Ususic acid low-content Salogase lichen extract obtained by the production method of the first embodiment of the present invention is It is highly safe and can be suitably used for an external preparation for skin that exhibits a whitening effect, such as an endothelin inhibitor and a melanin production inhibitor.
　本発明の第２の実施態様について、後述の実施例で示すように、前記抽出工程により、ジフラクタ酸の含有量が高くウスニン酸の含有量が低いサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤を得ることができる。したがって、ジフラクタ酸はメラノサイトに対するエンドセリンの作用を抑制することでメラニン生成を抑制しうるため、本発明の第２の実施態様の製造方法で得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤は、優れたエンドセリン作用抑制効果及び美白効果を奏する。 About the 2nd embodiment of this invention, as shown in the below-mentioned Example, the extraction process WHEREIN: The Salomasae lichen extract and the endothelin action inhibitor which are high in the content of difractic acid and low in the content of usnic acid are obtained. be able to. Therefore, since difractic acid can suppress the production of melanin by suppressing the action of endothelin on melanocytes, the Saruogase lichen extract and endothelin action inhibitor obtained by the production method of the second embodiment of the present invention are excellent. It has an endothelin action inhibitory effect and a whitening effect.
　本発明の第３の実施態様について、後述の実施例で示すように、本発明の第３の実施態様により、ウスニン酸の含有量を十分に低下させたサルオガセ属地衣類抽出物を得ることができる。また、本発明の第５の実施態様によれば、ジフラクタ酸の含有量が高くウスニン酸の含有量が低いサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤を得ることができる。
　サルオガセ属地衣類抽出物は、メラニン生成阻害作用や後述の実施例で示すエンドセリン作用抑制効果を有する。また、ジフラクタ酸もエンドセリン作用抑制効果を有する。したがって、本発明の第３の実施態様の製造方法により得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤はメラノサイトに対するエンドセリンの作用を抑制することでメラニン生成を抑制しうるため、エンドセリン抑制剤、メラニン生成阻害剤など、美白効果を奏する皮膚外用剤に用いることができる。 About the 3rd embodiment of the present invention, as shown in the below-mentioned example, according to the 3rd embodiment of the present invention, the extract of the genus Sarugagase liquor which fully reduced the content of usnic acid can be obtained. . Further, according to the fifth embodiment of the present invention, it is possible to obtain an extract of Salgocase lichen and endothelin action inhibitor having a high difractic acid content and a low usnic acid content.
The Saruogase genus lichen extract has a melanin production inhibitory effect and an endothelin action inhibitory effect shown in Examples described later. Difractic acid also has an endothelin action inhibitory effect. Therefore, since the Sarcophagus lichen extract and endothelin action inhibitor obtained by the production method of the third embodiment of the present invention can suppress the production of melanin by inhibiting the action of endothelin on melanocytes, the endothelin inhibitor, melanin It can be used for an external preparation for skin that exhibits a whitening effect, such as a production inhibitor.
　本明細書において「美白（作用）」とは、メラニン色素生成や色素沈着を抑え、余分なメラニンのない本来の透明な肌色に戻すこと、または皮膚の黒化若しくはシミ・ソバカス等の色素沈着を防止、抑制することを意味する。
　また、本発明の第２及び第３の実施態様の製造方法により得られるサルオガセ属地衣類抽出物及びエンドセリン作用抑制剤は肝毒性物質であるウスニン酸の含有量が少ないため、前記サルオガセ属地衣類抽出物及びエンドセリン作用抑制剤は安全性が高く美白剤などの皮膚外用剤として好適に用いることができる。 In this specification, “whitening (action)” refers to suppressing melanin pigment formation and pigmentation, returning to an original transparent skin color without excess melanin, or pigmentation such as skin blackening or stain / sobacus. It means to prevent and suppress.
In addition, the sarcoma lichen extract obtained by the production method of the second and third embodiments of the present invention and the endothelin action inhibitor have a low content of usnic acid, which is a hepatotoxic substance. And endothelin action inhibitor has high safety and can be suitably used as a skin external preparation such as a whitening agent.
　本発明により得られるサルオガセ属地衣類抽出物は、そのままエンドセリン作用抑制剤又は美白剤として用いてもよい。また、本発明により得られるサルオガセ属地衣類抽出物と、例えば酸化チタン、炭酸カルシウム、蒸留水、乳糖、デンプン等の適当な液体または固体の賦形剤または増量剤とを混合して用いてもよい。さらには、その他のエンドセリン作用抑制剤、その他の美白剤、保湿剤、酸化防止剤、紫外線吸収剤、界面活性剤、増粘剤、色材種、油性物質、高分子化合物、防腐剤、粉体、色素、香料、乳化安定剤、pH調整剤等、通常の美白剤に用いられる成分と混合して用いてもよい。
　なお、本発明の第４の実施態様により得られるサルオガセ属地衣類抽出物をエンドセリン作用抑制剤又は美白剤に用いる場合、ジフラクタ酸を有効成分とせず、他の成分を有効成分としてもよい。また、本発明の第５の実施態様により得られるサルオガセ属地衣類抽出物をエンドセリン作用抑制剤又は美白剤に用いる場合、ジフラクタ酸のみを有効成分としてもよいし、ジフラクタ酸と他の成分と組み合わせて有効成分としてもよい。 The Saruogase lichen extract obtained by the present invention may be used as an endothelin inhibitor or a whitening agent as it is. In addition, the Lichenaceae lichen extract obtained by the present invention may be used by mixing with a suitable liquid or solid excipient or extender such as titanium oxide, calcium carbonate, distilled water, lactose, starch and the like. . Furthermore, other endothelin action inhibitors, other whitening agents, moisturizers, antioxidants, UV absorbers, surfactants, thickeners, colorants, oily substances, polymer compounds, preservatives, powders , Pigments, fragrances, emulsion stabilizers, pH adjusters, and the like, may be used in admixture with components used in ordinary whitening agents.
In addition, when using the Saruogase genus lichen extract obtained by the 4th embodiment of this invention for an endothelin action inhibitor or a whitening agent, it is good also considering difractic acid as an active ingredient and making another ingredient into an active ingredient. In addition, when using the Sarhogase lichen extract obtained by the fifth embodiment of the present invention as an endothelin action inhibitor or whitening agent, only difractic acid may be used as an active ingredient, or in combination with difractic acid and other ingredients. It may be an active ingredient.
　本発明の方法により得られるエンドセリン作用抑制剤及び本発明の美白剤は、皮膚化粧料、外用医薬品、外用医薬部外品等として皮膚等に適用することができ、その剤型も水溶液系、可溶化系、乳化系、粉末系、ゲル系、軟膏系、クリーム、水－油２層系、水－油－粉末３層系など、幅広い形態をとり得る。例えば、洗顔料、化粧水、乳液、クリーム、ローション、ジェル、エッセンス（美容液）、パック、マスク、ファンデーション、軟膏、シート状製品等の形態が挙げられる。 The endothelin action inhibitor obtained by the method of the present invention and the whitening agent of the present invention can be applied to the skin and the like as skin cosmetics, external medicines, quasi-drugs, etc. It can take a wide variety of forms such as a solubilization system, an emulsification system, a powder system, a gel system, an ointment system, a cream, a water-oil two-layer system, and a water-oil-powder three-layer system. Examples thereof include face wash, lotion, milky lotion, cream, lotion, gel, essence (beauty serum), pack, mask, foundation, ointment, sheet-like product and the like.
　本発明の方法により得られるエンドセリン作用抑制剤及び本発明の美白剤におけるサルオガセ属地衣類抽出物の含有量は適宜設定することができる。例えば、サルオガセ属地衣類抽出物中のジフラクタ酸などの有効成分濃度に依存して適宜設定することができる。
　また、本発明の方法により得られるエンドセリン作用抑制剤及び本発明の美白剤の使用量は適宜設定することができ、有効成分の含有量により異なるが、例えばクリーム状、軟膏状の場合、皮膚面１ｃｍ2当たり０．１μｇ以上が好ましく、５μｇ以下が好ましい。あるいは、皮膚面１ｃｍ2当たり０．１～５μｇが好ましい。液状製剤の場合、皮膚面１ｃｍ2当たり０．１μｇ以上が好ましく、１０μｇ以下が好ましい。あるいは、皮膚面１ｃｍ2当たり０．１～１０μｇが好ましい。 The content of the Sargogus genus lichen extract in the endothelin action inhibitor obtained by the method of the present invention and the whitening agent of the present invention can be appropriately set. For example, it can be set as appropriate depending on the concentration of an active ingredient such as difractic acid in the extract of Lichenaceae lichen.
In addition, the amount of endothelin action inhibitor obtained by the method of the present invention and the amount of whitening agent of the present invention can be appropriately set and varies depending on the content of the active ingredient. For example, in the case of cream or ointment, the skin surface 0.1 μg or more per 1 cm 2 is preferable, and 5 μg or less is preferable. Alternatively, 0.1 to 5 μg per 1 cm 2 of the skin surface is preferable. In the case of a liquid preparation, 0.1 μg or more per 1 cm 2 of skin surface is preferable, and 10 μg or less is preferable. Alternatively, 0.1 to 10 μg per 1 cm 2 of the skin surface is preferable.
　上述した実施形態に関し、本発明はさらに以下の抽出物の製造方法、エンドセリン作用抑制剤、美白剤、方法及び使用を開示する。 Regarding the above-described embodiment, the present invention further discloses the following extract production method, endothelin action inhibitor, whitening agent, method and use.
＜１＞サルオガセ属地衣類を、アルコール含有率が８０体積％以下、好ましくは７０体積％以下、より好ましくは６０体積％以下、のアルコール水溶液を用いて抽出する、ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。
＜２＞サルオガセ属地衣類を、アルコール含有率が３０体積％以上、好ましくは４０体積％以上、かつ８０体積％以下、好ましくは７０体積％以下、より好ましくは６０体積％以下、のアルコール水溶液を用いて抽出する、ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。 <1> A Salmonella lichen extract containing a low amount of usnic acid, which is extracted using an aqueous alcohol solution having an alcohol content of 80% by volume or less, preferably 70% by volume or less, more preferably 60% by volume or less. Manufacturing method.
<2> Using the alcoholic aqueous solution having an alcohol content of 30% by volume or more, preferably 40% by volume or more and 80% by volume or less, preferably 70% by volume or less, more preferably 60% by volume or less. A method for producing a lichenaceae lichen extract containing a high content of difractic acid and a low content of usnic acid.
＜３＞前記アルコール水溶液がエタノール水溶液又は1,3-ブタンジオール水溶液である、前記＜１＞又は＜２＞項記載の製造方法。
＜４＞前記サルオガセ属地衣類がナガサルオガセ、ヨコワサルオガセ、フジサルオガセ、ニセフジサルオガセ及びクシノハサルオガセからなる群より選ばれる少なくとも１種、好ましくはナガサルオガセ及びヨコワサルオガセからなる群より選ばれる少なくとも１種である、前記＜１＞～＜３＞のいずれか１項記載の製造方法。
＜５＞抽出したサルオガセ属地衣類抽出物に対して、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理及び加熱処理からなる群より選ばれる１種以上の処理によりウスニン酸の（選択的）低減処理を行う、前記＜１＞～＜４＞のいずれか１項記載の製造方法。
＜６＞抽出したサルオガセ属地衣類抽出物に対して加水処理及び低温保存処理を施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜７＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理を施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜８＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理、加水処理及び低温保存処理をこの順で施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜９＞抽出したサルオガセ属地衣類抽出物に対して加水処理及び低温保存処理をこの順で施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜１０＞抽出したサルオガセ属地衣類抽出物に対して加水処理、低温保存処理及び活性炭処理をこの順で施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜１１＞抽出したサルオガセ属地衣類抽出物に対して加水処理、低温保存処理及びヘキサン洗浄処理をこの順で施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜１２＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理及び活性炭処理をこの順で施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜１３＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理及びヘキサン洗浄処理をこの順で施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜１４＞抽出したサルオガセ属地衣類抽出物に対して加水処理、低温保存処理及び加熱処理をこの順で施して前記ウスニン酸の（選択的）低減処理を行う、前記＜５＞項記載の製造方法。
＜１５＞水系溶媒（好ましくは水、又はアルコール含有率が１０体積％以下（好ましくは５体積％以下、より好ましくは１体積％以下）のアルコール水溶液（好ましくは、メタノール水溶液、エタノール水溶液、プロパノール水溶液、又はイソプロパノール水溶液））を用いて前記サルオガセ属地衣類を処理し、処理したサルオガセ属地衣類に対して前記アルコール水溶液を用いた抽出を行う、前記＜１＞～＜１４＞のいずれか１項記載の製造方法。
＜１６＞疎水性有機溶媒（好ましくはn-ペンタン、シクロペンタン、n-ヘキサン、シクロヘキサン、n-ヘプタン、シクロヘプタン及びオクタンからなる群より選ばれる少なくとも１種の疎水性有機溶媒、より好ましくはn-ヘキサン）を用いて前記サルオガセ属地衣類を処理し、処理したサルオガセ属地衣類に対して前記アルコール水溶液を用いた抽出を行う、前記＜１＞～＜１５＞のいずれか１項記載の製造方法。
＜１７＞得られた抽出物中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が３以上、好ましくは５以上、である、前記＜２＞～＜１６＞のいずれか１項記載の製造方法。 <3> The method according to <1> or <2>, wherein the alcohol aqueous solution is an ethanol aqueous solution or a 1,3-butanediol aqueous solution.
<4> At least one selected from the group consisting of Nagasaga Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefu Gisal Ogase and Kushinohasaru Ogase, preferably at least one selected from the group consisting of Nagasaro Ogase and Yokosawa Ogase The production method according to any one of <1> to <3>, wherein
<5> (selective) reduction of usnic acid by one or more treatments selected from the group consisting of low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment, and heat treatment on the extracted Lichenaceae lichen extract The production method according to any one of <1> to <4>, wherein the treatment is performed.
<6> The method according to <5> above, wherein the extracted extract of Sorghum lichen is subjected to water treatment and low-temperature storage treatment to perform (selective) reduction of the usnic acid.
<7> The production method according to <5> above, wherein the extracted extract of the genus Sarhogase is subjected to a low-temperature storage treatment to perform a (selective) reduction treatment of the usnic acid.
<8> The production according to <5>, wherein the extracted extract of the genus Sarhogase is subjected to a low-temperature storage treatment, a hydration treatment, and a low-temperature storage treatment in this order to perform (selective) reduction of the usnic acid. Method.
<9> The production method according to <5>, wherein the extracted extract of the genus Sarhogase is subjected to water treatment and low-temperature storage treatment in this order to perform (selective) reduction of the usnic acid.
<10> The method according to <5>, wherein the extracted extract of the genus Sarhogase is subjected to water treatment, low-temperature storage treatment, and activated carbon treatment in this order to perform (selective) reduction of the usnic acid. .
<11> The production according to <5> above, wherein the extracted extract of Salgogase lichen is subjected to water treatment, low-temperature storage treatment, and hexane washing treatment in this order to perform (selective) reduction of the usnic acid. Method.
<12> The production method according to <5>, wherein the extracted extract of the genus Sarhogase is subjected to low-temperature storage treatment and activated carbon treatment in this order to perform (selective) reduction treatment of the usnic acid.
<13> The production method according to <5>, wherein the extracted sarcoma lichen extract is subjected to a low-temperature storage treatment and a hexane washing treatment in this order to perform (selective) reduction of the usnic acid.
<14> The method according to <5> above, wherein the extracted extract of the genus Sarhogase is subjected to water treatment, low-temperature storage treatment, and heat treatment in this order to perform (selective) reduction of the usnic acid. .
<15> Aqueous solvent (preferably water or an alcohol content of 10% by volume or less (preferably 5% by volume or less, more preferably 1% by volume or less)) (preferably methanol aqueous solution, ethanol aqueous solution, propanol aqueous solution) Or the aqueous solution of isopropanol)), and the extracted aqueous liquor is extracted with the aqueous alcohol solution, according to any one of the above items <1> to <14> Production method.
<16> a hydrophobic organic solvent (preferably at least one hydrophobic organic solvent selected from the group consisting of n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane, cycloheptane and octane, more preferably n The method according to any one of <1> to <15>, wherein the sarcoma lichen is treated with -hexane), and the treated sarcoma lichen is extracted with the aqueous alcohol solution.
<17> Any of the above <2> to <16>, wherein the content of usnic acid in the obtained extract is 1 on a mass basis, the difractic acid content is 3 or more, preferably 5 or more The manufacturing method of 1 item | term.
＜１８＞サルオガセ属地衣類を、アルコール含有率が７０体積％を超える、好ましくは８０体積％を超える、より好ましくは９０体積％以上、さらに好ましくは９５体積％以上、のアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う抽出物の製造方法であって、前記処理後のウスニン酸の残存率が５０％未満である、ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。
＜１９＞サルオガセ属地衣類を、アルコール含有率が７０体積％を超える、好ましくは８０体積％を超える、より好ましくは９０体積％以上、さらに好ましくは９５体積％以上、のアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う、ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。 <18> Salogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, preferably more than 80% by volume, more preferably 90% by volume or more, and even more preferably 95% by volume or more. A method for producing an extract, wherein the obtained extract is subjected to at least one treatment selected from the group consisting of a low-temperature storage treatment and a hydrolysis treatment, wherein the residual ratio of usnic acid after the treatment is less than 50% A method for producing an extract of Lichenaceae lichen extract containing low usnic acid.
<19> Salogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, preferably more than 80% by volume, more preferably 90% by volume or more, and even more preferably 95% by volume or more. A method for producing an extract of a Lichenaceae lichen extract containing a high content of difractic acid and a low content of usnic acid, wherein the obtained extract is subjected to at least one treatment selected from the group consisting of a low-temperature storage treatment and a hydrolysis treatment.
＜２０＞前記アルコール水溶液がエタノール水溶液である、前記＜１８＞又は＜１９＞項記載の製造方法。
＜２１＞前記サルオガセ属地衣類がナガサルオガセ、ヨコワサルオガセ、フジサルオガセ、ニセフジサルオガセ及びクシノハサルオガセからなる群より選ばれる少なくとも１種、好ましくはナガサルオガセ及びヨコワサルオガセからなる群より選ばれる少なくとも１種である、前記＜１８＞～＜２０＞のいずれか１項記載の製造方法。
＜２２＞前記アルコール水溶液を用いた抽出、前記加水処理、及び前記低温保存処理をこの順で行う、前記＜１８＞～＜２１＞のいずれか１項記載の製造方法。
＜２３＞前記アルコール水溶液を用いた抽出、前記低温保存処理、前記加水処理、及び前記低温保存処理をこの順で行う、前記＜１８＞～＜２２＞のいずれか１項記載の製造方法。
＜２４＞得られた抽出物中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が２以上、好ましくは１０以上、である、前記＜１９＞～＜２３＞のいずれか１項記載の製造方法。 <20> The method according to <18> or <19>, wherein the aqueous alcohol solution is an aqueous ethanol solution.
<21> At least one selected from the group consisting of Nagasaro-Ogase, Yokosawa-Ogase, Fujisaru-Ogase, Nisefu-jisaru-ogase and Kushinohasaru-ogase, preferably at least one selected from the group consisting of Nagasaro-ogase and Yokosawa-o-gase The production method according to any one of <18> to <20>, wherein
<22> The production method according to any one of <18> to <21>, wherein the extraction using the aqueous alcohol solution, the hydrolysis treatment, and the low-temperature storage treatment are performed in this order.
<23> The method according to any one of <18> to <22>, wherein the extraction using the aqueous alcohol solution, the low-temperature storage treatment, the hydration treatment, and the low-temperature storage treatment are performed in this order.
<24> Any one of the above <19> to <23>, wherein when the usnic acid content in the obtained extract is 1 on a mass basis, the difractic acid content is 2 or more, preferably 10 or more The manufacturing method of 1 item | term.
＜２５＞前記＜１＞～＜１６＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物を有効成分とする、エンドセリン作用抑制剤。
＜２６＞前記＜１＞～＜１６＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物を有効成分とする、美白剤。 <25> An endothelin action inhibitor comprising, as an active ingredient, an extract of the genus Sarhogase obtained by the production method according to any one of <1> to <16>.
<26> A whitening agent comprising, as an active ingredient, an extract of a Lichenaceae lichen obtained by the production method according to any one of <1> to <16>.
＜２７＞サルオガセ属地衣類を、アルコール含有率がアルコール含有率が３０体積％以上、好ましくは４０体積％以上、かつ８０体積％以下、好ましくは７０体積％以下、より好ましくは６０体積％以下、のアルコール水溶液を用いて抽出する、エンドセリン作用抑制剤の製造方法。 <27> Sarhogase lichen having an alcohol content of 30% by volume or more, preferably 40% by volume or more and 80% by volume or less, preferably 70% by volume or less, more preferably 60% by volume or less. The manufacturing method of an endothelin action inhibitor extracted using alcohol aqueous solution.
＜２８＞前記アルコール水溶液がエタノール水溶液又は1,3-ブタンジオール水溶液である、前記＜２７＞項記載の製造方法。
＜２９＞前記サルオガセ属地衣類がナガサルオガセ、ヨコワサルオガセ、フジサルオガセ、ニセフジサルオガセ及びクシノハサルオガセからなる群より選ばれる少なくとも１種、好ましくはナガサルオガセ及びヨコワサルオガセからなる群より選ばれる少なくとも１種である、前記＜２７＞又は＜２８＞項記載の製造方法。
＜３０＞抽出したサルオガセ属地衣類抽出物に対して、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理及び加熱処理からなる群より選ばれる１種以上の処理により該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜２７＞～＜２９＞のいずれか１項記載の製造方法。
＜３１＞抽出したサルオガセ属地衣類抽出物に対して加水処理及び低温保存処理を施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３２＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理を施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３３＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理、加水保存処理及び低温保存処理をこの順で施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３４＞抽出したサルオガセ属地衣類抽出物に対して加水処理及び低温保存処理をこの順で施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３５＞抽出したサルオガセ属地衣類抽出物に対して加水処理、低温保存処理及び活性炭処理をこの順で施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３６＞抽出したサルオガセ属地衣類抽出物に対して加水処理、低温保存処理及びヘキサン洗浄処理をこの順で施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３７＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理及び活性炭処理をこの順で施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３８＞抽出したサルオガセ属地衣類抽出物に対して低温保存処理及びヘキサン洗浄処理をこの順で施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜３９＞抽出したサルオガセ属地衣類抽出物に対して加水処理、低温保存処理及び加熱処理をこの順で施して該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、前記＜３０＞項記載の製造方法。
＜４０＞水系溶媒（好ましくは水、又はアルコール含有率が１０体積％以下（好ましくは５体積％以下、より好ましくは１体積％以下）のアルコール水溶液（好ましくは、メタノール水溶液、エタノール水溶液、プロパノール水溶液、又はイソプロパノール水溶液））を用いて前記サルオガセ属地衣類を処理し、処理したサルオガセ属地衣類に対して前記アルコール水溶液を用いた抽出を行う、前記＜２７＞～＜３９＞のいずれか１項記載の製造方法。
＜４１＞疎水性有機溶媒（好ましくはn-ペンタン、シクロペンタン、n-ヘキサン、シクロヘキサン、n-ヘプタン、シクロヘプタン及びオクタンからなる群より選ばれる少なくとも１種の疎水性有機溶媒、より好ましくはn-ヘキサン）を用いて前記サルオガセ属地衣類を処理し、処理したサルオガセ属地衣類に対して前記アルコール水溶液を用いた抽出を行う、前記＜２７＞～＜４０＞のいずれか１項記載の製造方法。
＜４２＞得られたエンドセリン作用抑制剤中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が３以上、好ましくは５以上、である、前記＜２７＞～＜４１＞のいずれか１項記載の製造方法。 <28> The production method according to <27>, wherein the alcohol aqueous solution is an ethanol aqueous solution or a 1,3-butanediol aqueous solution.
<29> At least one selected from the group consisting of Nagasaga Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefu Gisal Ogase, and Kushinohasaru Ogase, preferably at least one selected from the group consisting of Nagasaro Ogase and Yokosawa Ogase The method according to <27> or <28>, wherein
<30> Extracted from the genus Lichenaceae liquor, extracted from the genus Lichenaceae liquor by one or more treatments selected from the group consisting of low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment and heat treatment The production method according to any one of <27> to <29>, wherein the selective reduction treatment of usnic acid is performed.
<31> The method according to <30> above, wherein the extracted extract of Lichenaceae liquor is subjected to a hydrolysis treatment and a low-temperature storage treatment to selectively reduce the usnic acid contained in the lichen extract of Lilium spp. .
<32> The production method according to <30>, wherein the extracted extract of the genus Sarhogase is subjected to a low-temperature storage treatment to selectively reduce the usnic acid contained in the extract of the genus Sargogase.
<33> A low-temperature preservation treatment, a hydrolyzing preservation treatment, and a low-temperature preservation treatment are performed in this order on the extracted Saruogase lichen extract in this order to perform a selective reduction treatment of usnic acid contained in the Saruogase lichen extract, <30> The production method according to item.
<34> The description of <30> above, wherein the extracted extract of the genus Sarhogase is subjected to a hydrolysis treatment and a low-temperature storage treatment in this order to selectively reduce the usnic acid contained in the extract of the genus Sarogase genus. Manufacturing method.
<35> The extract of Salgogase lichen is subjected to water treatment, low-temperature storage treatment and activated carbon treatment in this order to perform selective reduction treatment of usnic acid contained in the Salgogase lichen extract, <30 > The manufacturing method of description.
<36> The extract of Salgogase lichen is subjected to water treatment, low-temperature storage treatment, and hexane washing treatment in this order to perform a selective reduction treatment of usnic acid contained in the Salgogase lichen extract. Item 30>.
<37> The item <30> above, wherein the extracted Sarhogase lichen extract is subjected to a low-temperature storage treatment and an activated carbon treatment in this order to selectively reduce the usnic acid contained in the Salgogase lichen extract. Manufacturing method.
<38> The item <30> above, wherein a low-temperature storage treatment and a hexane washing treatment are performed in this order on the extracted Sargases lichen extract in this order to perform a selective reduction treatment of usnic acid contained in the Salgoses lichen extract. The manufacturing method as described.
<39> The extract of Salgogase lichen is subjected to water treatment, low-temperature storage treatment, and heat treatment in this order to perform a selective reduction treatment of usnic acid contained in the Salgogase lichen extract, <30 > The manufacturing method of description.
<40> an aqueous alcohol solution (preferably water or an alcohol content of 10% by volume or less (preferably 5% by volume or less, more preferably 1% by volume or less)) (preferably methanol aqueous solution, ethanol aqueous solution, propanol aqueous solution) Or the aqueous solution of isopropanol)), and the extracted aqueous liquor is extracted with the aqueous alcohol solution, according to any one of the above items <27> to <39> Production method.
<41> a hydrophobic organic solvent (preferably at least one hydrophobic organic solvent selected from the group consisting of n-pentane, cyclopentane, n-hexane, cyclohexane, n-heptane, cycloheptane and octane, more preferably n The production method according to any one of <27> to <40>, wherein the sarcoma lichen is treated using (-hexane), and the treated sarcophagus lichen is extracted using the aqueous alcohol solution.
<42> When the content of usnic acid in the obtained endothelin inhibitor is set to 1 on a mass basis, the content of difractic acid is 3 or more, preferably 5 or more, <27> to <41> The manufacturing method of any one of Claims.
＜４３＞前記＜１＞～＜１７＞及び＜２７＞～＜４２＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物又はエンドセリン作用抑制剤を有効成分とする、美白剤。 <43> A whitening agent comprising, as an active ingredient, a Salugose lichen extract or endothelin action inhibitor obtained by the production method according to any one of <1> to <17> and <27> to <42>.
＜４４＞サルオガセ属地衣類を、アルコール含有率が７０体積％を超える、好ましくは８０体積％を超える、より好ましくは９０体積％以上、さらに好ましくは９５体積％以上、のアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う、エンドセリン作用抑制剤の製造方法。 <44> Salogase lichen is extracted using an alcohol aqueous solution having an alcohol content of more than 70% by volume, preferably more than 80% by volume, more preferably 90% by volume or more, and even more preferably 95% by volume or more. The manufacturing method of the endothelin action inhibitor which performs 1 or more types of processes chosen from the group which consists of a low-temperature preservation | save process and a hydrolysis process with respect to the obtained extract.
＜４５＞前記アルコール水溶液がエタノール水溶液である、前記＜４４＞項記載の製造方法。
＜４６＞前記サルオガセ属地衣類がナガサルオガセ、ヨコワサルオガセ、フジサルオガセ、ニセフジサルオガセ及びクシノハサルオガセからなる群より選ばれる少なくとも１種、好ましくはナガサルオガセ及びヨコワサルオガセからなる群より選ばれる少なくとも１種である、前記＜４４＞又は＜４５＞項記載の製造方法。
＜４７＞前記アルコール水溶液を用いた抽出、前記加水処理、及び前記低温保存処理をこの順で行う、前記＜４４＞～＜４６＞のいずれか１項記載の製造方法。
＜４８＞前記アルコール水溶液を用いた抽出、前記低温保存処理、前記加水処理、及び前記低温保存処理をこの順で行う、前記＜４４＞～＜４６＞のいずれか１項記載の製造方法。
＜４９＞得られたエンドセリン作用抑制剤中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が２以上、好ましくは１０以上、である、前記＜４４＞～＜４８＞のいずれか１項記載の製造方法。 <45> The method according to <44>, wherein the aqueous alcohol solution is an aqueous ethanol solution.
<46> At least one selected from the group consisting of Nagasaga Ogase, Yokosawa Ogase, Fujisaru Ogase, Nisefujisaru Ogase, and Kushinohasa Ogase, preferably at least one selected from the group consisting of Nagasaro Ogase and Yokosawa Ogase The method according to <44> or <45>, wherein
<47> The production method according to any one of <44> to <46>, wherein the extraction using the aqueous alcohol solution, the hydrolysis treatment, and the low-temperature storage treatment are performed in this order.
<48> The method according to any one of <44> to <46>, wherein the extraction using the aqueous alcohol solution, the low-temperature storage treatment, the hydration treatment, and the low-temperature storage treatment are performed in this order.
<49> When the content of usnic acid in the obtained endothelin action inhibitor is set to 1 on a mass basis, the difractic acid content is 2 or more, preferably 10 or more, <44> to <48> The manufacturing method of any one of Claims.
＜５０＞前記＜１８＞～＜２４＞及び＜４４＞～＜４８＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類の抽出物又はエンドセリン作用抑制剤を有効成分とする、美白剤。 <50> A whitening agent comprising, as an active ingredient, an extract of Salogase lichen or an endothelin action inhibitor obtained by the production method according to any one of <18> to <24> and <44> to <48> .
＜５１＞エンドセリン作用抑制剤又は美白剤としての、前記＜１＞～＜２４＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物の使用。
＜５２＞エンドセリン作用抑制剤又は美白剤の製造のための、前記＜１＞～＜２４＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物の使用。
＜５３＞前記＜１＞～＜２４＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物を、エンドセリン作用抑制剤又は美白剤として使用する方法。
＜５４＞前記＜１＞～＜２４＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物を用いる、エンドセリン作用抑制方法。
＜５５＞前記＜１＞～＜２４＞のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物を用いる、美白方法。
＜５６＞メラノサイト内のカルシウムイオン濃度の上昇を抑制してエンドセリン作用を抑制する、前記＜５１＞～＜５５＞のいずれか１項記載の使用又は方法。
＜５７＞前記サルオガセ属地衣類抽出物を化粧料の形態で適用する、前記＜５１＞～＜５６＞のいずれか１項記載の使用又は方法。 <51> Use of an extract of Ligocephalus lichen obtained by the production method according to any one of <1> to <24> as an endothelin action inhibitor or a whitening agent.
<52> Use of an extract of Salogase lichen obtained by the production method according to any one of <1> to <24> for the production of an endothelin action inhibitor or a whitening agent.
<53> A method of using the Salgogase lichen extract obtained by the production method according to any one of <1> to <24> as an endothelin action inhibitor or a whitening agent.
<54> A method for suppressing endothelin action, which uses the Salgaraceae lichen extract obtained by the production method according to any one of <1> to <24>.
<55> A whitening method using the Salgogase lichen extract obtained by the production method according to any one of <1> to <24>.
<56> The use or method according to any one of <51> to <55>, wherein the increase in calcium ion concentration in melanocytes is suppressed to suppress endothelin action.
<57> The use or method according to any one of the above items <51> to <56>, wherein the extract of Lichenaceae lichen is applied in the form of a cosmetic.
＜５８＞美白剤としての、前記＜２７＞～＜４２＞及び＜４４＞～＜４９＞のいずれか１項記載の製造方法により得られるエンドセリン作用抑制剤の使用。
＜５９＞美白剤の製造のための、前記＜２７＞～＜４２＞及び＜４４＞～＜４９＞のいずれか１項記載の製造方法により得られるエンドセリン作用抑制剤の使用。
＜６０＞前記＜２７＞～＜４２＞及び＜４４＞～＜４９＞のいずれか１項記載の製造方法により得られるエンドセリン作用抑制剤を、美白剤として使用する方法。
＜６１＞前記＜２７＞～＜４２＞及び＜４４＞～＜４９＞のいずれか１項記載の製造方法により得られるエンドセリン作用抑制剤を用いる、美白方法。
＜６２＞メラノサイト内のカルシウムイオン濃度の上昇を抑制してエンドセリン作用を抑制する、前記＜５８＞～＜６１＞のいずれか１項記載の使用又は方法。
＜６３＞前記エンドセリン作用抑制剤を化粧料の形態で適用する、前記＜５８＞～＜６２＞のいずれか１項記載の使用又は方法。 <58> Use of an endothelin action inhibitor obtained by the production method according to any one of <27> to <42> and <44> to <49> as a whitening agent.
<59> Use of an endothelin action inhibitor obtained by the production method according to any one of <27> to <42> and <44> to <49> for the production of a whitening agent.
<60> A method of using the endothelin action inhibitor obtained by the production method of any one of <27> to <42> and <44> to <49> as a whitening agent.
<61> A whitening method using an endothelin action inhibitor obtained by the production method according to any one of <27> to <42> and <44> to <49>.
<62> The use or method according to any one of <58> to <61>, wherein the increase in calcium ion concentration in melanocytes is suppressed to suppress endothelin action.
<63> The use or method according to any one of <58> to <62>, wherein the endothelin action inhibitor is applied in the form of a cosmetic.
　以下、本発明を実施例に基づきさらに詳細に説明するが、本発明はこれに限定されるものではない。 Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited thereto.
参考例１　ナガサルオガセ抽出物の調製
　ナガサルオガセの地位体（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．６９ｇ（蒸発残分０．２９８（ｗ／ｖ）％）を得た。 Reference Example 1 Preparation of Nagasaga Ogase Extract 10 g of Nagasaga Ogase (obtained from Shinwa Bussan Co., Ltd.) and 200 mL of 50% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 7 days, followed by filtration, Nagasaga Ogase Extract 199 .69 g (evaporation residue 0.298 (w / v)%) was obtained.
参考例２　ヨコワサルオガセ抽出物１の調製
　ヨコワサルオガセの地位体（ハイケム社より入手）５ｇと、５０体積％エタノール水溶液１００ｍＬとを混合して、４０℃で８日間抽出後濾過し、ヨコワサルオガセ抽出物１（７８．２６ｇ）を得た（蒸発残分０．５６％）。 Reference Example 2 Preparation of Yokosawa Ogase Extract 1 5 g of Yokosawa Ogase standing (obtained from Hichem) was mixed with 100 mL of a 50% by volume ethanol aqueous solution, extracted at 40 ° C. for 8 days, filtered, and then extracted. 1 (78.26 g) was obtained (evaporation residue 0.56%).
参考例３　ヨコワサルオガセ抽出物２の調製
　ヨコワサルオガセの地位体（ハイケム社より入手）５ｇと、８０体積％1,3-ブタンジオール水溶液１００ｍＬとを混合して、４０℃で８日間抽出後濾過し、ヨコワサルオガセ抽出物２（８７．０７ｇ）を得た（蒸発残分０．１４％）。 Reference Example 3 Preparation of Yokosawa Ogase Extract 2 5 g of Yokosawa Ogase standing (obtained from Hichem) and 100 mL of 80 volume% 1,3-butanediol aqueous solution were mixed, extracted at 40 ° C. for 8 days and filtered. Yokowa Sarugase extract 2 (87.07 g) was obtained (evaporation residue 0.14%).
参考例４　ジャーマンカミツレ抽出物の調製
　ジャーマンカミツレ（和名：カミツレ、新和物産社より入手）の花部４０ｇと、５０体積％エタノール含有水溶液４００ｍＬとを混合して、室温で１４日間抽出後ろ過し、ジャーマンカミツレ抽出物（２２１ｍＬ）を得た（蒸発残分２．６３％）。 Reference Example 4 Preparation of German chamomile extract 40 g of flower portion of German chamomile (Japanese name: chamomile, obtained from Shinwa Bussan Co., Ltd.) and 400 mL of 50% ethanol-containing aqueous solution were mixed and extracted after 14 days at room temperature and filtered German chamomile extract (221 mL) was obtained (evaporation residue 2.63%).
参考例５　ジフラクタ酸の製造
　ナガサルオガセ（新和物産社より購入）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物８９１ｍＬ（固形分３．１０ｇ）を得た。この抽出物中の固形分５５．３ｍｇをＨＰＬＣで分画し、ジフラクタ酸５．１０ｍｇ（収率９．２２％）を得た。
　単離成分の構造解析は、ＮＭＲにより行った。単離成分のＮＭＲスペクトルデータが文献で報告されているジフラクタ酸のスペクトルデータ（Phytochemistry，1987，Vol．26，No．12，p．3181-3185参照）とほぼ一致したことから、単離された成分はジフラクタ酸であると同定した。なお、ＮＭＲによる構造解析の結果を表１に示す。 Reference Example 5 Production of difractic acid 50 g Nagasagahogase (purchased from Shinwa Bussan Co., Ltd.) and 1 L of 50% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 7 days, followed by filtration to 891 mL Nagasagaogase extract (solid content 3. 10 g) was obtained. The solid content of 55.3 mg in this extract was fractionated by HPLC to obtain 5.10 mg (yield: 9.22%) of difractic acid.
The structural analysis of the isolated component was performed by NMR. It was isolated because the NMR spectral data of the isolated component was almost identical to the spectral data of difractic acid reported in the literature (see Phytochemistry, 1987, Vol. 26, No. 12, p. 3181-3185) The component was identified as difractic acid. The results of structural analysis by NMR are shown in Table 1.
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003
試験例　エンドセリン作用抑制効果の検証
　ナガサルオガセ抽出物、ヨコワサルオガセ抽出物、ジャーマンカミツレ抽出物及びジフラクタ酸について、下記の評価系を用いてエンドセリン作用抑制効果を検証した。なお、エンドセリンはメラノサイトに作用してカルシウム濃度を上昇させ、メラニン産生を増加させることが知られており（Yada　Y．et　al.，J．Biol．Chem.，1991，vol．266，p．18352-18357；Imokawa　G．et　al.，J．Biol．Chem.，1992，vol．267，p．24675-24680など参照）、エンドセリンの当該作用を抑制する物質は美白成分として有用である。 Test example Verification of endothelin action inhibitory effect About Nagasaru Ogase extract, Yokosawa Ogase extract, German chamomile extract and difractic acid, the endothelin action inhibitory effect was verified using the following evaluation system. Endothelin is known to act on melanocytes to increase calcium concentration and increase melanin production (Yada Y. et al., J. Biol. Chem., 1991, vol. 266, p. 18352). -18357; Imokawa G. et al., J. Biol. Chem., 1992, vol. 267, p. 24675-24680, etc.), substances that suppress the action of endothelin are useful as whitening ingredients.
　正常ヒト新生児表皮由来メラノサイト（ＮＨＥＭｓ；クラボウ社製）を、蛍光検出用の９６穴プレートに３×１０4ｃｅｌｌｓ／ｗｅｌｌ（２００μＬ／ｗｅｌｌ）で播種し、５％ＣＯ2下にて３７℃で培養した。培地には、ＰＭＡ（－）の増殖用添加剤（ＨＭＧＳ）を含むＭｅｄｉｕｍ　２５４を用いた。
　３日間培養後、培養プレートからデカントで培地を除去し、細胞内Ｃａ2++測定試薬Fluo4-AMを含むアッセイバッファーに置換し、３７℃で１時間インキュベートした。次に、ＦＤＳＳ（Functional　Drug　Screening　System）機器において測定開始２０秒後に所定濃度のジフラクタ酸のエタノール溶液、ナガサルオガセ抽出物のエタノール溶液、ヨコワサルオガセ抽出物のジメチルスルホキシド溶液又はジャーマンカミツレ抽出物のエタノール溶液で１分間前処理した後、リガンドであるエンドセリン（ＥＴ－１、終濃度１００ｎＭ）を添加し、Fluo4-AMの蛍光をＥｘ．４８０ｎｍ／Ｅｍ．５４０ｎｍの測定波長で測定開始から５分後まで経時的に検出した。
　コントロールにおけるFluo4-AMの蛍光増加比（Max．ratio-Min．ratio）を１００とした場合の相対値（％）で、各化合物又は抽出物における、エンドセリン刺激による細胞内カルシウム（カルシウムイオン）濃度上昇率（％）を算出し、エンドセリン作用抑制効果を評価した。なお、コントロールとして、５０体積％エタノール水溶液又はジメチルスルホキシドを添加したものを用いた。
　その結果を表２に示す。 Normal human neonatal epidermis-derived melanocytes (NHEMs; manufactured by Kurabo Industries) are seeded in 96-well plates for fluorescence detection at 3 × 10 4 cells / well (200 μL / well) and cultured at 37 ° C. under 5% CO 2. did. As the medium, Medium 254 containing PMA (-) growth additive (HMGS) was used.
After culturing for 3 days, the medium was removed from the culture plate by decantation, replaced with assay buffer containing intracellular Ca 2 ++ measurement reagent Fluo4-AM, and incubated at 37 ° C. for 1 hour. Next, 20 seconds after the start of the measurement in an FDSS (Functional Drug Screening System) device, an ethanol solution of difractic acid at a predetermined concentration, an ethanol solution of Nagasaru Ogase extract, a dimethyl sulfoxide solution of Yokosawa Ogase extract or an ethanol solution of German chamomile extract And pre-treatment for 1 minute, followed by addition of the ligand endothelin (ET-1, final concentration of 100 nM) and fluorescence of Fluo4-AM with Ex. 480 nm / Em. Detection was performed over time from the start of measurement at a measurement wavelength of 540 nm until 5 minutes later.
Relative value (%) with Fluo4-AM fluorescence increase ratio (Max.ratio-Min.ratio) in control as 100, increase in intracellular calcium (calcium ion) concentration by endothelin stimulation in each compound or extract The rate (%) was calculated, and the endothelin action inhibitory effect was evaluated. As a control, a 50% by volume ethanol aqueous solution or dimethyl sulfoxide added was used.
The results are shown in Table 2.
Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004
　ジャーマンカミツレ抽出物は、エンドセリンの作用によるメラノサイトのＣａ濃度上昇を抑制し、メラニン産生を抑制することが知られている（例えば、Pigment　Cell　Res.，1997，vol．10，p．218-228参照）。表２から明らかなように、ジャーマンカミツレ抽出物を添加した系では、濃度依存的なカルシウム濃度上昇抑制作用が確認された。特に濃度が１ｖ／ｖ％の系では、２０％以上もカルシウム濃度上昇が抑制され、優れたエンドセリン抑制作用が確認された。
　これに対して、表２から明らかなように、ナガサルオガセ抽出物、ヨコワサルオガセ抽出物又はジフラクタ酸を添加した系では、エンドセリン刺激によるメラノサイト内のカルシウム濃度上昇が抑制された。すなわち、ナガサルオガセ及びヨコワサルオガセなどのサルオガセ属地衣類の抽出物、並びにジフラクタ酸は、公知の美白成分であるジャーマンカミツレ抽出物と同等またはそれ以上の、優れたエンドセリン抑制作用を有し、美白成分として有用であることがわかる。 German chamomile extract is known to suppress the increase in Ca concentration of melanocytes by the action of endothelin and suppress melanin production (see, for example, Pigment Cell Res., 1997, vol. 10, p. 218-228). ). As is clear from Table 2, in the system to which the German chamomile extract was added, a concentration-dependent calcium concentration increase inhibitory action was confirmed. In particular, in a system with a concentration of 1 v / v%, an increase in calcium concentration was suppressed by 20% or more, and an excellent endothelin inhibitory action was confirmed.
On the other hand, as is clear from Table 2, in the system to which Nagasaga Ogase extract, Yokosawa Ogase extract or difractic acid was added, an increase in calcium concentration in melanocytes by endothelin stimulation was suppressed. That is, extracts of lichen genus lichens such as Nagasaru Ogase and Yokosawa Ogase, and difractic acid have an excellent endothelin inhibitory effect equal to or higher than that of German chamomile extract, which is a known whitening component, and as a whitening component It turns out that it is useful.
（第１の実施態様に関する実施例及び比較例）
　以下、第１の実施態様に関する実施例及び比較例について説明する。 (Examples and comparative examples relating to the first embodiment)
Examples and comparative examples relating to the first embodiment will be described below.
実施例１－１
　ナガサルオガセ（新和物産社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０７．３５ｇ（蒸発残分０．２０２（ｗ／ｖ）％）を得た。 Example 1-1
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, 207.35 g of Nagasaru Ogase extract (evaporation residue 0.202 (w / w v)%).
実施例１－２
　ナガサルオガセ（新和物産社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０６．５９ｇ（蒸発残分０．２６６（ｗ／ｖ）％）を得た。 Example 1-2
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed and extracted at 30 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 206.59 g (evaporation residue 0.266 (w / v)%).
実施例１－３
　ナガサルオガセ（新和物産社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０８．２６ｇ（蒸発残分０．２８０（ｗ／ｖ）％）を得た。 Example 1-3
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed, extracted at 40 ° C. for 7 days, filtered, and Nagasal Ogase extract 208.26 g (evaporation residue 0.280 (w / w v)%).
実施例１－４
　ナガサルオガセ（新和物産社より入手）１０ｇと２０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０１．４９ｇ（蒸発残分０．１７２（ｗ／ｖ）％）を得た。 Example 1-4
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 20 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.49 g (evaporation residue 0.172 (w / v)%).
実施例１－５
　ナガサルオガセ（新和物産社より入手）１０ｇと３０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．９０ｇ（蒸発残分０．１４６（ｗ／ｖ）％）を得た。 Example 1-5
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 30 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 199.90 g (evaporation residue 0.146 (w / v)%).
実施例１－６
　ナガサルオガセ（新和物産社より入手）１０ｇと４０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２００．７０ｇ（蒸発残分０．１９０（ｗ／ｖ）％）を得た。 Example 1-6
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 40 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasal Ogase extract 200.70 g (evaporation residue 0.190 (w / v)%).
実施例１－７
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０１．６７ｇ（蒸発残分０．１７８（ｗ／ｖ）％）を得た。 Example 1-7
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 5 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.67 g (evaporation residue 0.178 (w / v)%).
実施例１－８
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して１５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０２．５７ｇ（蒸発残分０．１９２（ｗ／ｖ）％）を得た。 Example 1-8
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 15 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 202.57 g (evaporation residue 0.192 (w / v)%).
実施例１－９
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．６９ｇ（蒸発残分０．２９８（ｗ／ｖ）％）を得た。 Example 1-9
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.69 g (evaporation residue 0.298 (w / v)%).
実施例１－１０
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２００．５４ｇ（蒸発残分０．２５８（ｗ／ｖ）％）を得た。 Example 1-10
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 30 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 200.54 g (evaporation residue 0.258 (w / v)%).
実施例１－１１
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．４９ｇ（蒸発残分０．３１６（ｗ／ｖ）％）を得た。 Example 1-11
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.49 g (evaporation residue 0.316 (w / v)%).
実施例１－１２
　ナガサルオガセ（新和物産社より入手）１０ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９８．６９ｇ（蒸発残分０．２４２（ｗ／ｖ）％）を得た。 Example 1-12
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 60 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 198.69 g (evaporation residue 0.242 (w / v)%).
実施例１－１３
　ナガサルオガセ（新和物産社より入手）１０ｇと７０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９５．１ｇ（蒸発残分０．２３２（ｗ／ｖ）％）を得た。 Example 1-13
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 70 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, 195.1 g of Nagasal Ogase extract (evaporation residue 0.232 (w / v)%).
実施例１－１４
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８５．６ｇ（蒸発残分０．２１６（ｗ／ｖ）％）を得た。 Example 1-14
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 80 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.6 g (evaporation residue 0.216 (w / v)%).
比較例１－１
　ナガサルオガセ（新和物産社より入手）１０ｇと９０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８３．４ｇ（蒸発残分０．１９２（ｗ／ｖ）％）を得た。 Comparative Example 1-1
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 90 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaga Ogase extract 183.4 g (evaporation residue 0.192 (w / v)%).
比較例１－２
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８４．２４ｇ（蒸発残分０．１２８（ｗ／ｖ）％）を得た。 Comparative Example 1-2
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 5 ° C. for 7 days, filtered, and Nagasaru Ogase extract 184.24 g (evaporation residue 0.128 (w / v)%).
比較例１－３
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して１５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８４．０ｇ（蒸発残分０．１５４（ｗ／ｖ）％）を得た。 Comparative Example 1-3
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% aqueous ethanol solution 200 mL were mixed and extracted at 15 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 184.0 g (evaporation residue 0.154 (w / v)%).
比較例１－４
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８４．２２ｇ（蒸発残分０．１７４（ｗ／ｖ）％）を得た。 Comparative Example 1-4
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% aqueous ethanol solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 184.22 g (evaporation residue 0.174 (w / v)%).
比較例１－５
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８６．４２ｇ（蒸発残分０．２１８（ｗ／ｖ）％）を得た。 Comparative Example 1-5
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 30 ° C. for 7 days, filtered, 186.42 g of Nagasal Ogase extract (evaporation residue 0.218 (w / v)%).
比較例１－６
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８５．０６ｇ（蒸発残分０．２６６（ｗ／ｖ）％）を得た。 Comparative Example 1-6
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 40 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.06 g (evaporation residue 0.266 (w / v)%).
比較例１－７
　ナガサルオガセ（新和物産社より入手）２５ｇと９９．５体積％エタノール水溶液５００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物３６８．０ｇ（蒸発残分０．１５８（ｗ／ｖ）％）を得た。 Comparative Example 1-7
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 25 g and 99.5 vol% ethanol aqueous solution 500 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 368.0 g (evaporation residue 0.158 ( w / v)%).
ウスニン酸含有量の測定
　上記実施例１－１～１－１４及び比較例１－１～１－７で得られた各ナガサルオガセ抽出物中のウスニン酸含有量を、ＨＰＬＣを用いて下記の条件で分析した。測定に用いた装置及び測定条件は下記のとおりである。
 
測定機器(HPLC)：Agilent　Technologies　Open　LAB　CDS　LC02-1260、オートサンプラーG1329B　1260　ALS、カラムオーブンG1316A　1260　TCC、検出器G1315D　1260　DAD　VL、ポンプG1312B　1260　Bin　pump、G1379B　1260　μ-Degasser
 
＜HPLC分析条件＞
流速：０．７５ｍＬ/ｍｉｎ
使用したカラム：Inertsil　ODS-3　５μｍ(GL　Science)、3.0I.D.×１５０ｍｍ
検出波長：２５４ｎｍ
カラム温度：４０℃
移動相：
Ａ：0.1%トリフルオロ酢酸含有水溶液、Ｂ：0.1%トリフルオロ酢酸含有メタノール
　　　０～１０分：70%Ｂ
　　１０～１５分：70％Ｂ→100%Ｂ
　　１５～２０分：100％Ｂ
 
　結果を表３に示す。 Measurement of usnic acid content The content of usnic acid in each Nagasaruogase extract obtained in Examples 1-1 to 1-14 and Comparative Examples 1-1 to 1-7 was measured under the following conditions using HPLC. Analyzed with The apparatus and measurement conditions used for the measurement are as follows.

Measuring instruments (HPLC): Agilent Technologies Open LAB CDS LC02-1260, Autosampler G1329B 1260 ALS, Column oven G1316A 1260 TCC, Detector G1315D 1260 DAD VL, Pump G1312B 1260 Bin pump, G1379B 1260 μ-Degasser

<HPLC analysis conditions>
Flow rate: 0.75 mL / min
Column used: Inertsil ODS-3 5 μm (GL Science), 3.0 ID × 150 mm
Detection wavelength: 254 nm
Column temperature: 40 ° C
Mobile phase:
A: 0.1% trifluoroacetic acid-containing aqueous solution, B: 0.1% trifluoroacetic acid-containing methanol 0 to 10 minutes: 70% B
10-15 minutes: 70% B → 100% B
15-20 minutes: 100% B

The results are shown in Table 3.
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000005
　表３の結果から明らかなように、本発明の第１の実施態様の方法によれば、ウスニン酸の含有量が十分に低いナガサルオガセ抽出物を得ることができる。
　これに対して、アルコール含有率の高すぎる抽出溶媒を用いてナガサルオガセを抽出すると、ウスニン酸含有量が上昇した（比較例１－１～１－７）。 As is apparent from the results in Table 3, according to the method of the first embodiment of the present invention, a Nagasaruogase extract having a sufficiently low usnic acid content can be obtained.
On the other hand, when Nagasaruogase was extracted using an extraction solvent having an excessively high alcohol content, the usnic acid content increased (Comparative Examples 1-1 to 1-7).
　以上のように、サルオガセ属地衣類をアルコール含有率が８０体積％以下のアルコール水溶液を用いて抽出することで、ウスニン酸低含有のサルオガセ属地衣類抽出物を製造することができる。ウスニン酸は肝毒性を有するので、本発明の第１の実施態様によれば、安全性に優れ、エンドセリン作用抑制剤等の皮膚外用剤に好適に用いることができるサルオガセ属地衣類抽出物を製造することができる。 As described above, a Salusose lichen extract containing a low amount of usnic acid can be produced by extracting the Ligosa lichen using an aqueous alcohol solution having an alcohol content of 80% by volume or less. Since usnic acid has hepatotoxicity, according to the first embodiment of the present invention, an extract of Sargogase lichen that is excellent in safety and can be suitably used for an external skin preparation such as an endothelin action inhibitor is produced. be able to.
実施例１－１４
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で８日間抽出後ろ過し、ナガサルオガセ抽出物８５８．５ｇ（蒸発残分０．３７４（ｗ／ｖ）％）を得た。
　得られた抽出物を－５℃で１２日間保存後ろ過し、低温保存処理後のナガサルオガセ抽出物８３３．４ｇ（蒸発残分０．３６０（ｗ／ｖ）％）を得た。
　低温保存処理前後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表４に示す。 Example 1-14
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasaga Ogase extract 858.5 g (evaporation residue 0.374 (w / v)%).
The obtained extract was stored at −5 ° C. for 12 days and then filtered to obtain 833.4 g of Nagasaroogase extract (low evaporation residue 0.360 (w / v)%) after the low-temperature storage treatment.
The Usunic acid content was measured by the same method as above for the Nagasaga Ogase extract before and after the low-temperature storage treatment. The results are shown in Table 4.
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000006
　表４の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理を行なうことにより、ウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Table 4, the Ussanoic acid content is further reduced by performing low-temperature storage treatment on the Nagasaruogase extract obtained using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent. can do.
実施例１－１６
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で８日間抽出後ろ過し、ナガサルオガセ抽出物８５２．８ｇ（蒸発残分０．２６８（ｗ／ｖ）％）を得た。得られた抽出物を－５℃で１３日間保存後、ろ過することにより、ナガサルオガセ抽出物処理前品８３７．２ｇ（蒸発残分０．２５６（ｗ／ｖ）％）を得た。
　次に、ナガサルオガセ抽出物処理前品１０ｍＬと蒸留水２．５ｍＬとを混合してエタノールの体積が４０％となるよう希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。さらに、加水処理後のナガサルオガセ抽出物を－５℃で４日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１１．１ｇ（蒸発残分０．１３０（ｗ／ｖ）％）を得た。
　ナガサルオガセ抽出物処理前品、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表５に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表５に示す。 Example 1-16
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasal Ogase extract 852.8 g (evaporation residue 0.268 (w / w v)%). The obtained extract was stored at −5 ° C. for 13 days and filtered to obtain 837.2 g (evaporation residue 0.256 (w / v)%) of the Nagasaga Ogase extract before treatment.
Next, 10 mL of Nagasaga Ogase extract pretreatment product and 2.5 mL of distilled water were mixed and diluted so that the volume of ethanol was 40%. . Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 4 days and then filtered, and the Nagasal Ogase extract after the hydrous / low temperature preservation treatment was 11.1 g (evaporation residue 0.130 (w / v)%). Got.
Usasic acid content was measured by the same method as above for the Nagasaga Ogase extract before treatment, the Nagasaga Ogase extract after the hydrotreatment, and the Nagasal Ogase extract after the hydrothermal treatment at low temperature. The results are shown in Table 5. In addition, the theoretical value of the usnic acid content contained in the Nagasaru Ogase extract after the hydrotreatment is also shown in Table 5.
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000007
　表５の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理、加水処理及び低温保存処理をこの順で行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is clear from the results in Table 5, a low-temperature storage treatment, a hydration treatment, and a low-temperature storage treatment are performed in this order for the Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent. Thus, the usnic acid content contained in the Nagasaru Ogase extract can be further reduced.
実施例１－１７
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物８３８．５ｇ（蒸発残分０．２１８（ｗ／ｖ）％）を得た。
　次に、加水・低温保存処理前のナガサルオガセ抽出物８００ｍＬと蒸留水２００ｍＬとを混合してエタノールの体積が４０％となるよう希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。さらに、加水処理後のナガサルオガセ抽出物を－５℃で７日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物９１６．２ｇ（蒸発残分０．１３０（ｗ／ｖ）％）を得た。
　加水・低温保存処理前のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表６に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表６に示す。 Example 1-17
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 1 L of 50% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 7 days, filtered, and 838.5 g (Evaporation residue) Min 0.218 (w / v)%).
Next, 800 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 200 mL of distilled water are mixed and diluted so that the volume of ethanol becomes 40%. Obtained. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 7 days and filtered, and 916.2 g of the Nagasal Ogase extract after the hydrothermal treatment at low temperature (Evaporation residue 0.130 (w / v)%) Got.
Usasic acid content was measured by the same method as described above for the Nagasaga Ogase extract before the hydrolysis / low temperature preservation treatment and the Nagasal Ogase extract after the hydrolysis / low temperature preservation treatment. The results are shown in Table 6. In addition, the theoretical value of the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment is also shown in Table 6.
Figure JPOXMLDOC01-appb-T000008
Figure JPOXMLDOC01-appb-T000008
　表６の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理及び低温保存処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results of Table 6, Nagasaga Ogase extract is obtained by subjecting Nagasaga Ogase extract obtained by using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent to water treatment and low-temperature storage treatment. Can further reduce the usnic acid content.
実施例１－１８
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）０．６５ｍｇ（抽出物中固形分に対して１０重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭１０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．０８（ｗ／ｖ）％））。その結果を表７に示す。 Example 1-18
5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. Then, add 0.65 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (10% by weight based on the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 10 wt% -treated product was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment. The acid content was measured (evaporation residue 0.08 (w / v)%)). The results are shown in Table 7.
実施例１－１９
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）３．２５ｍｇ（抽出物中固形分に対して５０重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表７に示す。 Example 1-19
5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. After dilution, add 3.25 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (50% by weight based on the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 50 wt% treated product was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment. The acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
実施例１－２０
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）６．５０ｍｇ（抽出物中固形分に対して１００重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭１００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表７に示す。 Example 1-20
5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. After dilution, add 6.50 mg of activated carbon (trade name: Shirahige M, manufactured by Nippon Enviro Chemicals) (100% by weight with respect to the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 100 wt% treated product was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment. The acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
実施例１－２１
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）９．７５ｍｇ（抽出物中固形分に対して１５０重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭１５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表７に示す。 Example 1-21
5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. Then, 9.75 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (150% by weight with respect to the solid content in the extract) is added, and the mixture is stirred at room temperature for 45 minutes and then filtered. Thus, a product after treatment with 150 wt% of activated carbon was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment. The acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
実施例１－２２
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１３．０ｍｇ（抽出物中固形分に対して２００重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭２００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表７に示す。 Example 1-22
5 mL of Nagasaru Ogase extract (Nagasaru Ogase extract before activated carbon treatment) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment (solid content 6.5 mg) and 99.5 vol% ethanol aqueous solution 45 mL were mixed. 13.0 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (200% by weight with respect to the solid content in the extract) is added, stirred for 45 minutes at room temperature, and then filtered. Thus, a product after treatment with 200 wt% of activated carbon was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as that of the product before dilution at the time of activated carbon treatment. The acid content was measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 7.
Figure JPOXMLDOC01-appb-T000009
Figure JPOXMLDOC01-appb-T000009
　表７の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理、低温保存処理及び活性炭処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Table 7, Nagasagaogase extract obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and activated carbon treatment. The usnic acid content contained in the Saruogase extract can be further reduced.
実施例１－２３
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物（ヘキサン洗浄処理前抽出物）５０ｍＬにヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄１回品）、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した。その結果を表８に示す。 Example 1-23
50 mL of hexane and 5 mL of 99.5 vol% ethanol aqueous solution were added to 50 mL of Nagasaroogase extract (extract before hexane washing) obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment, and a separating funnel was used. After separating into two layers, the lower layer was recovered (hexane washed once product), and the usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 8.
実施例１－２４
　実施例１－２３で得られたナガサルオガセ抽出物５０ｍＬに、再度ヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄２回品）、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した。その結果を表８に示す。 Example 1-24
To 50 mL of Nagasaga Ogase extract obtained in Example 1-23, hexane 50 mL and 99.5% by volume aqueous ethanol solution 5 mL were added again, and after separating into two layers using a separatory funnel, the lower layer was recovered (hexane The usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 8.
実施例１－２５
　実施例１－２４で得られたナガサルオガセ抽出物４５ｍＬに、再度ヘキサン４５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄３回品）、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した。その結果を表８に示す。 Example 1-25
Add 45 mL of hexane again to 45 mL of Nagasaga Ogase extract obtained in Example 1-24, separate into two layers using a separatory funnel, and recover the lower layer (hexane washed three times), as above The usnic acid content was measured by HPLC according to the above method. The results are shown in Table 8.
Figure JPOXMLDOC01-appb-T000010
Figure JPOXMLDOC01-appb-T000010
　表８の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理、低温保存処理及びヘキサン洗浄処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Table 8, the Nagasaruogase extract obtained using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and hexane washing treatment. The usnic acid content contained in the Nagasaru Ogase extract can be further reduced.
実施例１－２６
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で３日間抽出後ろ過し、ナガサルオガセ抽出物８５８．３ｇ（蒸発残分０．４６０（ｗ／ｖ）％）を得た。
　得られたナガサルオガセ抽出物を－５℃で１２日間保存後、ろ過することにより活性炭処理前のナガサルオガセ抽出物８４９．２ｇ（蒸発残分０．３６０（ｗ／ｖ）％）を得た。
　得られた活性炭処理前のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表９に示す。 Example 1-26
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% aqueous ethanol solution 1 L were mixed and extracted at 20 ° C. for 3 days followed by filtration. Nagasaru Ogase extract 858.3 g (evaporation residue 0.460 (w / v)%).
The obtained Nagasaga Ogase extract was stored at −5 ° C. for 12 days and then filtered to obtain 849.2 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before activated carbon treatment.
About the Nagasaruogase extract before the activated carbon treatment obtained, the usnic acid content was measured by the same method as described above. The results are shown in Table 9.
実施例１－２７
　実施例１－２６で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１．０８ｍｇ（抽出物中固形分に対して１０重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭１０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．２８０（ｗ／ｖ）％））。その結果を表９に示す。 Example 1-27
After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) 1.08 mg (manufactured by Enviro Chemicals) (10% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with 10 wt% of activated carbon. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid content was measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 9.
実施例１－２８
　実施例１－２６で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）５．４０ｍｇ（抽出物中固形分に対して５０重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．２８０（ｗ／ｖ）％））。その結果を表９に示す。 Example 1-28
After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 5.40 mg (50 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon 50 wt%. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid content was measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 9.
実施例１－２９
　実施例１－２６で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１０．８０ｍｇ（抽出物中固形分に対して１００重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭１００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．２７０（ｗ／ｖ）％））。その結果を表９に示す。 Example 1-29
After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 10.80 mg (100% by weight based on solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with activated carbon 100 wt%. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid content was measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 9.
実施例１－３０
　実施例１－２６で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１６．２０ｍｇ（抽出物中固形分に対して１５０重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭１５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．２７０（ｗ／ｖ）％））。その結果を表９に示す。 Example 1-30
After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 16.20 mg (150 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with 150 wt% of activated carbon. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid content was measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 9.
実施例１－３１
　実施例１－２６で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）２１．６０ｍｇ（抽出物中固形分に対して２００重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭２００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．２７０（ｗ／ｖ）％））。その結果を表９に示す。 Example 1-31
After adding and diluting 27 mL of 99.5 vol% ethanol aqueous solution to 3 mL of Nagasaruogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 1-26, activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd. (21.60 mg) (200% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon of 200 wt%. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid content was measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 9.
Figure JPOXMLDOC01-appb-T000011
Figure JPOXMLDOC01-appb-T000011
　表９の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理及び活性炭処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Table 9, Nagasaga Ogase extract is obtained by performing low-temperature storage treatment and activated carbon treatment on Nagasaga Ogase extract obtained using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent. Can further reduce the usnic acid content.
実施例１－３２
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で８日間抽出後ろ過し、ナガサルオガセ抽出物８５５．８ｇ（蒸発残分０．２７０（ｗ／ｖ）％）を得た。
　得られたナガサルオガセ抽出物を－５℃で１３日間保存後、ろ過することによりヘキサン洗浄処理前のナガサルオガセ抽出物８３９．５ｇ（蒸発残分０．３６０（ｗ／ｖ）％）を得た。
　得られたヘキサン洗浄処理前のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表１０に示す。 Example 1-32
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed and extracted at 20 ° C. for 8 days followed by filtration. Nagasaru Ogase extract 855.8 g (evaporation residue 0.270 (w / v)%).
The obtained Nagasaga Ogase extract was stored at −5 ° C. for 13 days and then filtered to obtain 839.5 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before hexane washing treatment. .
About the Nagasagaogase extract before the obtained hexane washing process, the usnic acid content was measured by the method similar to the above. The results are shown in Table 10.
実施例１－３３
　実施例１－３２で得られたヘキサン洗浄処理前のナガサルオガセ抽出物５０ｍＬにヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄１回品）、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した。その結果を表１０に示す。 Example 1-33
Add 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution to 50 mL of Nagasaru Ogase extract obtained in Example 1-32 before washing with hexane, separate into two layers using a separatory funnel, and recover the lower layer Then, the usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 10.
実施例１－３４
　実施例１－３３で得られたナガサルオガセ抽出物５０ｍＬに、再度ヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄２回品）、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した。その結果を表１０に示す。 Example 1-34
To 50 mL of Nagasaga Ogase extract obtained in Example 1-33, 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution were added again. After separating into two layers using a separatory funnel, the lower layer was recovered (hexane The usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 10.
実施例１－３５
　実施例１－３４で得られたナガサルオガセ抽出物５０ｍＬに、再度ヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄３回品）、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した（蒸発残分０．２８６（ｗ／ｖ）％）。その結果を表１０に示す。 Example 1-35
To 50 mL of Nagasaga Ogase extract obtained in Example 1-34, 50 mL of hexane and 5 mL of a 99.5% by volume ethanol aqueous solution were added again. After separating into two layers using a separatory funnel, the lower layer was recovered (hexane The product was washed three times), and the usnic acid content was measured by HPLC in the same manner as described above (evaporation residue 0.286 (w / v)%). The results are shown in Table 10.
Figure JPOXMLDOC01-appb-T000012
Figure JPOXMLDOC01-appb-T000012
　表１０の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理及びヘキサン洗浄処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Table 10, Nagasagaogase extract is obtained by performing low-temperature storage treatment and hexane washing treatment on Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent. The usnic acid content contained in the product can be further reduced.
実施例１－３６～１－４３
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物１０ｍＬを６０℃で静置保存し、表１１に示した各保存時間後のナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでウスニン酸の含有量を測定した。その結果を表１１に示す。 Examples 1-36 to 1-43
10 mL of the Nagasaga Ogase extract after the hydrolyzing and low-temperature storage treatment obtained in Example 1-17 was stored at 60 ° C., and the Nagasaga Ogase extract after each storage time shown in Table 11 was the same as described above. The usnic acid content was measured by HPLC according to the method. The results are shown in Table 11.
Figure JPOXMLDOC01-appb-T000013
Figure JPOXMLDOC01-appb-T000013
実施例１－４４～１－４７
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物１５０ｍＬを７０℃で表１１に示した時間で攪拌を行い、得られたナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでウスニン酸の含有量を測定した。その結果を表１２に示す。 Examples 1-44 to 1-47
The same method as described above for the Nagasaga Ogase extract obtained by stirring 150 mL of Nagasaro Ogase extract obtained in Example 1-17 after the hydrolyzing and low-temperature storage treatment at 70 ° C. for the time shown in Table 11. Was used to measure the content of usnic acid by HPLC. The results are shown in Table 12.
Figure JPOXMLDOC01-appb-T000014
Figure JPOXMLDOC01-appb-T000014
実施例１－４８～１－５０
　実施例１－１７で得られた加水・低温保存処理後のナガサルオガセ抽出物１５０ｍＬを８０℃で表１２に示した時間で攪拌を行い、得られたナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでウスニン酸の含有量を測定した。その結果を表１３に示す。 Examples 1-48 to 1-50
150 mL of the Nagasaga Ogase extract obtained after the hydrolyzing and low-temperature storage treatment obtained in Example 1-17 was stirred at 80 ° C. for the time shown in Table 12, and the obtained Nagasaga Ogase extract was subjected to the same method as described above. Was used to measure the content of usnic acid by HPLC. The results are shown in Table 13.
Figure JPOXMLDOC01-appb-T000015
Figure JPOXMLDOC01-appb-T000015
　表１１～１３の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理、低温保存処理及び加熱処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Tables 11 to 13, Nagasagaogase extract obtained by using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and heat treatment. The usnic acid content contained in the Nagasaru Ogase extract can be further reduced.
実施例１－５１
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７５．６２ｇ（蒸発残分０．２９０（ｗ／ｖ）％）を得た。 Example 1-51
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaga Ogase extract 175.62 g (evaporation residue 0 .290 (w / v)%).
実施例１－５２
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７４．２５ｇ（蒸発残分０．３４０（ｗ／ｖ）％）を得た。 Example 1-52
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and 174.25 g of Nagasal Ogase extract (evaporation residue 0 .340 (w / v)%).
実施例１－５３
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７７．５７ｇ（蒸発残分０．４１０（ｗ／ｖ）％）を得た。 Example 1-53
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 177.57 g (evaporation residue 0 .410 (w / v)%).
実施例１－５４
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７８．１９ｇ（蒸発残分０．２６０（ｗ／ｖ）％）を得た。 Example 1-54
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaga Ogase extract 178.19 g (evaporation residue 0 260 (w / v)%).
実施例１－５５
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７９．８２ｇ（蒸発残分０．３００（ｗ／ｖ）％）を得た。 Example 1-55
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and Nagasal Ogase extract 179.82 g (evaporation residue 0 .300 (w / v)%).
実施例１－５６
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８２．１４ｇ（蒸発残分０．３８０（ｗ／ｖ）％）を得た。 Example 1-56
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 182.14 g (evaporation residue 0 .380 (w / v)%).
比較例１－８
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８２．５５ｇ（蒸発残分０．１２（ｗ／ｖ）％）を得た。 Comparative Example 1-8
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% 1,3-butanediol aqueous solution 200 mL were mixed, extracted at 25 ° C. for 7 days, filtered, and Nagasaga Ogase extract 182.55 g (evaporation residue 0 .12 (w / v)%).
ウスニン酸含有量の測定
　上記実施例１－５１～１－５６及び比較例１－８で得られた各ナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した。その結果を表１４に示す。 Measurement of usnic acid content For each of Nagasagaogase extracts obtained in Examples 1-51 to 1-56 and Comparative Example 1-8, the usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 14.
Figure JPOXMLDOC01-appb-T000016
Figure JPOXMLDOC01-appb-T000016
　表１４の結果から明らかなように、本発明の第１の実施態様の方法によれば、ウスニン酸の濃度が十分に低いナガサルオガセ抽出物を得ることができる。
　これに対して、アルコール含有率の高すぎる抽出溶媒を用いてナガサルオガセを抽出すると、ウスニン酸含有量が上昇した（比較例１－８）。
　以上のように、サルオガセ属地衣類をアルコール含有率が８０体積％以下のアルコール水溶液を用いて抽出することで、ウスニン酸低含有のサルオガセ属地衣類抽出物を製造することができる。ウスニン酸は肝毒性を有するので、本発明の第１の実施態様によれば、安全性に優れ、エンドセリン作用抑制剤等の皮膚外用剤に好適に用いることができるナガサルオガセ抽出物を製造することができる。 As is clear from the results in Table 14, according to the method of the first embodiment of the present invention, a Nagasaruogase extract having a sufficiently low concentration of usnic acid can be obtained.
On the other hand, when Nagasagaogase was extracted using an extraction solvent with an excessively high alcohol content, the usnic acid content increased (Comparative Example 1-8).
As described above, a Salusose lichen extract containing a low amount of usnic acid can be produced by extracting the Ligosa lichen using an alcohol aqueous solution having an alcohol content of 80% by volume or less. Since usnic acid has hepatotoxicity, according to the first embodiment of the present invention, an extract of Nagasaga Ogase which is excellent in safety and can be suitably used for an external skin preparation such as an endothelin action inhibitor is produced. Can do.
実施例１－５７
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７７．６ｇ（蒸発残分０．４２０（ｗ／ｖ）％）を得た。
　得られた抽出物１０ｍＬを－５℃で５日間保存後ろ過し、低温保存処理後のナガサルオガセ抽出物８．９ｇ（蒸発残分０．３８０（ｗ／ｖ）％）を得た。 Example 1-57
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract (177.6 g) 420 (w / v)%).
10 mL of the obtained extract was stored at −5 ° C. for 5 days and then filtered to obtain 8.9 g of Nagasaga Ogase extract after evaporation at low temperature (evaporation residue 0.380 (w / v)%).
　低温保存処理前後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表１５に示す。 For the Nagasaga Ogase extract before and after the low-temperature storage treatment, the usnic acid content was measured by the same method as described above. The results are shown in Table 15.
Figure JPOXMLDOC01-appb-T000017
Figure JPOXMLDOC01-appb-T000017
　表１５の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理を行なうことにより、ウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Table 15, the Ussanoic acid content is further reduced by performing a low-temperature storage treatment on the Nagasagaogase extract obtained using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent. can do.
実施例１－５８
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物１７７．６ｇ（蒸発残分０．４２０（ｗ／ｖ）％）を得た。
　次に、加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水１ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物９．５ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物８．６ｇ（蒸発残分０．３５０（ｗ／ｖ）％）を得た。 Example 1-58
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% 1,3-butanediol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration. 0.6 g (evaporation residue 0.420 (w / v)%) was obtained.
Next, the mixture was diluted by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 1 mL of distilled water to perform starch, and a Nagasal Ogase extract after hydrolysis was obtained.
Further, 9.5 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at −5 ° C. for 5 days and then filtered, and 8.6 g of the Nagasaro Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.350 (w / v )%).
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中のウスニン酸の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表１６に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表１６に示す。 Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment. The measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 16. In addition, the theoretical value of the usnic acid content contained in the Nagasaru Ogase extract after the hydrotreatment is also shown in Table 16.
Figure JPOXMLDOC01-appb-T000018
Figure JPOXMLDOC01-appb-T000018
実施例１－５９
　実施例１－５８と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水２．５ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物１２ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１０．５ｇ（蒸発残分０．３００（ｗ／ｖ）％）を得た。 Example 1-59
Digestion was performed by mixing 10 mL of Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 1-58 and 2.5 mL of distilled water, and extraction was performed after Nagasal Ogase after hydrolysis. I got a thing.
Further, 12 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at −5 ° C. for 5 days and filtered, and then 10.5 g of the Nagasaga Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.300 (w / v)%) )
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中のウスニン酸の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表１７に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表１７に示す。 Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment. The measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 17. Table 17 also shows the theoretical value of the usnic acid content contained in the Nagasaru Ogase extract after the hydrotreatment.
Figure JPOXMLDOC01-appb-T000019
Figure JPOXMLDOC01-appb-T000019
実施例１－６０
　実施例１－５８と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水５ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物１４ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１２．６ｇ（蒸発残分０．２４０（ｗ／ｖ）％）を得た。 Example 1-60
Distilled by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 1-58 and 5 mL of distilled water. Obtained.
Furthermore, 14 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 5 days and filtered, and 12.6 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (evaporation residue 0.240 (w / v)%) )
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中のウスニン酸の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表１８に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表１８に示す。 Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment. The measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 18. In addition, the theoretical value of the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment is also shown in Table 18.
Figure JPOXMLDOC01-appb-T000020
Figure JPOXMLDOC01-appb-T000020
実施例１－６１
　実施例１－５８と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水７．５ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物１６ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１４．１ｇ（蒸発残分０．２００（ｗ／ｖ）％）を得た。 Example 1-61
Distillation was performed by mixing 10 mL of Nagasaru Ogase extract before hydrolysis and low-temperature storage prepared in the same manner as in Example 1-58 and 7.5 mL of distilled water, and extraction was performed after Nagasaga Ogase after hydrolysis. I got a thing.
Further, 16 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 5 days and filtered, and 14.1 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.200 (w / v)%) )
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中のウスニン酸の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表１９に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表１９に示す。 Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment. The measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 19. Table 19 also shows the theoretical value of the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment.
Figure JPOXMLDOC01-appb-T000021
Figure JPOXMLDOC01-appb-T000021
実施例１－６２
　実施例１－５８と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水１０ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物のうち１８ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１５．９ｇ（蒸発残分０．１７０（ｗ／ｖ）％）を得た。 Example 1-62
Distilled by mixing 10 mL of Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 1-58 and 10 mL of distilled water, and subjected to hydrolysis, Obtained.
Further, 18 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 5 days and filtered, and then 15.9 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.170 (w / v )%).
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中のウスニン酸の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表２０に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表２０に示す。 Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment. The measurement of the content of usnic acid in the Nagasaru Ogase extract after the hydrotreatment was performed after filtering a part of the extract. The results are shown in Table 20. In addition, the theoretical value of the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment is also shown in Table 20.
Figure JPOXMLDOC01-appb-T000022
Figure JPOXMLDOC01-appb-T000022
　表１６～２０の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理及び低温保存処理をこの順で行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results in Tables 16 to 20, the Nagasaruogase extract obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to a hydrolysis treatment and a low-temperature storage treatment in this order. The usnic acid content contained in the Nagasaru Ogase extract can be further reduced.
実施例１－６３
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１７０．２４ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。 Example 1-63
Yokowa Sarugase (obtained from Hichem Corp.) and 10 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 170.24 g (Evaporation residue 0.348 (w / v)) %).
実施例１－６４
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１５６．７９ｇ（蒸発残分０．４０２（ｗ／ｖ）％）を得た。 Example 1-64
Yokowa Sarugase (obtained from Hichem) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 156.79 g (evaporation residue 0.402 (w / v)) %).
実施例１－６５
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１６２．１８ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。 Example 1-65
Yokowa Sarugase (obtained from Hichem) 10 g and 50 vol% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 162.18 g (Evaporation residue 0.348 (w / v)) %).
実施例１－６６
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１６６．６１ｇ（蒸発残分０．２５０（ｗ／ｖ）％）を得た。 Example 1-66
Yokowa Sarugase (obtained from Hichem Corp.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days followed by filtration. Yokowa Sarugase extract 166.61 g (evaporation residue 0.250) (W / v)%).
実施例１－６７
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１７６．５５ｇ（蒸発残分０．２３０（ｗ／ｖ）％）を得た。 Example 1-67
Yokowa Sarugase (obtained from Hichem) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 176.55 g (evaporation residue 0.230) (W / v)%).
比較例１－９
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して２０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１４６．４５ｇ（蒸発残分０．４０２（ｗ／ｖ）％）を得た。 Comparative Example 1-9
Yokowa Sarugase (obtained from Hichem) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed and extracted for 4 days at 20 ° C., followed by filtration. Yokowa Sarugase extract 144.65 g (evaporation residue 0.402 (w / v)) %).
比較例１－１０
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１４３．９５ｇ（蒸発残分０．２８８（ｗ／ｖ）％）を得た。 Comparative Example 1-10
Yokowa Sarugase (obtained from Hichem) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 143.95 g (Evaporation residue 0.288 (w / v)) %).
ウスニン酸含有量の測定
　上記実施例１－６３～１－６７、並びに比較例１－９及び１－１０で得られた各ヨコワサルオガセ抽出物中のウスニン酸含有量を、前記と同様の方法により測定した。その結果を表２１に示す。 Measurement of usnic acid content The contents of usnic acid in each of the Yokosawa ogase extracts obtained in Examples 1-63 to 1-67 and Comparative Examples 1-9 and 1-10 were measured in the same manner as described above. It was measured. The results are shown in Table 21.
Figure JPOXMLDOC01-appb-T000023
Figure JPOXMLDOC01-appb-T000023
　表２１の結果から明らかなように、本発明の第１の実施態様の方法によれば、ウスニン酸の含有量が十分に低いヨコワサルオガセ抽出物を得ることができる。
　これに対して、アルコール含有率の高すぎる抽出溶媒を用いてヨコワサルオガセを抽出すると、ウスニン酸含有量が上昇した（比較例１－９及び１－１０）。 As is clear from the results in Table 21, according to the method of the first embodiment of the present invention, a Yokosawa ogase extract having a sufficiently low usnic acid content can be obtained.
On the other hand, when Yokosawa ogase was extracted using an extraction solvent having an excessively high alcohol content, the usnic acid content increased (Comparative Examples 1-9 and 1-10).
　以上のように、サルオガセ属地衣類をアルコール含有率が８０体積％以下のアルコール水溶液を用いて抽出することで、ウスニン酸低含有のサルオガセ属地衣類抽出物を製造することができる。ウスニン酸は肝毒性を有するので、本発明の第１の実施態様によれば、安全性に優れ、エンドセリン作用抑制剤等の皮膚外用剤に好適に用いることができるサルオガセ属地衣類を製造することができる。 As described above, a Salusose lichen extract containing a low amount of usnic acid can be produced by extracting the Ligosa lichen using an aqueous alcohol solution having an alcohol content of 80% by volume or less. Since usnic acid has hepatotoxicity, according to the first embodiment of the present invention, it is possible to produce a Sarugose lichen that is excellent in safety and can be suitably used for an external skin preparation such as an endothelin action inhibitor. it can.
実施例１－６８
　実施例１－６４で得られた抽出物（加水・低温保存処理前のヨコワサルオガセ抽出物）８８．５７ｇを－５℃で３日間保存後、ろ過することにより低温保存処理後のヨコワサルオガセ抽出物８１．６８ｇ（蒸発残分０．３９０（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のヨコワサルオガセ抽出物８０ｍＬと蒸留水２０ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で４日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物８６．１２ｇ（蒸発残分０．３２８（ｗ／ｖ）％）を得た。 Example 1-68
88.57 g of the extract obtained from Example 1-64 (Yokosawa Ogase extract before hydrolyzing and low-temperature storage treatment) was stored at −5 ° C. for 3 days and then filtered to obtain Yokosawa Ogase extract after low-temperature storage treatment. 81.68 g (evaporation residue 0.390 (w / v)%) was obtained.
80 mL of the obtained Yokosawa-ogase extract after low-temperature storage treatment and 20 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Yokosawa-ogase extract after the hydrotreatment.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at -5 ° C. for 4 days and then filtered, and 86.12 g (Yield residue 0.328 (w / v)%) of the Yokosawa ogase extract after the hydration / low temperature preservation treatment Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、低温保存処理後のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表２２に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表２２に示す。 The same method as described above for the Yokosawa Ogase extract before hydrolyzed and cryopreserved, the Yokosawa Ogase extract after the cryopreserved treatment, the Yokosawa Ogase extract after the hydrotreated, and the Yokosawa ogase extract after the hydrolyzed and cryopreserved treatment Was used to measure the usnic acid content. The results are shown in Table 22. In addition, Table 22 also shows the theoretical value of the usnic acid content contained in the Yokosawa ogase extract after the hydrotreatment.
Figure JPOXMLDOC01-appb-T000024
Figure JPOXMLDOC01-appb-T000024
実施例１－６９
　実施例１－６４で得られたヨコワサルオガセ抽出物（加水・低温保存処理前のヨコワサルオガセ抽出物）４０ｍＬと蒸留水１０ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で３日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物４１．０３ｇ（蒸発残分０．２８６（ｗ／ｖ）％）を得た。 Example 1-69
40 mL of the Yokosawa Ogase extract obtained in Example 1-64 (Yokosawa Ogase extract before hydrolyzing and low-temperature storage treatment) and 10 mL of distilled water were mixed to dilute the ethanol volume to 40%, followed by hydrolysis. The later Yokosawa Ogase extract was obtained.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at −5 ° C. for 3 days and then filtered, and 41.03 g (Yield residue 0.286 (w / v)%) of the Yokosawa ogase extract after the hydrolyzing and low-temperature preservation treatment Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表２３に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表２３に示す。 About the Yokosawa ogase extract before the water- and low-temperature storage treatment, the Yokosawa ogase extract after the water-treatment, and the Yokowa-saga ogase extract after the water- and low-temperature storage treatment, the usnic acid content was measured by the same method as described above. The results are shown in Table 23. In addition, Table 23 also shows the theoretical value of the usnic acid content contained in the Yokosawa ogase extract after the hydrotreatment.
Figure JPOXMLDOC01-appb-T000025
Figure JPOXMLDOC01-appb-T000025
実施例１－７０
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、加水・低温保存処理前のヨコワサルオガセ抽出物１６２．１８ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。
　得られた加水・低温保存処理前のヨコワサルオガセ抽出物８０ｍＬと蒸留水２０ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で２日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物８６．９４ｇ（蒸発残分０．３２８（ｗ／ｖ）％）を得た。 Example 1-70
Yokowa Sarugase (obtained from Hichem Corp.) and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days, followed by filtration, and 162.18 g of Yokowa Sarugase extract before hydrolyzing and low-temperature storage treatment (evaporation residue 0). .348 (w / v)%).
80 mL of the obtained Yokosawa-ogase extract before hydrolyzing and low-temperature storage treatment and 20 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain the Yokosawa-ogase extract after the hydrotreatment.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at -5 ° C. for 2 days and then filtered, and 86.94 g (Yield residue 0.328 (w / v)%) of the Yokosawa ogase extract after the hydration / low temperature preservation treatment Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表２４に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表２４に示す。 About the Yokosawa ogase extract before the water- and low-temperature storage treatment, the Yokosawa ogase extract after the water-treatment, and the Yokowa-saga ogase extract after the water- and low-temperature storage treatment, the usnic acid content was measured by the same method as described above. The results are shown in Table 24. In addition, the theoretical value of the usnic acid content contained in the Yokosawa ogase extract after the hydrotreatment is also shown in Table 24.
Figure JPOXMLDOC01-appb-T000026
Figure JPOXMLDOC01-appb-T000026
　表２２～２４の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるヨコワサルオガセ抽出物について、加水処理及び低温保存処理を行なうことにより、ヨコワサルオガセ抽出物に含まれるウスニン酸含有量をさらに低減することができる。 As is apparent from the results of Tables 22 to 24, the Yokosawa ogase extract obtained by using an aqueous alcohol solution having an alcohol content of 80% by volume or less as an extraction solvent is subjected to a hydration treatment and a low-temperature storage treatment to thereby obtain a Yokosawa ogase. The usnic acid content contained in the extract can be further reduced.
実施例１－７１
　ナガサルオガセ（新和物産社より入手）１０ｇと５体積％エタノール水溶液１Ｌとを混合して２０℃で２９時間処理した後ろ過し、ろ液９６１．９８ｇ（蒸発残分０．０８０（ｗ／ｖ）％）を分離した。次に、ろ液から溶媒を分離して、ナガサルオガセの残渣を得た。
　前記ナガサルオガセの残渣３２．４８ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ナガサルオガセ抽出物１７８．４４ｇ（蒸発残分０．１７０（ｗ／ｖ）％）を得た。 Example 1-71
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of 5 volume% ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 96.98 g of filtrate (evaporation residue 0.080 (w / v )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue.
32.48 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration, and Nagasaga Ogase extract 178.44 g (evaporation residue 0.170 (w / v)%) )
実施例１－７２
　ナガサルオガセ（新和物産社より入手）１０ｇと１体積％エタノール水溶液１Ｌとを混合して２０℃で２９時間処理した後ろ過し、ろ液９５９．０１ｇ（蒸発残分０．０７６（ｗ／ｖ）％）を分離した。次に、ろ液から溶媒を分離して、ナガサルオガセの残渣を得た。
　前記ナガサルオガセの残渣３５．４７ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ナガサルオガセ抽出物１８０．０３ｇ（蒸発残分０．１５２（ｗ／ｖ）％）を得た。 Example 1-72
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of a 1% by volume ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 959.01 g of filtrate (evaporation residue 0.076 (w / v )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue.
35.47 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days followed by filtration. The Nagasaga Ogase extract 180.03 g (evaporation residue 0.152 (w / v)%) )
実施例１－７３
　ナガサルオガセ（新和物産社より入手）１０ｇと蒸留水１Ｌとを混合して２０℃で２９時間処理した後ろ過し、ろ液９５０．０１ｇ（蒸発残分０．０８０（ｗ／ｖ）％）を分離した。次に、ろ液から溶媒をを分離して、ナガサルオガセの残渣を得た。
　前記ナガサルオガセの残渣３２．７５ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ナガサルオガセ抽出物１７９．０３ｇ（蒸発残分０．１４４（ｗ／ｖ）％）を得た。 Example 1-73
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of distilled water were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 950.01 g of filtrate (evaporation residue 0.080 (w / v)%) Separated. Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue.
32.75 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration. The Nagasaga Ogase extract 179.03 g (evaporation residue 0.144 (w / v)% )
　上記実施例１－７１～１－７３で得られた各ナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでウスニン酸含有量を測定した。その結果を表２５に示す。 For each Nagasagaogase extract obtained in Examples 1-71 to 1-73, the usnic acid content was measured by HPLC in the same manner as described above. The results are shown in Table 25.
Figure JPOXMLDOC01-appb-T000027
Figure JPOXMLDOC01-appb-T000027
　表２５の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を用いて抽出を行う前に、水系溶媒を用いてサルオガセ属地衣類を処理することにより、ウスニン酸含有量が特に低いサルオガセ属地衣類抽出物を得ることができる。 As is apparent from the results in Table 25, the Usunic acid content is particularly improved by treating the Sarcophagus lichen with an aqueous solvent before extraction with an aqueous alcohol solution having an alcohol content of 80% by volume or less. A low Sargogassian lichen extract can be obtained.
実施例１－７４
　実施例１－７１で得られた抽出物（加水・低温保存処理前のナガサルオガセ抽出物）８０ｍＬと蒸留水２０ｍＬとを混合してエタノールの体積が約４０～５０％となるよう希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。さらに、加水処理後のナガサルオガセ抽出物を－５℃で４日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物８４．３３ｇ（蒸発残分０．１４０（ｗ／ｖ）％）を得た。 Example 1-74
80 mL of the extract obtained in Example 1-71 (Nagasaruogase extract before hydrolyzing and low-temperature storage treatment) and 20 mL of distilled water were mixed to dilute the ethanol volume to about 40-50%. The Nagasagaogase extract after the hydration treatment was obtained. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 4 days and filtered, and 84.33 g of the Nagasao Ogase extract after the hydrous and low temperature preservation treatment (evaporation residue 0.140 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりウスニン酸含有量を測定した。その結果を表２６に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるウスニン酸含有量の理論値も併せて表２６に示す。 Usasic acid content was measured by the same method as described above for the Nagasal Ogase extract before the hydrolyzing and low-temperature storage treatment, the Nagasal Ogase extract after the hydrotreating treatment, and the Nagasal Ogase extract after the hydrolyzing and low-temperature preservation treatment. The results are shown in Table 26. In addition, the theoretical value of the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment is also shown in Table 26.
Figure JPOXMLDOC01-appb-T000028
Figure JPOXMLDOC01-appb-T000028
　表２６の結果から明らかなように、アルコール含有率が８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理及び低温保存処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量を選択的に低減することができる。 As is apparent from the results of Table 26, Nagasaga Ogase extract is obtained by subjecting Nagasaga Ogase extract obtained by using an alcohol aqueous solution having an alcohol content of 80% by volume or less as an extraction solvent to hydrotreating and low-temperature storage treatment. The usnic acid content contained in can be selectively reduced.
（第２の実施態様に関する実施例及び比較例）
　以下、第２の実施態様に関する実施例及び比較例について説明する。 (Examples and comparative examples relating to the second embodiment)
Hereinafter, examples and comparative examples related to the second embodiment will be described.
比較例２－１
　ナガサルオガセ（新和物産社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０７．３５ｇ（蒸発残分０．２０２（ｗ／ｖ）％）を得た。 Comparative Example 2-1
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, 207.35 g of Nagasaru Ogase extract (evaporation residue 0.202 (w / w v)%).
比較例２－２
　ナガサルオガセ（新和物産社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０６．５９ｇ（蒸発残分０．２６６（ｗ／ｖ）％）を得た。 Comparative Example 2-2
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed and extracted at 30 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 206.59 g (evaporation residue 0.266 (w / v)%).
比較例２－３
　ナガサルオガセ（新和物産社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０８．２６ｇ（蒸発残分０．２８０（ｗ／ｖ）％）を得た。 Comparative Example 2-3
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 10 volume% ethanol aqueous solution 200 mL were mixed, extracted at 40 ° C. for 7 days, filtered, and Nagasal Ogase extract 208.26 g (evaporation residue 0.280 (w / w v)%).
比較例２－４
　ナガサルオガセ（新和物産社より入手）１０ｇと２０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０１．４９ｇ（蒸発残分０．１７２（ｗ／ｖ）％）を得た。 Comparative Example 2-4
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 20 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.49 g (evaporation residue 0.172 (w / v)%).
実施例２－１
　ナガサルオガセ（新和物産社より入手）１０ｇと３０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．９０ｇ（蒸発残分０．１４６（ｗ／ｖ）％）を得た。 Example 2-1
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 30 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 199.90 g (evaporation residue 0.146 (w / v)%).
実施例２－２
　ナガサルオガセ（新和物産社より入手）１０ｇと４０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２００．７０ｇ（蒸発残分０．１９０（ｗ／ｖ）％）を得た。 Example 2-2
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 40 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasal Ogase extract 200.70 g (evaporation residue 0.190 (w / v)%).
実施例２－３
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０１．６７ｇ（蒸発残分０．１７８（ｗ／ｖ）％）を得た。 Example 2-3
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 5 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.67 g (evaporation residue 0.178 (w / v)%).
実施例２－４
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して１５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０２．５７ｇ（蒸発残分０．１９２（ｗ／ｖ）％）を得た。 Example 2-4
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 15 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 202.57 g (evaporation residue 0.192 (w / v)%).
実施例２－５
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．６９ｇ（蒸発残分０．２９８（ｗ／ｖ）％）を得た。 Example 2-5
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.69 g (evaporation residue 0.298 (w / v)%).
実施例２－６
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２００．５４ｇ（蒸発残分０．２５８（ｗ／ｖ）％）を得た。 Example 2-6
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 30 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 200.54 g (evaporation residue 0.258 (w / v)%).
実施例２－７
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．４９ｇ（蒸発残分０．３１６（ｗ／ｖ）％）を得た。 Example 2-7
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% aqueous ethanol solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration, Nagasaru Ogase extract 199.49 g (evaporation residue 0.316 (w / v)%).
実施例２－８
　ナガサルオガセ（新和物産社より入手）１０ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９８．６９ｇ（蒸発残分０．２４２（ｗ／ｖ）％）を得た。 Example 2-8
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 60 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 198.69 g (evaporation residue 0.242 (w / v)%).
実施例２－９
　ナガサルオガセ（新和物産社より入手）１０ｇと７０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９５．１ｇ（蒸発残分０．２３２（ｗ／ｖ）％）を得た。 Example 2-9
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 70 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, 195.1 g of Nagasal Ogase extract (evaporation residue 0.232 (w / v)%).
実施例２－１０
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８５．６ｇ（蒸発残分０．２１６（ｗ／ｖ）％）を得た。 Example 2-10
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 80 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.6 g (evaporation residue 0.216 (w / v)%).
比較例２－５
　ナガサルオガセ（新和物産社より入手）１０ｇと９０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８３．４ｇ（蒸発残分０．１９２（ｗ／ｖ）％）を得た。 Comparative Example 2-5
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 90 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaga Ogase extract 183.4 g (evaporation residue 0.192 (w / v)%).
比較例２－６
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８４．２４ｇ（蒸発残分０．１２８（ｗ／ｖ）％）を得た。 Comparative Example 2-6
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 5 ° C. for 7 days, filtered, and Nagasaru Ogase extract 184.24 g (evaporation residue 0.128 (w / v)%).
比較例２－７
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して１５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８４．０ｇ（蒸発残分０．１５４（ｗ／ｖ）％）を得た。 Comparative Example 2-7
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% aqueous ethanol solution 200 mL were mixed and extracted at 15 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 184.0 g (evaporation residue 0.154 (w / v)%).
比較例２－８
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８４．２２ｇ（蒸発残分０．１７４（ｗ／ｖ）％）を得た。 Comparative Example 2-8
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% aqueous ethanol solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 184.22 g (evaporation residue 0.174 (w / v)%).
比較例２－９
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８６．４２ｇ（蒸発残分０．２１８（ｗ／ｖ）％）を得た。 Comparative Example 2-9
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 30 ° C. for 7 days, filtered, 186.42 g of Nagasal Ogase extract (evaporation residue 0.218 (w / v)%).
比較例２－１０
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８５．０６ｇ（蒸発残分０．２６６（ｗ／ｖ）％）を得た。 Comparative Example 2-10
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 40 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.06 g (evaporation residue 0.266 (w / v)%).
比較例２－１１
　ナガサルオガセ（新和物産社より入手）２５ｇと９９．５体積％エタノール水溶液５００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物３６８．０ｇ（蒸発残分０．１５８（ｗ／ｖ）％）を得た。 Comparative Example 2-11
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 25 g and 99.5 vol% ethanol aqueous solution 500 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 368.0 g (evaporation residue 0.158 ( w / v)%).
ジフラクタ酸含有量及びウスニン酸含有量の測定
　上記実施例２－１～２－１０及び比較例２－１～２－１１で得られた各ナガサルオガセ抽出物中のジフラクタ酸含有量及びウスニン酸含有量を、ＨＰＬＣを用いて下記の条件で分析した。測定に用いた装置及び測定条件は下記のとおりである。
 
測定機器(HPLC)：Agilent　Technologies　Open　LAB　CDS　LC02-1260、オートサンプラーG1329B　1260　ALS、カラムオーブンG1316A　1260　TCC、検出器G1315D　1260　DAD　VL、ポンプG1312B　1260　Bin　pump、G1379B　1260　μ-Degasser
 
＜HPLC分析条件＞
流速：０．７５ｍＬ/ｍｉｎ
使用したカラム：Inertsil　ODS-3　５μｍ(GL　Science)、3.0I.D.×１５０ｍｍ
検出波長：２５４ｎｍ
カラム温度：４０℃
移動相：
Ａ：0.1%トリフルオロ酢酸含有水溶液、Ｂ：0.1%トリフルオロ酢酸含有メタノール
　　　０～１０分：70%Ｂ
　　１０～１５分：70％Ｂ→100%Ｂ
　　１５～２０分：100％Ｂ
 
　結果を表２７に示す。 Measurement of difractic acid content and usnic acid content Difractic acid content and usnic acid content in each Nagasagaogase extract obtained in Examples 2-1 to 2-10 and Comparative Examples 2-1 to 2-11 The amount was analyzed using HPLC under the following conditions. The apparatus and measurement conditions used for the measurement are as follows.

Measuring instruments (HPLC): Agilent Technologies Open LAB CDS LC02-1260, Autosampler G1329B 1260 ALS, Column oven G1316A 1260 TCC, Detector G1315D 1260 DAD VL, Pump G1312B 1260 Bin pump, G1379B 1260 μ-Degasser

<HPLC analysis conditions>
Flow rate: 0.75 mL / min
Column used: Inertsil ODS-3 5 μm (GL Science), 3.0 ID × 150 mm
Detection wavelength: 254 nm
Column temperature: 40 ° C
Mobile phase:
A: 0.1% trifluoroacetic acid-containing aqueous solution, B: 0.1% trifluoroacetic acid-containing methanol 0 to 10 minutes: 70% B
10-15 minutes: 70% B → 100% B
15-20 minutes: 100% B

The results are shown in Table 27.
Figure JPOXMLDOC01-appb-T000029
Figure JPOXMLDOC01-appb-T000029
　表２７の結果から明らかなように、本発明の第２の実施態様の方法によれば、ジフラクタ酸含有量が十分に高くウスニン酸含有量が低減した、ジフラクタ酸高含有かつウスニン酸低含有のナガサルオガセ抽出物を得ることができる。これに対して、アルコール含有率の低すぎる抽出溶媒を用いてナガサルオガセを抽出すると、ウスニン酸含有量は低下したが、ジフラクタ酸含有量も大きく低下した（比較例２－１～２－４）。また、アルコール含有率の高すぎる抽出溶媒を用いてナガサルオガセを抽出すると、ジフラクタ酸含有量は上昇するが、ウスニン酸含有量も上昇した（比較例２－５～２－１１）。
　以上のように、サルオガセ属地衣類をアルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出することで、ジフラクタ酸高含有かつウスニン酸低含有のサルオガセ属地衣類抽出物を製造することができる。ウスニン酸は肝毒性を有する一方、ジフラクタ酸は優れたエンドセリン抑制作用を有する。したがって、本発明の第２の実施態様によれば、エンドセリン作用抑制効果を有し安全性に優れたエンドセリン作用抑制剤を製造することができる。 As is clear from the results in Table 27, according to the method of the second embodiment of the present invention, the difractic acid content was sufficiently high and the usnic acid content was reduced, and the difractic acid high content and the usnic acid low content were low. Nagasarogase extract can be obtained. On the other hand, when Nagasagaogase was extracted using an extraction solvent having an alcohol content that was too low, the usnic acid content decreased, but the difractic acid content also significantly decreased (Comparative Examples 2-1 to 2-4). . In addition, when Nagasagaogase was extracted using an extraction solvent having an alcohol content that was too high, the content of difractate increased, but the content of usnic acid also increased (Comparative Examples 2-5 to 2-11).
As described above, an extract of Salgogase lichen having a high content of difractate and a low content of usnic acid is produced by extracting the Licorice lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume. be able to. Usnic acid has hepatotoxicity, while difractic acid has an excellent endothelin inhibitory action. Therefore, according to the second embodiment of the present invention, an endothelin action inhibitor having an endothelin action inhibitory effect and excellent in safety can be produced.
実施例２－１１
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で８日間抽出後ろ過し、ナガサルオガセ抽出物８５８．５ｇ（蒸発残分０．３７４（ｗ／ｖ）％）を得た。
　得られた抽出物を－５℃で１２日間保存後ろ過し、低温保存処理後のナガサルオガセ抽出物８３３．４ｇ（蒸発残分０．３６０（ｗ／ｖ）％）を得た。 Example 2-11
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasaga Ogase extract 858.5 g (evaporation residue 0.374 (w / v)%).
The obtained extract was stored at −5 ° C. for 12 days and then filtered to obtain 833.4 g of Nagasaroogase extract (low evaporation residue 0.360 (w / v)%) after the low-temperature storage treatment.
　低温保存処理前後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表２８に示す。 For the Nagasaruogase extract before and after the low-temperature storage treatment, the difractic acid content and the usnic acid content were measured by the same method as described above. The results are shown in Table 28.
Figure JPOXMLDOC01-appb-T000030
Figure JPOXMLDOC01-appb-T000030
　表２８の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を保持したままウスニン酸含有量を選択的に低減することができる。 As is clear from the results in Table 28, Nagasagaogase extract is obtained by subjecting Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent to a low-temperature storage treatment. The usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the product.
実施例２－１２
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で８日間抽出後ろ過し、ナガサルオガセ抽出物８５２．８ｇ（蒸発残分０．２６８（ｗ／ｖ）％）を得た。得られた抽出物を－５℃で１３日間保存後、ろ過することにより、ナガサルオガセ抽出物処理前品８３７．２ｇ（蒸発残分０．２５６（ｗ／ｖ）％）を得た。
　次に、ナガサルオガセ抽出物処理前品１０ｍＬと蒸留水２．５ｍＬとを混合してエタノールの体積が４０％となるよう希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。さらに、加水処理後のナガサルオガセ抽出物を－５℃で４日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１１．１ｇ（蒸発残分０．１３０（ｗ／ｖ）％）を得た。 Example 2-12
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed, extracted at 20 ° C. for 8 days, filtered, and Nagasal Ogase extract 852.8 g (evaporation residue 0.268 (w / w v)%). The obtained extract was stored at −5 ° C. for 13 days and filtered to obtain 837.2 g (evaporation residue 0.256 (w / v)%) of the Nagasaga Ogase extract before treatment.
Next, 10 mL of Nagasaga Ogase extract pretreatment product and 2.5 mL of distilled water were mixed and diluted so that the volume of ethanol was 40%. . Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 4 days and then filtered, and the Nagasal Ogase extract after the hydrous / low temperature preservation treatment was 11.1 g (evaporation residue 0.130 (w / v)%). Got.
　ナガサルオガセ抽出物処理前品、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表２９に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表２９に示す。 The pretreatment with Nagasaru Ogase extract, the Nagasal Ogase extract after the hydrotreatment, and the Nagasal Ogase extract after the hydrothermal treatment at low temperature were measured for difractic acid content and usnic acid content by the same method as described above. The results are shown in Table 29. Table 29 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment.
Figure JPOXMLDOC01-appb-T000031
Figure JPOXMLDOC01-appb-T000031
　表２９の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理、加水処理及び低温保存処理をこの順で行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量を選択的に低減することができる。 As is clear from the results in Table 29, Nagasagaogase extract obtained using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to low-temperature storage treatment, hydrolysis treatment and low-temperature storage treatment. By carrying out in this order, the usnic acid content contained in the Nagasaga Ogase extract can be selectively reduced.
実施例２－１３
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物８３８．５ｇ（蒸発残分０．２１８（ｗ／ｖ）％）を得た。
　次に、加水・低温保存処理前のナガサルオガセ抽出物８００ｍＬと蒸留水２００ｍＬとを混合してエタノールの体積が４０％となるよう希釈し澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。さらに、加水処理後のナガサルオガセ抽出物を－５℃で７日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物９１６．２ｇ（蒸発残分０．１３０（ｗ／ｖ）％）を得た。 Example 2-13
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 1 L of 50% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 7 days, filtered, and 838.5 g (Evaporation residue) Min 0.218 (w / v)%).
Next, 800 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 200 mL of distilled water are mixed and diluted so that the volume of ethanol becomes 40%, and then the Nagasaga Ogase extract after hydrolysis is obtained. It was. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 7 days and filtered, and 916.2 g of the Nagasal Ogase extract after the hydrothermal treatment at low temperature (Evaporation residue 0.130 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表６に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表３０に示す。 For the Nagasaru Ogase extract before the water-and-low-temperature storage treatment and for the Nagasal-ogase extract after the water-and-low-temperature storage treatment, the difractic acid content and the usnic acid content were measured by the same method as described above. The results are shown in Table 6. In addition, the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment are also shown in Table 30.
Figure JPOXMLDOC01-appb-T000032
Figure JPOXMLDOC01-appb-T000032
　表３０の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理及び低温保存処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を保持したままウスニン酸含有量を選択的に低減することができる。 As is apparent from the results in Table 30, the Nagasaruogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a hydration treatment and a low-temperature storage treatment. The usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the Nagasaru Ogase extract.
実施例２－１４
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）０．６５ｍｇ（抽出物中固形分に対して１０重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭１０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．０８（ｗ／ｖ）％））。その結果を表３１に示す。 Example 2-14
5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. Then, add 0.65 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (10% by weight based on the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 10 wt% -treated product was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.08 (w / v)%)). The results are shown in Table 31.
実施例２－１５
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）３．２５ｍｇ（抽出物中固形分に対して５０重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表３１に示す。 Example 2-15
5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. After dilution, add 3.25 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (50% by weight based on the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 50 wt% treated product was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
実施例２－１６
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）６．５０ｍｇ（抽出物中固形分に対して１００重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭１００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表３１に示す。 Example 2-16
5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. After dilution, add 6.50 mg of activated carbon (trade name: Shirahige M, manufactured by Nippon Enviro Chemicals) (100% by weight with respect to the solid content in the extract), stir at room temperature for 45 minutes, and then filter. As a result, an activated carbon 100 wt% treated product was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
実施例２－１７
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）９．７５ｍｇ（抽出物中固形分に対して１５０重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭１５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表３１に示す。 Example 2-17
5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. Then, 9.75 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (150% by weight with respect to the solid content in the extract) is added, and the mixture is stirred at room temperature for 45 minutes and then filtered. Thus, a product after treatment with 150 wt% of activated carbon was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
実施例２－１８
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物（活性炭処理前のナガサルオガセ抽出物）５ｍＬ（固形分６．５ｍｇ）と９９．５体積％エタノール水溶液４５ｍＬとを混合して希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１３．０ｍｇ（抽出物中固形分に対して２００重量％）を添加し、室温で４５分間撹拌後、ろ過することにより活性炭２００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で５ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．０７（ｗ／ｖ）％））。その結果を表３１に示す。 Example 2-18
5 mL of Nagasaru Ogase extract (Nagasal Ogase extract before activated carbon treatment) obtained in Example 2-13 (solid content 6.5 mg) and 45 mL of 99.5 vol% ethanol aqueous solution were mixed. 13.0 mg of activated carbon (trade name: Shirasagi M, manufactured by Nippon Enviro Chemicals) (200% by weight with respect to the solid content in the extract) is added, stirred for 45 minutes at room temperature, and then filtered. Thus, a product after treatment with 200 wt% of activated carbon was obtained. The obtained activated carbon-treated product was dissolved in 5 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.07 (w / v)%)). The results are shown in Table 31.
Figure JPOXMLDOC01-appb-T000033
Figure JPOXMLDOC01-appb-T000033
　表３１の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理、低温保存処理及び活性炭処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を保持したままウスニン酸含有量を選択的に低減することができる。 As is clear from the results in Table 31, the Nagasagaogase extract obtained using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and activated carbon treatment. Thus, the usnic acid content can be selectively reduced while maintaining the content of difractacic acid contained in the Nagasaroogase extract.
実施例２－１９
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物（ヘキサン洗浄処理前抽出物）５０ｍＬにヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄１回品）、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した。その結果を表３２に示す。 Example 2-19
50 mL of hexane and 5 mL of 99.5 vol% ethanol aqueous solution were added to 50 mL of Nagasaroogase extract (extract before hexane washing treatment) obtained in Example 2-13 after hydrolyzing and low-temperature storage treatment, and a separatory funnel was used. After separating into two layers, the lower layer was recovered (hexane washed once product), and the contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 32.
実施例２－２０
　実施例２－１９で得られたナガサルオガセ抽出物５０ｍＬに、再度ヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄２回品）、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した。その結果を表３２に示す。 Example 2-20
To 50 mL of Nagasaga Ogase extract obtained in Example 2-19, 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution were added again, and after separating into two layers using a separatory funnel, the lower layer was recovered (hexane The contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 32.
実施例２－２１
　実施例２－２０で得られたナガサルオガセ抽出物４５ｍＬに、再度ヘキサン４５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄３回品）、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した。その結果を表３２に示す。 Example 2-21
Add 45 mL of hexane again to 45 mL of Nagasaga Ogase extract obtained in Example 2-20, separate into two layers using a separatory funnel, and recover the lower layer (hexane washed three times), as above The contents of difractic acid and usnic acid were measured by HPLC according to the above method. The results are shown in Table 32.
Figure JPOXMLDOC01-appb-T000034
Figure JPOXMLDOC01-appb-T000034
　表３２の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理、低温保存処理及びヘキサン洗浄処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を保持したままウスニン酸含有量を選択的に低減することができる。 As is apparent from the results in Table 32, Nagasagaogase extract obtained by using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to hydration treatment, low-temperature storage treatment, and hexane washing treatment. By carrying out, it is possible to selectively reduce the usnic acid content while maintaining the content of difractic acid contained in the Nagasaroogase extract.
実施例２－２２
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で３日間抽出後ろ過し、ナガサルオガセ抽出物８５８．３ｇ（蒸発残分０．４６０（ｗ／ｖ）％）を得た。
　得られたナガサルオガセ抽出物を－５℃で１２日間保存後、ろ過することにより活性炭処理前のナガサルオガセ抽出物８４９．２ｇ（蒸発残分０．３６０（ｗ／ｖ）％）を得た。
　得られた活性炭処理前のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表３３に示す。 Example 2-22
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% aqueous ethanol solution 1 L were mixed and extracted at 20 ° C. for 3 days followed by filtration. Nagasaru Ogase extract 858.3 g (evaporation residue 0.460 (w / v)%).
The obtained Nagasaga Ogase extract was stored at −5 ° C. for 12 days and then filtered to obtain 849.2 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before activated carbon treatment.
About the obtained Nagasaruogase extract before the activated carbon treatment, the difractic acid content and the usnic acid content were measured by the same method as described above. The results are shown in Table 33.
実施例２－２３
　実施例２－２２で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１．０８ｍｇ（抽出物中固形分に対して１０重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭１０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．２８０（ｗ／ｖ）％））。その結果を表３３に示す。 Example 2-23
27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) 1.08 mg (manufactured by Enviro Chemicals) (10% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with 10 wt% of activated carbon. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 33.
実施例２－２４
　実施例２－２２で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）５．４０ｍｇ（抽出物中固形分に対して５０重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．２８０（ｗ／ｖ）％））。その結果を表３３に示す。 Example 2-24
27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 5.40 mg (50 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon 50 wt%. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.280 (w / v)%)). The results are shown in Table 33.
実施例２－２５
　実施例２－２２で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１０．８０ｍｇ（抽出物中固形分に対して１００重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭１００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．２７０（ｗ／ｖ）％））。その結果を表３３に示す。 Example 2-25
27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 10.80 mg (100% by weight based on solid content in the extract) was added, stirred for 30 minutes at room temperature, and then filtered to obtain a product after treatment with activated carbon 100 wt%. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 33.
実施例２－２６
　実施例２－２２で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）１６．２０ｍｇ（抽出物中固形分に対して１５０重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭１５０ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．２７０（ｗ／ｖ）％））。その結果を表３３に示す。 Example 2-26
27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd.) 16.20 mg (150 wt% with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with 150 wt% of activated carbon. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 33.
実施例２－２７
　実施例２－２２で得られた活性炭処理前のナガサルオガセ抽出物３ｍＬ（固形分１０．８ｍｇ）に９９．５体積％エタノール水溶液２７ｍＬを加え、希釈した後、活性炭（商品名：白鷺Ｍ、日本エンバイロケミカルズ社製）２１．６０ｍｇ（抽出物中固形分に対して２００重量％）を添加し、室温で３０分間撹拌後、ろ過することにより活性炭２００ｗｔ％処理後品を得た。得られた活性炭処理品は活性炭処理時の希釈前品と同容量とすべく減圧下で溶媒を留去した後で３ｍＬの５０体積％エタノール水溶液に溶解し、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．２７０（ｗ／ｖ）％））。その結果を表３３に示す。 Example 2-27
27 mL of a 99.5% by volume ethanol aqueous solution was added to 3 mL of Nagasaroogase extract (solid content 10.8 mg) before activated carbon treatment obtained in Example 2-22, diluted, and then activated carbon (trade name: Shirasagi M, Japan) Enviro Chemicals Co., Ltd. (21.60 mg) (200% by weight with respect to the solid content in the extract) was added, stirred for 30 minutes at room temperature, and filtered to obtain a product after treatment with activated carbon of 200 wt%. The obtained activated carbon-treated product was dissolved in 3 mL of 50% by volume ethanol aqueous solution after distilling off the solvent under reduced pressure so as to have the same volume as the product before dilution at the time of the activated carbon treatment. The acid and usnic acid contents were measured (evaporation residue 0.270 (w / v)%)). The results are shown in Table 33.
Figure JPOXMLDOC01-appb-T000035
Figure JPOXMLDOC01-appb-T000035
　表３３の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理及び活性炭処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を大幅にロスすることなく、ウスニン酸含有量を選択的に低減することができる。 As apparent from the results in Table 33, by performing low-temperature storage treatment and activated carbon treatment on the Nagasaruogase extract obtained using an alcohol aqueous solution having an alcohol content of 30% by volume to 80% by volume as an extraction solvent, The usnic acid content can be selectively reduced without significantly losing the content of difractic acid contained in the Nagasaru Ogase extract.
実施例２－２８
　ナガサルオガセ（新和物産社より入手）５０ｇと５０体積％エタノール水溶液１Ｌとを混合して２０℃で８日間抽出後ろ過し、ナガサルオガセ抽出物８５５．８ｇ（蒸発残分０．２７０（ｗ／ｖ）％）を得た。
　得られたナガサルオガセ抽出物を－５℃で１３日間保存後、ろ過することによりヘキサン洗浄処理前のナガサルオガセ抽出物８３９．５ｇ（蒸発残分０．３６０（ｗ／ｖ）％）を得た。
　得られたヘキサン洗浄処理前のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表３４に示す。 Example 2-28
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 50 L% ethanol aqueous solution 1 L were mixed and extracted at 20 ° C. for 8 days followed by filtration. Nagasaru Ogase extract 855.8 g (evaporation residue 0.270 (w / v)%).
The obtained Nagasaga Ogase extract was stored at −5 ° C. for 13 days and then filtered to obtain 839.5 g (Evaporation residue 0.360 (w / v)%) of Nagasaga Ogase extract before hexane washing treatment. .
About the Nagasaruogase extract before the obtained hexane washing process, the difractic acid content and the usnic acid content were measured by the method similar to the above. The results are shown in Table 34.
実施例２－２９
　実施例２－２８で得られたヘキサン洗浄処理前のナガサルオガセ抽出物５０ｍＬにヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄１回品）、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した。その結果を表３４に示す。 Example 2-29
Add 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution to 50 mL of Nagasaga Ogase extract obtained in Example 2-28 before washing with hexane, separate into two layers using a separatory funnel, and recover the lower layer Then, the contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 34.
実施例２－３０
　実施例２－２９で得られたナガサルオガセ抽出物５０ｍＬに、再度ヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄２回品）、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した。その結果を表３４に示す。 Example 2-30
To 50 mL of Nagasaga Ogase extract obtained in Example 2-29, 50 mL of hexane and 5 mL of a 99.5% by volume ethanol aqueous solution were added again. After separating into two layers using a separatory funnel, the lower layer was recovered (hexane The contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 34.
実施例２－３１
　実施例２－３０で得られたナガサルオガセ抽出物５０ｍＬに、再度ヘキサン５０ｍＬ及び９９．５体積％エタノール水溶液５ｍＬを加え、分液ロートを使用して２層に分離後、下層を回収し（ヘキサン洗浄３回品）、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した（蒸発残分０．２８６（ｗ／ｖ）％）。その結果を表３４に示す。 Example 2-31
To 50 mL of Nagasaga Ogase extract obtained in Example 2-30, 50 mL of hexane and 5 mL of 99.5% by volume ethanol aqueous solution were added again, and after separating into two layers using a separatory funnel, the lower layer was recovered (hexane The contents of difractic acid and usnic acid were measured by HPLC by the same method as described above (evaporation residue 0.286 (w / v)%). The results are shown in Table 34.
Figure JPOXMLDOC01-appb-T000036
Figure JPOXMLDOC01-appb-T000036
　表３４の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理及びヘキサン洗浄処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を大幅にロスすることなく、ウスニン酸含有量を選択的に低減することができる。 As is apparent from the results in Table 34, Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a low-temperature storage treatment and a hexane washing treatment. The usnic acid content can be selectively reduced without significantly losing the content of difractic acid contained in the Nagasaru Ogase extract.
実施例２－３２～２－３９
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物１０ｍＬを６０℃で静置保存し、表３５に示した各保存時間後のナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでジフラクタ酸とウスニン酸の含有量を測定した。その結果を表３５に示す。 Examples 2-32 to 2-39
10 mL of the Nagasaga Ogase extract after hydrolyzed and cryopreserved treatment obtained in Example 2-13 was stored at 60 ° C., and the Nagasaga Ogase extract after each storage time shown in Table 35 was the same as described above. The contents of difractic acid and usnic acid were measured by HPLC according to the method. The results are shown in Table 35.
Figure JPOXMLDOC01-appb-T000037
Figure JPOXMLDOC01-appb-T000037
実施例２－４０～２－４３
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物１５０ｍＬを７０℃で表１２に示した時間で攪拌を行い、得られたナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでジフラクタ酸とウスニン酸の含有量を測定した。その結果を表３６に示す。 Examples 2-40 to 2-43
150 mL of the Nagasaga Ogase extract obtained after the hydrolyzing and low-temperature storage treatment obtained in Example 2-13 was stirred at 70 ° C. for the time shown in Table 12, and the obtained Nagasaga Ogase extract was subjected to the same method as described above. Was used to measure the contents of difractic acid and usnic acid by HPLC. The results are shown in Table 36.
Figure JPOXMLDOC01-appb-T000038
Figure JPOXMLDOC01-appb-T000038
実施例２－４４～２－４６
　実施例２－１３で得られた加水・低温保存処理後のナガサルオガセ抽出物１５０ｍＬを８０℃で表１３に示した時間で攪拌を行い、得られたナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでジフラクタ酸とウスニン酸の含有量を測定した。その結果を表３７に示す。 Examples 2-44 to 2-46
150 mL of the Nagasaga Ogase extract obtained after the hydrolyzing and low-temperature storage treatment obtained in Example 2-13 was stirred at 80 ° C. for the time shown in Table 13, and the obtained Nagasaga Ogase extract was subjected to the same method as described above. Was used to measure the contents of difractic acid and usnic acid by HPLC. The results are shown in Table 37.
Figure JPOXMLDOC01-appb-T000039
Figure JPOXMLDOC01-appb-T000039
　表３５～３７の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理、低温保存処理及び加熱処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を保持したままウスニン酸含有量を選択的に低減することができる。 As is clear from the results in Tables 35 to 37, the Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to water treatment, low-temperature storage treatment, and heat treatment. By carrying out the process, the usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the Nagasaruogase extract.
実施例２－４７
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７５．６２ｇ（蒸発残分０．２９０（ｗ／ｖ）％）を得た。 Example 2-47
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaga Ogase extract 175.62 g (evaporation residue 0 .290 (w / v)%).
実施例２－４８
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７４．２５ｇ（蒸発残分０．３４０（ｗ／ｖ）％）を得た。 Example 2-48
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and 174.25 g of Nagasal Ogase extract (evaporation residue 0 .340 (w / v)%).
実施例２－４９
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７７．５７ｇ（蒸発残分０．４１０（ｗ／ｖ）％）を得た。 Example 2-49
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 177.57 g (evaporation residue 0 .410 (w / v)%).
実施例２－５０
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７８．１９ｇ（蒸発残分０．２６０（ｗ／ｖ）％）を得た。 Example 2-50
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaga Ogase extract 178.19 g (evaporation residue 0 260 (w / v)%).
実施例２－５１
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７９．８２ｇ（蒸発残分０．３００（ｗ／ｖ）％）を得た。 Example 2-51
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed, extracted at 30 ° C. for 7 days, filtered, and Nagasal Ogase extract 179.82 g (evaporation residue 0 .300 (w / v)%).
実施例２－５２
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８２．１４ｇ（蒸発残分０．３８０（ｗ／ｖ）％）を得た。 Example 2-52
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 182.14 g (evaporation residue 0 .380 (w / v)%).
比較例２－１２
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２５℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８２．５５ｇ（蒸発残分０．１２（ｗ／ｖ）％）を得た。 Comparative Example 2-12
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% 1,3-butanediol aqueous solution 200 mL were mixed, extracted at 25 ° C. for 7 days, filtered, and Nagasaga Ogase extract 182.55 g (evaporation residue 0 .12 (w / v)%).
ジフラクタ酸含有量及びウスニン酸含有量の測定
　上記実施例２－４７～２－５２及び比較例２－１２で得られた各ナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した。その結果を表３８に示す。 Measurement of Difractic Acid Content and Usnic Acid Content For each of Nagasaruogase extracts obtained in Examples 2-47 to 2-52 and Comparative Example 2-12, difractic acid and usnin were extracted by HPLC in the same manner as described above. The acid content was measured. The results are shown in Table 38.
Figure JPOXMLDOC01-appb-T000040
Figure JPOXMLDOC01-appb-T000040
　表３８の結果から明らかなように、本発明の第２の実施態様の方法によれば、ジフラクタ酸含有量が十分に高くウスニン酸含有量が低減した、ジフラクタ酸高含有かつウスニン酸低含有のナガサルオガセ抽出物を得ることができる。これに対して、アルコール含有率の高すぎる抽出溶媒を用いてナガサルオガセを抽出すると、ジフラクタ酸濃度は上昇するが、ウスニン酸濃度も上昇した（比較例２－１２）。
　以上のように、サルオガセ属地衣類をアルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出することで、ジフラクタ酸高含有かつウスニン酸低含有のサルオガセ属地衣類の抽出物を製造することができる。ウスニン酸は肝毒性を有する一方、ジフラクタ酸は優れたエンドセリン抑制作用を有する。したがって、本発明の第２の実施態様によれば、エンドセリン作用抑制効果を有し安全性に優れたエンドセリン作用抑制剤を製造することができる。 As is apparent from the results of Table 38, according to the method of the second embodiment of the present invention, the difractic acid content was sufficiently high and the usnic acid content was reduced, and the difractic acid high content and the low usnic acid content were low. Nagasarogase extract can be obtained. On the other hand, when Nagasaruogase was extracted using an extraction solvent with an excessively high alcohol content, the concentration of difractate increased, but the concentration of usnic acid also increased (Comparative Example 2-12).
As described above, the extract of Salogase lichen having a high content of difractic acid and a low content of usnic acid is produced by extracting the Licorice lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume. can do. Usnic acid has hepatotoxicity, while difractic acid has an excellent endothelin inhibitory action. Therefore, according to the 2nd embodiment of this invention, the endothelin action inhibitor which has an endothelin action inhibitory effect and was excellent in safety | security can be manufactured.
実施例２－５３
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１７７．６ｇ（蒸発残分０．４２０（ｗ／ｖ）％）を得た。
　得られた抽出物１０ｍＬを－５℃で５日間保存後ろ過し、低温保存処理後のナガサルオガセ抽出物８．９ｇ（蒸発残分０．３８０（ｗ／ｖ）％）を得た。 Example 2-53
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract (177.6 g) 420 (w / v)%).
10 mL of the obtained extract was stored at −5 ° C. for 5 days and then filtered to obtain 8.9 g of Nagasaga Ogase extract after evaporation at low temperature (evaporation residue 0.380 (w / v)%).
　低温保存処理前後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表３９に示す。 For the Nagasaruogase extract before and after the low-temperature storage treatment, the difractic acid content and the usnic acid content were measured by the same method as described above. The results are shown in Table 39.
Figure JPOXMLDOC01-appb-T000041
Figure JPOXMLDOC01-appb-T000041
　表３９の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、低温保存処理を行なうことにより、ナガサルオガセ抽出物に含まれるジフラクタ酸の含有量を保持したままウスニン酸含有量を選択的に低減することができる。 As is clear from the results in Table 39, Nagasaga Ogase extract is obtained by subjecting Nagasaga Ogase extract obtained by using an alcohol aqueous solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent to a low-temperature storage treatment. The usnic acid content can be selectively reduced while maintaining the content of difractic acid contained in the product.
実施例２－５４
　ナガサルオガセ（新和物産社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物１７７．６ｇ（蒸発残分０．４２０（ｗ／ｖ）％）を得た。
　次に、加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水１ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物９．５ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物８．６ｇ（蒸発残分０．３５０（ｗ／ｖ）％）を得た。 Example 2-54
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 50 volume% 1,3-butanediol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 7 days, followed by filtration. 0.6 g (evaporation residue 0.420 (w / v)%) was obtained.
Next, the mixture was diluted by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 1 mL of distilled water to perform starch, and a Nagasal Ogase extract after hydrolysis was obtained.
Further, 9.5 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at −5 ° C. for 5 days and then filtered, and 8.6 g of the Nagasaro Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.350 (w / v )%).
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中の各化合物の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表４０に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４０に示す。 About Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment, Nagasal Ogase extract after hydrolyzing treatment, and Nagasal Ogase extract after hydrolyzing and low-temperature storage treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The content of each compound in the Nagasaru Ogase extract after the hydrotreatment was measured after filtering a part of the extract. The results are shown in Table 40. In addition, the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment are also shown in Table 40.
Figure JPOXMLDOC01-appb-T000042
Figure JPOXMLDOC01-appb-T000042
実施例２－５５
　実施例２－５４と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水２．５ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物１２ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１０．５ｇ（蒸発残分０．３００（ｗ／ｖ）％）を得た。 Example 2-55
Digestion was performed by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment and 2.5 mL of distilled water prepared in the same manner as in Example 2-54, and Nagasaga Ogase extraction after hydrolysis was performed. I got a thing.
Further, 12 mL of the Nagasaga Ogase extract after the hydrolysis treatment was stored at −5 ° C. for 5 days and filtered, and then 10.5 g of the Nagasaga Ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.300 (w / v)%) )
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出４１に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４１に示す。 About Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment, Nagasal Ogase extract after hydrolyzing treatment, and Nagasal Ogase extract after hydrolyzing and low-temperature storage treatment, difractic acid content and usnic acid content by the same method as described above Was measured. It shows to Nagasaroogase extraction 41 after a hydration process. In addition, the theoretical value of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment is also shown in Table 41.
Figure JPOXMLDOC01-appb-T000043
Figure JPOXMLDOC01-appb-T000043
実施例２－５６
　実施例２－５４と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水５ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物１４ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１２．６ｇ（蒸発残分０．２４０（ｗ／ｖ）％）を得た。 Example 2-56
Distilled by mixing 10 mL of Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 2-54 and 5 mL of distilled water, and then subjected to hydrolysis. Obtained.
Furthermore, 14 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 5 days and filtered, and 12.6 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (evaporation residue 0.240 (w / v)%) )
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中の各化合物の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表４２に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４２に示す。 About Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment, Nagasal Ogase extract after hydrolyzing treatment, and Nagasal Ogase extract after hydrolyzing and low-temperature storage treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The content of each compound in the Nagasaru Ogase extract after the hydrotreatment was measured after filtering a part of the extract. The results are shown in Table 42. Table 42 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment.
Figure JPOXMLDOC01-appb-T000044
Figure JPOXMLDOC01-appb-T000044
実施例２－５７
　実施例２－５４と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水７．５ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物１６ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１４．１ｇ（蒸発残分０．２００（ｗ／ｖ）％）を得た。 Example 2-57
Digestion was performed by mixing 10 mL of Nagasaga Ogase extract before hydrolysis / low-temperature storage treatment prepared in the same manner as in Example 2-54 and 7.5 mL of distilled water, and extraction was performed after Nagasaga Ogase after hydrolysis. I got a thing.
Further, 16 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 5 days and filtered, and 14.1 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.200 (w / v)%) )
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中の各化合物の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表４３に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４３に示す。 About Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment, Nagasal Ogase extract after hydrolyzing treatment, and Nagasal Ogase extract after hydrolyzing and low-temperature storage treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The content of each compound in the Nagasaru Ogase extract after the hydrotreatment was measured after filtering a part of the extract. The results are shown in Table 43. In addition, the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment are also shown in Table 43.
Figure JPOXMLDOC01-appb-T000045
Figure JPOXMLDOC01-appb-T000045
実施例２－５８
　実施例２－５４と同様にして調製した加水・低温保存処理前のナガサルオガセ抽出物１０ｍＬと蒸留水１０ｍＬとを混合することにより希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物のうち１８ｍＬを－５℃で５日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１５．９ｇ（蒸発残分０．１７０（ｗ／ｖ）％）を得た。 Example 2-58
Distilled by mixing 10 mL of Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment prepared in the same manner as in Example 2-54 and 10 mL of distilled water, and then subjected to hydrolysis. Obtained.
Further, 18 mL of the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 5 days and filtered, and then 15.9 g of the Nagasal Ogase extract after the hydrous and low temperature storage treatment (residual evaporation 0.170 (w / v )%).
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。加水処理後のナガサルオガセ抽出物中の各化合物の含有量の測定は、抽出物の一部をろ過後に測定した。その結果を表４４に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４４に示す。 About Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment, Nagasal Ogase extract after hydrolyzing treatment, and Nagasal Ogase extract after hydrolyzing and low-temperature storage treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The content of each compound in the Nagasaru Ogase extract after the hydrotreatment was measured after filtering a part of the extract. The results are shown in Table 44. In addition, the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the water treatment are also shown in Table 44.
Figure JPOXMLDOC01-appb-T000046
Figure JPOXMLDOC01-appb-T000046
　表４０～４４の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理及び低温保存処理をこの順で行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量を選択的に低減することができる。 As is clear from the results in Tables 40 to 44, with regard to Nagasagaogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent, the hydrolysis treatment and the low-temperature storage treatment were performed in this order. By carrying out in this step, it is possible to selectively reduce the usnic acid content contained in the Nagasaga Ogase extract.
比較例２－１２
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１７０．２４ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。 Comparative Example 2-12
Yokowa Sarugase (obtained from Hichem Corp.) and 10 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 170.24 g (Evaporation residue 0.348 (w / v)) %).
実施例２－５９
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１５６．７９ｇ（蒸発残分０．４０２（ｗ／ｖ）％）を得た。 Example 2-59
Yokowa Sarugase (obtained from Hichem) 10 g and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 156.79 g (evaporation residue 0.402 (w / v)) %).
実施例２－６０
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１６２．１８ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。 Example 2-60
Yokowa Sarugase (obtained from Hichem) 10 g and 50 vol% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 162.18 g (Evaporation residue 0.348 (w / v)) %).
実施例２－６１
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１６６．６１ｇ（蒸発残分０．２５０（ｗ／ｖ）％）を得た。 Example 2-61
Yokowa Sarugase (obtained from Hichem Corp.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days followed by filtration. Yokowa Sarugase extract 166.61 g (evaporation residue 0.250) (W / v)%).
実施例２－６２
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと８０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１７６．５５ｇ（蒸発残分０．２３０（ｗ／ｖ）％）を得た。 Example 2-62
Yokowa Sarugase (obtained from Hichem) and 80 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 176.55 g (evaporation residue 0.230) (W / v)%).
比較例２－１３
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して２０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１４６．４５ｇ（蒸発残分０．４０２（ｗ／ｖ）％）を得た。 Comparative Example 2-13
Yokowa Sarugase (obtained from Hichem) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed and extracted for 4 days at 20 ° C., followed by filtration. Yokowa Sarugase extract 144.65 g (evaporation residue 0.402 (w / v)) %).
比較例２－１４
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１４３．９５ｇ（蒸発残分０．２８８（ｗ／ｖ）％）を得た。 Comparative Example 2-14
Yokowa Sarugase (obtained from Hichem) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 143.95 g (Evaporation residue 0.288 (w / v)) %).
ジフラクタ酸含有量及びウスニン酸含有量の測定
　上記実施例２－５９～２－６２及び比較例２－１２～２－１４で得られた各ヨコワサルオガセ抽出物中のジフラクタ酸含有量及びウスニン酸含有量を、前記と同様の方法により測定した。その結果を表４５に示す。 Measurement of difractic acid content and usnic acid content Difractic acid content and usnic acid content in each Yokowa sucrose extract obtained in Examples 2-59 to 2-62 and Comparative Examples 2-12 to 2-14 The amount was measured by the same method as described above. The results are shown in Table 45.
Figure JPOXMLDOC01-appb-T000047
Figure JPOXMLDOC01-appb-T000047
　表４５の結果から明らかなように、本発明の第２の実施態様の方法によれば、ジフラクタ酸含有量が十分に高くウスニン酸含有量が低減した、ジフラクタ酸高含有かつウスニン酸低含有のヨコワサルオガセ抽出物を得ることができる。これに対して、アルコール含有率の低すぎる抽出溶媒を用いてヨコワサルオガセを抽出すると、ウスニン酸含有量は低下したが、ジフラクタ酸含有量も大きく低下した（比較例２－１２）。また、アルコール含有率の高すぎる抽出溶媒を用いてヨコワサルオガセを抽出すると、ジフラクタ酸含有量は上昇するが、ウスニン酸含有量も上昇した（比較例２－１３及び２－１４）。
　以上のように、サルオガセ属地衣類をアルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出することで、ジフラクタ酸高含有かつウスニン酸低含有のサルオガセ属地衣類抽出物を製造することができる。ウスニン酸は肝毒性を有する一方、ジフラクタ酸は優れたエンドセリン抑制作用を有する。したがって、本発明の第２の実施態様によれば、エンドセリン作用抑制効果を有し安全性に優れたエンドセリン作用抑制剤を製造することができる。 As is apparent from the results in Table 45, according to the method of the second embodiment of the present invention, the difractic acid content was sufficiently high and the usnic acid content was reduced, and the difractic acid content was high and the usnic acid content was low. Yokowa Sarugase extract can be obtained. On the other hand, when Yokosawa ogase was extracted using an extraction solvent having an alcohol content that was too low, the usnic acid content decreased, but the difractic acid content also significantly decreased (Comparative Example 2-12). In addition, when Yokowa sucrose was extracted using an extraction solvent having an alcohol content that was too high, the difractic acid content increased, but the usnic acid content also increased (Comparative Examples 2-13 and 2-14).
As described above, an extract of Salgogase lichen having a high content of difractate and a low content of usnic acid is produced by extracting the Licorice lichen using an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume. be able to. Usnic acid has hepatotoxicity, while difractic acid has an excellent endothelin inhibitory action. Therefore, according to the 2nd embodiment of this invention, the endothelin action inhibitor which has an endothelin action inhibitory effect and was excellent in safety | security can be manufactured.
実施例２－６３
　実施例２－５９で得られた抽出物（加水・低温保存処理前のヨコワサルオガセ抽出物）８８．５７ｇを－５℃で３日間保存後、ろ過することにより低温保存処理後のヨコワサルオガセ抽出物８１．６８ｇ（蒸発残分０．３９０（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のヨコワサルオガセ抽出物８０ｍＬと蒸留水２０ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で４日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物８６．１２ｇ（蒸発残分０．３２８（ｗ／ｖ）％）を得た。 Example 2-63
88.57 g of the extract obtained in Example 2-59 (Yokosawa ogousse extract before hydrolyzing and low-temperature storage treatment) was stored at −5 ° C. for 3 days and filtered to obtain a Yokosawa ogousse extract after low-temperature storage treatment. 81.68 g (evaporation residue 0.390 (w / v)%) was obtained.
80 mL of the obtained Yokosawa-ogase extract after low-temperature storage treatment and 20 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Yokosawa-ogase extract after the hydrotreatment.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at -5 ° C. for 4 days and then filtered, and 86.12 g (Yield residue 0.328 (w / v)%) of the Yokosawa ogase extract after the hydration / low temperature preservation treatment Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、低温保存処理後のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表４６に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４６に示す。 The same method as described above for the Yokosawa Ogase extract before hydrolyzed and cryopreserved, the Yokosawa Ogase extract after the cryopreserved treatment, the Yokosawa Ogase extract after the hydrotreated, and the Yokosawa ogase extract after the hydrolyzed and cryopreserved treatment Was used to measure the difractic acid content and the usnic acid content. The results are shown in Table 46. In addition, the theoretical values of the difractic acid content and the usnic acid content contained in the Yokosawa ogase extract after the water treatment are also shown in Table 46.
Figure JPOXMLDOC01-appb-T000048
Figure JPOXMLDOC01-appb-T000048
実施例２－６４
　実施例２－５９で得られたヨコワサルオガセ抽出物（加水・低温保存処理前のヨコワサルオガセ抽出物）４０ｍＬと蒸留水１０ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で３日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物４１．０３ｇ（蒸発残分０．２８６（ｗ／ｖ）％）を得た。 Example 2-64
40 mL of the Yokosawa Ogase extract obtained in Example 2-59 (Yokosawa Ogase extract before hydrolyzing and low-temperature storage treatment) and 10 mL of distilled water were mixed to dilute the ethanol volume to 40%, followed by hydrolysis. The later Yokosawa Ogase extract was obtained.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at −5 ° C. for 3 days and then filtered, and 41.03 g (Yield residue 0.286 (w / v)%) of the Yokosawa ogase extract after the hydrolyzing and low-temperature preservation treatment Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表４７に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４７に示す。 About Yokosawa-o-gase extract before hydrolyzing and low-temperature preservation treatment, Yokosawa-o-gase extract after hydrolyzing treatment, and Yokosawa-o-gase extract after hydrolyzing and low-temperature preservation treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The results are shown in Table 47. Table 47 also shows the theoretical values of the difractacic acid content and the usnic acid content contained in the Yokowa salugase extract after the water treatment.
Figure JPOXMLDOC01-appb-T000049
Figure JPOXMLDOC01-appb-T000049
実施例２－６５
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、加水・低温保存処理前のヨコワサルオガセ抽出物１６２．１８ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。
　得られた加水・低温保存処理前のヨコワサルオガセ抽出物８０ｍＬと蒸留水２０ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で２日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物８６．９４ｇ（蒸発残分０．３２８（ｗ／ｖ）％）を得た。 Example 2-65
Yokowa Sarugase (obtained from Hichem Corp.) and 50 volume% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. .348 (w / v)%).
80 mL of the obtained Yokosawa-ogase extract before hydrolyzing and low-temperature storage treatment and 20 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain the Yokosawa-ogase extract after the hydrotreatment.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at -5 ° C. for 2 days and then filtered, and 86.94 g (Yield residue 0.328 (w / v)%) of the Yokosawa ogase extract after the hydration / low temperature preservation treatment Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表４８に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表４８に示す。 About Yokosawa-o-gase extract before hydrolyzing and low-temperature preservation treatment, Yokosawa-o-gase extract after hydrolyzing treatment, and Yokosawa-o-gase extract after hydrolyzing and low-temperature preservation treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The results are shown in Table 48. Table 48 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Yokosawa ogousse extract after the water treatment.
Figure JPOXMLDOC01-appb-T000050
Figure JPOXMLDOC01-appb-T000050
　表４６～４８の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるヨコワサルオガセ抽出物について、加水処理及び低温保存処理を行なうことにより、ヨコワサルオガセ抽出物に含まれるジフラクタ酸の含有量を保持したままウスニン酸含有量を選択的に低減することができる。 As is apparent from the results in Tables 46 to 48, the Yokosawa ogase extract obtained by using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a hydrolysis treatment and a low-temperature storage treatment. Thus, the usnic acid content can be selectively reduced while maintaining the content of difractacic acid contained in the Yokosawa ogase extract.
実施例２－６６
　ナガサルオガセ（新和物産社より入手）１０ｇと５体積％エタノール水溶液１Ｌとを混合して２０℃で２９時間処理した後ろ過し、ろ液９６１．９８ｇ（蒸発残分０．０８０（ｗ／ｖ）％）を分離した。次に、ろ液から溶媒を分離して、ナガサルオガセの残渣を得た。
　前記ナガサルオガセの残渣３２．４８ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ナガサルオガセ抽出物１７８．４４ｇ（蒸発残分０．１７０（ｗ／ｖ）％）を得た。 Example 2-66
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of 5 volume% ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 96.98 g of filtrate (evaporation residue 0.080 (w / v )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue.
32.48 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration, and Nagasaga Ogase extract 178.44 g (evaporation residue 0.170 (w / v)%) )
実施例２－６７
　ナガサルオガセ（新和物産社より入手）１０ｇと１体積％エタノール水溶液１Ｌとを混合して２０℃で２９時間処理した後ろ過し、ろ液９５９．０１ｇ（蒸発残分０．０７６（ｗ／ｖ）％）を分離した。次に、ろ液から溶媒を分離して、ナガサルオガセの残渣を得た。
　前記ナガサルオガセの残渣３５．４７ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ナガサルオガセ抽出物１８０．０３ｇ（蒸発残分０．１５２（ｗ／ｖ）％）を得た。 Example 2-67
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of a 1% by volume ethanol aqueous solution were mixed and treated at 20 ° C. for 29 hours, followed by filtration. )%). Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue.
35.47 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days followed by filtration. The Nagasaga Ogase extract 180.03 g (evaporation residue 0.152 (w / v)%) )
実施例２－６８
　ナガサルオガセ（新和物産社より入手）１０ｇと蒸留水１Ｌとを混合して２０℃で２９時間処理した後ろ過し、ろ液９５０．０１ｇ（蒸発残分０．０８０（ｗ／ｖ）％）を分離した。次に、ろ液から溶媒を分離して、ナガサルオガセの残渣を得た。
　前記ナガサルオガセの残渣３２．７５ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ナガサルオガセ抽出物１７９．０３ｇ（蒸発残分０．１４４（ｗ／ｖ）％）を得た。 Example 2-68
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 1 L of distilled water were mixed and treated at 20 ° C. for 29 hours, followed by filtration, and 950.01 g of filtrate (evaporation residue 0.080 (w / v)%) Separated. Next, the solvent was separated from the filtrate to obtain a Nagasaru Ogase residue.
32.75 g of the Nagasaga Ogase residue and 200 mL of a 60% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 5 days, followed by filtration. The Nagasaga Ogase extract 179.03 g (evaporation residue 0.144 (w / v)% )
　上記実施例２－６６～２－６８で得られた各ナガサルオガセ抽出物について、前記と同様の方法によりＨＰＬＣでジフラクタ酸及びウスニン酸含有量を測定した。その結果を表４９に示す。 For each Nagasaruogase extract obtained in Examples 2-66 to 2-68, the contents of difractic acid and usnic acid were measured by HPLC in the same manner as described above. The results are shown in Table 49.
Figure JPOXMLDOC01-appb-T000051
Figure JPOXMLDOC01-appb-T000051
　表４９の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出を行う前に、水系溶媒を用いてサルオガセ属地衣類を処理することにより、ウスニン酸含有量に対してジフラクタ酸含有量が特に高いサルオガセ属地衣類抽出物を得ることができる。 As is clear from the results in Table 49, by performing treatment with Sarhogase lichen using an aqueous solvent before extraction with an aqueous alcohol solution having an alcohol content of 30% by volume to 80% by volume, usnic acid It is possible to obtain an extract of Lichenaceae lichen having a particularly high difractic acid content relative to the content.
実施例２－６９
　実施例２－６６で得られた抽出物（加水・低温保存処理前のナガサルオガセ抽出物）８０ｍＬと蒸留水２０ｍＬとを混合してエタノールの体積が約４０～５０％となるよう希釈して澱出しを行い、加水処理後のナガサルオガセ抽出物を得た。さらに、加水処理後のナガサルオガセ抽出物を－５℃で４日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物８４．３３ｇ（蒸発残分０．１４０（ｗ／ｖ）％）を得た。 Example 2-69
80 mL of the extract obtained in Example 2-66 (Nagasaruogase extract before hydrolyzing and low-temperature storage treatment) and 20 mL of distilled water were mixed to dilute the ethanol volume to about 40 to 50%. The Nagasagaogase extract after the hydration treatment was obtained. Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 4 days and filtered, and 84.33 g of the Nagasao Ogase extract after the hydrous and low temperature preservation treatment (evaporation residue 0.140 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５０に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表５０に示す。 About Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment, Nagasal Ogase extract after hydrolyzing treatment, and Nagasal Ogase extract after hydrolyzing and low-temperature storage treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The results are shown in Table 50. In addition, the theoretical value of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the water treatment is also shown in Table 50.
Figure JPOXMLDOC01-appb-T000052
Figure JPOXMLDOC01-appb-T000052
　表５０の結果から明らかなように、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を抽出溶媒として用いて得られるナガサルオガセ抽出物について、加水処理及び低温保存処理を行なうことにより、ナガサルオガセ抽出物に含まれるウスニン酸含有量を選択的に低減することができる。 As is apparent from the results in Table 50, the Nagasaruogase extract obtained using an aqueous alcohol solution having an alcohol content of 30% by volume or more and 80% by volume or less as an extraction solvent is subjected to a hydration treatment and a low-temperature storage treatment. The usnic acid content contained in the Nagasaga Ogase extract can be selectively reduced.
（第３の実施態様に関する実施例及び比較例）
　以下、第３の実施態様に関する実施例及び比較例について説明する。 (Examples and comparative examples relating to the third embodiment)
Hereinafter, examples and comparative examples relating to the third embodiment will be described.
比較例３－１
　ナガサルオガセ（新和物産社より入手）１０ｇと２０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物２０１．４９ｇ（蒸発残分０．１７２（ｗ／ｖ）％）を得た。 Comparative Example 3-1
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 20 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days, followed by filtration. Nagasaru Ogase extract 201.49 g (evaporation residue 0.172 (w / v)%).
比較例３－２
　ナガサルオガセ（新和物産社より入手）１０ｇと３０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９９．９０ｇ（蒸発残分０．１４６（ｗ／ｖ）％）を得た。 Comparative Example 3-2
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 30 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 199.90 g (evaporation residue 0.146 (w / v)%).
比較例３－３
　ナガサルオガセ（新和物産社より入手）１０ｇと６０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１９８．６９ｇ（蒸発残分０．２４２（ｗ／ｖ）％）を得た。 Comparative Example 3-3
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 60 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaru Ogase extract 198.69 g (evaporation residue 0.242 (w / v)%).
比較例３－４
　ナガサルオガセ（新和物産社より入手）１０ｇと８０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８５．６ｇ（蒸発残分０．２１６（ｗ／ｖ）％）を得た。 Comparative Example 3-4
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 80 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.6 g (evaporation residue 0.216 (w / v)%).
比較例３－５
　ナガサルオガセ（新和物産社より入手）１０ｇと９０体積％エタノール水溶液２００ｍＬとを混合して２０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８３．４ｇ（蒸発残分０．１９２（ｗ／ｖ）％）を得た。 Comparative Example 3-5
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 90 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 7 days followed by filtration. Nagasaga Ogase extract 183.4 g (evaporation residue 0.192 (w / v)%).
比較例３－６
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、ナガサルオガセ抽出物１８５．０６ｇ（蒸発残分０．２６６（ｗ／ｖ）％）を得た。 Comparative Example 3-6
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 40 ° C. for 7 days, filtered, and Nagasaru Ogase extract 185.06 g (evaporation residue 0.266 (w / v)%).
　上記比較例３－１～３－６で得られた各ナガサルオガセ抽出物中のジフラクタ酸含有量及びウスニン酸含有量を、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。結果を表５１に示す。 The difractic acid content and the usnic acid content in each Nagasaruogase extract obtained in Comparative Examples 3-1 to 3-6 were measured for the difractic acid content and the usnic acid content by the same method as described above. . The results are shown in Table 51.
Figure JPOXMLDOC01-appb-T000053
Figure JPOXMLDOC01-appb-T000053
　表５１の結果から明らかなように、抽出溶媒としてアルコール含有率の高いアルコール水溶液を使用すれば、ジフラクタ酸含有量が十分に高いナガサルオガセの抽出物を得ることができる。しかし、その際、ウスニン酸含有量も上昇する（比較例３－３～３－６）。また、アルコール含有率の低い抽出溶媒を用いてナガサルオガセを抽出すると、ウスニン酸含有量は低いがジフラクタ酸含有量も大きく低下してしまう（比較例３－１～３－２）。そのため、得られる抽出物は皮膚外用剤に好適に用いることはできない。 As is apparent from the results in Table 51, when an alcohol aqueous solution having a high alcohol content is used as the extraction solvent, an extract of Nagasaru Ogase with a sufficiently high difractic acid content can be obtained. However, at that time, the usnic acid content also increases (Comparative Examples 3-3 to 3-6). In addition, when Nagasaruogase is extracted using an extraction solvent having a low alcohol content, the usnic acid content is low, but the difractic acid content is greatly reduced (Comparative Examples 3-1 and 3-2). Therefore, the obtained extract cannot be used suitably for a skin external preparation.
比較例３－７
　ナガサルオガセ（新和物産社より入手）５０ｇと９５体積％エタノール水溶液１Ｌとを混合して２０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物７６２．２ｇ（蒸発残分０．３０６（ｗ／ｖ）％）を得た。 Comparative Example 3-7
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 95 L% ethanol aqueous solution 1 L were mixed and extracted at 20 ° C. for 7 days, filtered, and 762.2 g Nagasaga Ogase extract before evaporation and low-temperature storage treatment (evaporation residue) Min 0.306 (w / v)%).
実施例３－１
　比較例３－７で得られた加水・低温保存処理前のナガサルオガセ抽出物８ｍＬと蒸留水１１ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物を－５℃で１２日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１３．５ｇ（蒸発残分０．０８４（ｗ／ｖ）％）を得た。 Example 3-1
The Nagasal Ogase extract obtained by Comparative Example 3-7 before the hydrothermal treatment at low temperature and 8 mL of distilled water and 11 mL of distilled water were mixed to dilute the ethanol volume to 40%. Got.
Furthermore, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 12 days and then filtered, and the Nagasal Ogase extract after the hydrous and low temperature preservation treatment 13.5 g (evaporation residue 0.084 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５２に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表５２に示す。さらに、加水・低温保存処理前のナガサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表５２に示す。 Nagasagaogase extract before hydrolyzing and low-temperature preservation treatment, Nagasaroogase extract after hydrotreating and Nagasaroogase extract after hydrolyzing and low-temperature preservation treatment, difractic acid content and usnic acid content by the same method as described above It was measured. The results are shown in Table 52. Table 52 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment. Furthermore, Table 52 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaru Ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000054
Figure JPOXMLDOC01-appb-T000054
比較例３－８
　ナガサルオガセ（新和物産社より入手）５０ｇと９５体積％エタノール水溶液１Ｌとを混合して４０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物７５９．３ｇ（蒸発残分０．４２０（ｗ／ｖ）％）を得た。 Comparative Example 3-8
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 50 g and 95 L% ethanol aqueous solution 1 L were mixed, extracted for 7 days at 40 ° C., filtered, and then Naga9.3 Ogase extract 759.3 g (evaporation residue) before hydrolyzing and low-temperature storage treatment Min 0.420 (w / v)%).
実施例３－２
　比較例３－８で得られた加水・低温保存処理前のナガサルオガセ抽出物８ｍＬと蒸留水１１ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物を－５℃で１２日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１３．４ｇ（蒸発残分０．１１４（ｗ／ｖ）％）を得た。 Example 3-2
The Nagasal Ogase extract obtained in Comparative Example 3-8 before the hydrolysis / low-temperature storage treatment was mixed with 8 mL of distilled water and 11 mL of distilled water to dilute the ethanol volume to 40%. Got.
Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 12 days and then filtered, and the Nagasal Ogase extract after the hydrous and low temperature preservation treatment 13.4 g (evaporation residue 0.114 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５３に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表５３に示す。さらに、加水・低温保存処理前のナガサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表５３に示す。 Nagasagaogase extract before hydrolyzing and low-temperature preservation treatment, Nagasaroogase extract after hydrotreating and Nagasaroogase extract after hydrolyzing and low-temperature preservation treatment, difractic acid content and usnic acid content by the same method as described above It was measured. The results are shown in Table 53. Table 53 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment. Furthermore, Table 53 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000055
Figure JPOXMLDOC01-appb-T000055
比較例３－９
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍLとを混合して２０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物１４９．１ｇ（蒸発残分０．１６２（ｗ／ｖ）％）を得た。 Comparative Example 3-9
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10g and 95 volume% ethanol aqueous solution 200mL were mixed, extracted at 20 ° C for 7 days, filtered, 149.1g of Nagasaru Ogase extract before hydrolyzing and low-temperature storage treatment (evaporation residue) Min. 0.162 (w / v)%).
実施例３－３
　比較例３－９で得られた加水・低温保存処理前のナガサルオガセ抽出物を－５℃で３７日間保存後、ろ過することにより低温保存処理後のナガサルオガセ抽出物１４０．１ｇ（蒸発残分０．１３６（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のナガサルオガセ抽出物１００ｍＬと蒸留水１３７．５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物を－５℃で６日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物２０６．０ｇ（蒸発残分０．０６（ｗ／ｖ）％）を得た。 Example 3-3
The Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-9 was stored at −5 ° C. for 37 days and then filtered to obtain 140.1 g of Nagasaro Ogase extract after low-temperature storage treatment (evaporation residue). 0.136 (w / v)%) was obtained.
The obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 6 days and then filtered, and the Nagasaga Ogase extract after the hydrous / low temperature preservation treatment 206.0 g (evaporation residue 0.06 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、低温保存処理後のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５４に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の希釈後の理論値も併せて表５４に示す。さらに、加水・低温保存処理前のナガサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表５４に示す。 Nagasaga Ogase extract before hydrolyzed and cryopreserved, Nagasal ogase extract after cryopreserved, Nagasal ogused extract after hydrolyzed, and Nagasal ogused extract after hydrolyzed and cryopreserved Was used to measure the difractic acid content and the usnic acid content. The results are shown in Table 54. Table 54 also shows the theoretical values after dilution of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the hydrotreatment. Furthermore, Table 54 also shows the residual rate of usnic acid relative to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
比較例３－１０
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して３０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物１５０．８ｇ（蒸発残分０．２１８（ｗ／ｖ）％）を得た。 Comparative Example 3-10
Nagasaga Ogase (obtained from Shinwa Bussan Co., Ltd.) 10g and 95 volume% ethanol aqueous solution 200mL were mixed and extracted at 30 ° C for 7 days and filtered. Min 0.218 (w / v)%).
実施例３－４
　比較例３－１０で得られた加水・低温保存処理前のナガサルオガセ抽出物を－５℃で３７日間保存後、ろ過することにより、低温保存処理後のナガサルオガセ抽出物１３７．２ｇ（蒸発残分０．１６６（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のナガサルオガセ抽出物１００ｍＬと蒸留水１３７．５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物を－５℃で６日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物２０４．３ｇ（蒸発残分０．０５（ｗ／ｖ）％）を得た。 Example 3-4
The Nagasaga Ogase extract obtained in Comparative Example 3-10, which had not been hydrolyzed and stored at a low temperature, was stored at −5 ° C. for 37 days and then filtered to obtain 137.2 g of the Nagasal Ogase extract after the low temperature storage (the evaporation residue). Min 0.166 (w / v)%).
The obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 6 days and then filtered, and then 204.3 g of the Nagasaga Ogase extract after the hydrous and low temperature preservation treatment (evaporation residue 0.05 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、低温保存処理後のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５５に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表５５に示す。さらに、加水・低温保存処理前のナガサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表５５に示す。 Nagasaga Ogase extract before hydrolyzed and cryopreserved, Nagasal ogase extract after cryopreserved, Nagasal ogused extract after hydrolyzed, and Nagasal ogused extract after hydrolyzed and cryopreserved Was used to measure the difractic acid content and the usnic acid content. The results are shown in Table 55. In addition, the theoretical value of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment is also shown in Table 55. Furthermore, Table 55 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000057
Figure JPOXMLDOC01-appb-T000057
比較例３－１１
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物１４９．４ｇ（蒸発残分０．２６６（ｗ／ｖ）％）を得た。 Comparative Example 3-11
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed and extracted for 7 days at 40 ° C., followed by filtration. Min 0.266 (w / v)%).
実施例３－５
　比較例３－１１で得られた加水・低温保存処理前のナガサルオガセ抽出物を－５℃で３３日間保存後、ろ過することにより、低温保存処理後のナガサルオガセ抽出物１３９．７ｇ（蒸発残分０．１９６（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のナガサルオガセ抽出物１００ｍＬと蒸留水１３７．５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物を－５℃で３日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物２０５．７ｇ（蒸発残分０．０６（ｗ／ｖ）％）を得た。 Example 3-5
The Nagasaga Ogase extract obtained in Comparative Example 3-11 before the hydrothermal and low temperature preservation treatment was stored at −5 ° C. for 33 days and filtered to obtain 139.7 g of the Nagasal Ogase extract after the low temperature preservation treatment (evaporation residue). Min 0.196 (w / v)%).
The obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 3 days and then filtered, and the Nagasaga Ogase extract after the hydrous / low temperature preservation treatment 205.7 g (evaporation residue 0.06 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、低温保存処理後のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５６に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表５６に示す。さらに、加水・低温保存処理前のナガサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表５６に示す。 Nagasaroogase extract before hydrolyzing and low-temperature preservation treatment, Nagasaroogase extract after cryopreservation treatment, Nagasaroogase extract after hydrolyzing treatment, and Nagasaroogase extract after hydrolyzing and cryopreservation treatment Was used to measure the difractic acid content and the usnic acid content. The results are shown in Table 56. In addition, Table 56 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment. Furthermore, Table 56 also shows the residual rate of usnic acid relative to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000058
Figure JPOXMLDOC01-appb-T000058
比較例３－１２
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して２０℃で３日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物１４６．１４ｇ（蒸発残分０．２６６（ｗ／ｖ）％）を得た。 Comparative Example 3-12
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed, extracted at 20 ° C. for 3 days, filtered, and then 146.14 g (Evaporation residue) Min 0.266 (w / v)%).
実施例３－６
　比較例３－１２で得られた加水・低温保存処理前のナガサルオガセ抽出物１３９．６２ｇを－５℃で３日間保存後、ろ過することにより、低温保存処理後のナガサルオガセ抽出物１３３．６１ｇ（蒸発残分０．２４８（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のナガサルオガセ抽出物１００ｍＬと蒸留水１３７．５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物を－５℃で３日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物２０１．５８ｇ（蒸発残分０．０９（ｗ／ｖ）％）を得た。 Example 3-6
133.61 g of Nagasaga Ogase extract after low-temperature storage treatment was filtered by storing 139.62 g of Nagasaro-ogase extract before hydrolysis / low-temperature storage treatment obtained in Comparative Example 3-12 at -5 ° C. for 3 days. (Evaporation residue 0.248 (w / v)%) was obtained.
The obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 3 days and then filtered, and 201.58 g of the Nagasal Ogase extract after the hydrothermal treatment at low temperature (evaporation residue 0.09 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、低温保存処理後のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５７に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表５７に示す。さらに、加水・低温保存処理前のナガサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表５７に示す。 Nagasaga Ogase extract before hydrolyzed and cryopreserved, Nagasal ogase extract after cryopreserved, Nagasal ogused extract after hydrolyzed, and Nagasal ogused extract after hydrolyzed and cryopreserved Was used to measure the difractic acid content and the usnic acid content. The results are shown in Table 57. Table 57 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaruogase extract after the hydrolysis treatment. Furthermore, Table 57 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000059
Figure JPOXMLDOC01-appb-T000059
比較例３－１３
　ナガサルオガセ（新和物産社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で７日間抽出後ろ過し、加水・低温保存処理前のナガサルオガセ抽出物１４３．５２ｇ（蒸発残分０．４１０（ｗ／ｖ）％）を得た。 Comparative Example 3-13
Nagasaru Ogase (obtained from Shinwa Bussan Co., Ltd.) and 95% ethanol aqueous solution (200 mL) were mixed and extracted at 40 ° C. for 7 days followed by filtration. Min. 0.410 (w / v)%).
実施例３－７
　比較例３－１３で得られた加水・低温保存処理前のナガサルオガセ抽出物１４１．５０ｇを－５℃で２日間保存後、ろ過することにより、低温保存処理後のナガサルオガセ抽出物１３２．３１ｇ（蒸発残分０．３５８（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のナガサルオガセ抽出物１００ｍＬと蒸留水１３７．５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のナガサルオガセ抽出物を得た。
　さらに、加水処理後のナガサルオガセ抽出物を－５℃で３日間保存後ろ過し、加水・低温保存処理後のナガサルオガセ抽出物１９７．０ｇ（蒸発残分０．１０４（ｗ／ｖ）％）を得た。 Example 3-7
141.50 g of the Nagasaga Ogase extract before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-13 was stored at −5 ° C. for 2 days and filtered to obtain 132.31 g of Nagasaga Ogase extract after low-temperature storage treatment. (Evaporation residue 0.358 (w / v)%) was obtained.
The obtained Nagasaga Ogase extract after the low-temperature storage treatment and 137.5 mL of distilled water were mixed to dilute the ethanol volume to 40% to obtain a Nagasaga Ogase extract after the hydrotreatment.
Further, the Nagasaga Ogase extract after the hydrotreatment was stored at −5 ° C. for 3 days and filtered, and 197.0 g of the Nagasal Ogase extract after the hydrous and low-temperature storage treatment (evaporation residue 0.104 (w / v)%) Got.
　加水・低温保存処理前のナガサルオガセ抽出物、低温保存処理後のナガサルオガセ抽出物、加水処理後のナガサルオガセ抽出物、及び加水・低温保存処理後のナガサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表５８に示す。なお、加水処理後のナガサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表５８に示す。さらに、加水・低温保存処理前のナガサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表５８に示す。 Nagasaga Ogase extract before hydrolyzed and cryopreserved, Nagasal ogase extract after cryopreserved, Nagasal ogused extract after hydrolyzed, and Nagasal ogused extract after hydrolyzed and cryopreserved Was used to measure the difractic acid content and the usnic acid content. The results are shown in Table 58. In addition, Table 58 also shows the theoretical values of the difractic acid content and the usnic acid content contained in the Nagasaroogase extract after the hydrotreatment. Furthermore, Table 58 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Nagasaga Ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000060
Figure JPOXMLDOC01-appb-T000060
　表５２～５８の結果から明らかなように、アルコール含有率が７０体積％を超えるアルコール水溶液を抽出溶媒として用いてサルオガセ属地衣類を抽出物しただけでは、抽出物中のウスニン酸含有量は非常に高い。これに対して、ウスニン酸の選択的低減処理を行なうことにより、ウスニン酸含有量を選択的に低減することができる。さらに、サルオガセ属地衣類抽出物に含まれるジフラクタ酸の含有量をある程度保持したままウスニン酸含有量を選択的に低減することができ、ウスニン酸の選択的低減処理前の抽出物と比較した場合、ジフラクタ酸濃度がウスニン酸濃度よりもさらに高くなった抽出物が得られる。 As is apparent from the results in Tables 52 to 58, the extract of lichenaceae lichen using only an alcohol aqueous solution with an alcohol content of more than 70% by volume as the extraction solvent has a very high usnic acid content in the extract. high. On the other hand, the usnic acid content can be selectively reduced by performing a selective reduction process of usnic acid. Furthermore, it is possible to selectively reduce the content of usnic acid while maintaining the content of difractic acid contained in the extract of the genus Sarhogase to some extent, when compared with the extract before the selective reduction treatment of usnic acid, An extract with a difractic acid concentration higher than the usnic acid concentration is obtained.
比較例３－１４
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと１０体積％エタノール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１７０．２４ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。 Comparative Example 3-14
Yokowa Sarugase (obtained from Hichem Corp.) and 10 volume% ethanol aqueous solution 200 mL were mixed and extracted at 20 ° C. for 5 days, followed by filtration. Yokowa Sarugase extract 170.24 g (Evaporation residue 0.348 (w / v)) %).
比較例３－１５
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１６２．１８ｇ（蒸発残分０．３４８（ｗ／ｖ）％）を得た。 Comparative Example 3-15
Yokowa Sarugase (obtained from Hichem) 10 g and 50 vol% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 162.18 g (Evaporation residue 0.348 (w / v)) %).
比較例３－１６
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと５０体積％1,3-ブタンジオール水溶液２００ｍＬとを混合して２０℃で５日間抽出後ろ過し、ヨコワサルオガセ抽出物１６６．６１ｇ（蒸発残分０．２５０（ｗ／ｖ）％）を得た。 Comparative Example 3-16
Yokowa Sarugase (obtained from Hichem Corp.) and 50 volume% 1,3-butanediol aqueous solution (200 mL) were mixed and extracted at 20 ° C. for 5 days followed by filtration. Yokowa Sarugase extract 166.61 g (evaporation residue 0.250) (W / v)%).
比較例３－１７
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して２０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１４６．４５ｇ（蒸発残分０．４０２（ｗ／ｖ）％）を得た。 Comparative Example 3-17
Yokowa Sarugase (obtained from Hichem) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed and extracted for 4 days at 20 ° C., followed by filtration. Yokowa Sarugase extract 144.65 g (evaporation residue 0.402 (w / v)) %).
比較例３－１８
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、ヨコワサルオガセ抽出物１４３．９５ｇ（蒸発残分０．２８８（ｗ／ｖ）％）を得た。 Comparative Example 3-18
Yokowa Sarugase (obtained from Hichem) 10 g and 95 volume% ethanol aqueous solution 200 mL were mixed and extracted at 40 ° C. for 4 days followed by filtration. Yokowa Sarugase extract 143.95 g (Evaporation residue 0.288 (w / v)) %).
ジフラクタ酸濃度及びウスニン酸濃度の測定
　上記比較例３－１４～３－１８で得られた各ヨコワサルオガセ抽出物中のジフラクタ酸含有量及びウスニン酸含有量を、前記と同様の方法により測定した。その結果を表５９に示す。 Measurement of Difractic Acid Concentration and Usnic Acid Concentration The difractic acid content and the usnic acid content in each Yokowa sucrose extract obtained in Comparative Examples 3-14 to 3-18 were measured by the same method as described above. The results are shown in Table 59.
Figure JPOXMLDOC01-appb-T000061
Figure JPOXMLDOC01-appb-T000061
　表５９の結果から明らかなように、抽出溶媒としてアルコール含有率の高いアルコール水溶液を使用すれば、ジフラクタ酸含有量が十分に高いヨコワサルオガセの抽出物を得ることができる。しかし、その際、ウスニン酸含有量も上昇する（比較例３－１５、３－１７及び３－１８）。また、アルコール含有率の低い抽出溶媒を用いてナガサルオガセを抽出すると、ウスニン酸含有量は低いがジフラクタ酸含有量も大きく低下してしまう（比較例３－１４及び３－１６）。そのため、得られる抽出物は皮膚外用剤に好適に用いることはできない。 As is apparent from the results in Table 59, when an alcohol aqueous solution having a high alcohol content is used as the extraction solvent, an extract of Yokosawa ogase having a sufficiently high difractic acid content can be obtained. However, at that time, the usnic acid content also increases (Comparative Examples 3-15, 3-17 and 3-18). In addition, when Nagasaruogase is extracted using an extraction solvent having a low alcohol content, the usnic acid content is low, but the difractic acid content is greatly reduced (Comparative Examples 3-14 and 3-16). Therefore, the obtained extract cannot be used suitably for a skin external preparation.
実施例３－８
　比較例３－１７で得られた抽出物（加水・低温保存処理前のヨコワサルオガセ抽出物）８１．０５ｇを－５℃で３日間保存後、ろ過することにより、低温保存処理後のヨコワサルオガセ抽出物７５．０２ｇ（蒸発残分０．２７２（ｗ／ｖ）％）を得た。
　得られた低温保存処理後のヨコワサルオガセ抽出物５０ｍＬと蒸留水６８．７５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で４日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物９８．９７ｇ（蒸発残分０．０８（ｗ／ｖ）％）を得た。 Example 3-8
Extract 81.05 g of the extract obtained in Comparative Example 3-17 (Yokosawa-o-gase extract before hydrolyzing and low-temperature storage treatment) was stored at −5 ° C. for 3 days, and then filtered to extract Yokosawa-o-gase after low-temperature storage treatment. As a result, 75.02 g (0.272 (w / v)% of evaporation residue) of the product was obtained.
The obtained Yokosawa ogase extract after low-temperature storage treatment and 68.75 mL of distilled water were mixed and diluted so that the volume of ethanol was 40%, and the Yokosawa ogase extract after the hydration treatment was obtained.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at −5 ° C. for 4 days and then filtered, and 98.97 g of the Yokosawa ogase extract after the hydrothermal treatment at low temperature (evaporation residue 0.08 (w / v)%) Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、低温保存処理後のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表６０に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表６０に示す。さらに、加水・低温保存処理前のヨコワサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表６０に示す。 The same method as described above for the Yokosawa Ogase extract before hydrolyzed and cryopreserved, the Yokosawa Ogase extract after the cryopreserved treatment, the Yokosawa Ogase extract after the hydrotreated, and the Yokosawa ogase extract after the hydrolyzed and cryopreserved treatment Was used to measure the difractic acid content and the usnic acid content. The results are shown in Table 60. Table 60 also shows the theoretical values of the difractacic acid content and the usnic acid content contained in the Yokosawa ogase extract after the water treatment. Furthermore, Table 60 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Yokosawa-ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000062
Figure JPOXMLDOC01-appb-T000062
比較例３－１９
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して２０℃で４日間抽出後ろ過し、加水・低温保存処理前のヨコワサルオガセ抽出物１４６．４５ｇ（蒸発残分０．４０２（ｗ／ｖ）％）を得た。 Comparative Example 3-19
10 g of Yokowa Salugase (obtained from Hichem) and 200 mL of a 95% by volume ethanol aqueous solution were mixed and extracted at 20 ° C. for 4 days, followed by filtration, and 146.45 g of Yokowa Salugase extract before hydrolyzing and low-temperature storage treatment (0% evaporation residue). .402 (w / v)%).
実施例３－９
　比較例３－１９で得られた加水・低温保存処理前のヨコワサルオガセ抽出物５０ｍＬと蒸留水６８．７５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で２日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物１０１．１５ｇ（蒸発残分０．０６２（ｗ／ｖ）％）を得た。 Example 3-9
50 ml of the extract of Yokosawa-o-gase before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-19 and 68.75 mL of distilled water were mixed to dilute the ethanol volume to 40%. An extract was obtained.
Further, the Yokosawa ogase extract after the hydrolysis treatment was stored at −5 ° C. for 2 days and filtered, and 101.15 g of the Yokosawa ogase extract after the hydrolysis / low temperature preservation treatment (evaporation residue 0.062 (w / v)%) Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表６１に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表６１に示す。さらに、加水・低温保存処理前のヨコワサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表６１に示す。 About Yokosawa-o-gase extract before hydrolyzing and low-temperature preservation treatment, Yokosawa-o-gase extract after hydrolyzing treatment, and Yokosawa-o-gase extract after hydrolyzing and low-temperature preservation treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The results are shown in Table 61. In addition, the theoretical value of the difractic acid content and the usnic acid content contained in the Yokosawa ogase extract after the hydrotreatment is also shown in Table 61. Furthermore, Table 61 also shows the residual rate of usnic acid with respect to the usnic acid content contained in the Yokosawa-o-gase extract before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000063
Figure JPOXMLDOC01-appb-T000063
比較例３－２０
　ヨコワサルオガセ（ハイケム社より入手）１０ｇと９５体積％エタノール水溶液２００ｍＬとを混合して４０℃で４日間抽出後ろ過し、加水・低温保存処理前のヨコワサルオガセ抽出物１４３．９５ｇ（蒸発残分０．２８８（ｗ／ｖ）％）を得た。 Comparative Example 3-20
Yokowa Sarugase (obtained from Hichem Corp.) and 95 volume% aqueous ethanol solution (200 mL) were mixed and extracted at 40 ° C. for 4 days, followed by filtration, and 143.95 g (Yield residue 0) 288 (w / v)%).
実施例３－１０
　比較例３－２０で得られた加水・低温保存処理前のヨコワサルオガセ抽出物５０ｍＬと蒸留水６８．７５ｍＬとを混合してエタノールの体積が４０％となるよう希釈し、加水処理後のヨコワサルオガセ抽出物を得た。
　さらに、加水処理後のヨコワサルオガセ抽出物を－５℃で２日間保存後ろ過し、加水・低温保存処理後のヨコワサルオガセ抽出物１００．１５ｇ（蒸発残分０．１０２（ｗ／ｖ）％）を得た。 Example 3-10
50 mL of the extract of Yokosawa-o-gase before hydrolyzing and low-temperature storage treatment obtained in Comparative Example 3-20 and 68.75 mL of distilled water were mixed to dilute the ethanol volume to 40%. An extract was obtained.
Further, the Yokosawa ogase extract after the hydrotreatment was stored at −5 ° C. for 2 days and then filtered, and 100.15 g of the Yokosawa ogase extract after the hydrothermal treatment at low temperature (evaporation residue 0.102 (w / v)%) Got.
　加水・低温保存処理前のヨコワサルオガセ抽出物、加水処理後のヨコワサルオガセ抽出物、及び加水・低温保存処理後のヨコワサルオガセ抽出物について、前記と同様の方法によりジフラクタ酸含有量及びウスニン酸含有量を測定した。その結果を表６２に示す。なお、加水処理後のヨコワサルオガセ抽出物に含まれるジフラクタ酸含有量及びウスニン酸含有量の理論値も併せて表６２に示す。さらに、加水・低温保存処理前のヨコワサルオガセ抽出物に含まれるウスニン酸含有量に対するウスニン酸の残存率も表６２に示す。 About Yokosawa-o-gase extract before hydrolyzing and low-temperature preservation treatment, Yokosawa-o-gase extract after hydrolyzing treatment, and Yokosawa-o-gase extract after hydrolyzing and low-temperature preservation treatment, difractic acid content and usnic acid content by the same method as described above Was measured. The results are shown in Table 62. In addition, the theoretical value of the difractic acid content and the usnic acid content contained in the Yokosawa ogase extract after the water treatment is also shown in Table 62. Furthermore, Table 62 also shows the residual rate of usnic acid with respect to the content of usnic acid contained in the extract of Yokosawa-ogase before the hydrothermal treatment at low temperature.
Figure JPOXMLDOC01-appb-T000064
Figure JPOXMLDOC01-appb-T000064
　表６０～６２の結果から明らかなように、アルコール含有率が７０体積％を超えるアルコール水溶液を抽出溶媒として用いてサルオガセ属地衣類を抽出物しただけでは、抽出物中のウスニン酸含有量は非常に高い。これに対して、ウスニン酸の選択的低減処理を行なうことにより、ウスニン酸含有量を選択的に低減することができる。さらに、サルオガセ属地衣類抽出物に含まれるジフラクタ酸の含有量をある程度保持したままウスニン酸含有量を選択的に低減することができ、ウスニン酸の選択的低減処理前の抽出物と比較した場合、ジフラクタ酸濃度がウスニン酸濃度よりもさらに高くなった抽出物が得られる。 As is clear from the results in Tables 60 to 62, the extract of lichenaceae lichen using only an alcohol aqueous solution with an alcohol content of more than 70% by volume as the extraction solvent has a very high usnic acid content in the extract. high. On the other hand, the usnic acid content can be selectively reduced by performing a selective reduction process of usnic acid. Furthermore, it is possible to selectively reduce the content of usnic acid while maintaining the content of difractic acid contained in the extract of the genus Sarhogase to some extent, when compared with the extract before the selective reduction treatment of usnic acid, An extract with a difractic acid concentration higher than the usnic acid concentration is obtained.
（処方例）
　前記実施例で得られた抽出物を有効成分とする、下記に示す組成のローション、乳液、美容液及びクリームの形態の美白剤を常法により各々調製出来る。 (Prescription example)
Whitening agents in the form of lotions, emulsions, cosmetic liquids and creams having the composition shown below, each of which contains the extract obtained in the above example as an active ingredient, can be prepared by conventional methods.
１．ローションの調製
（組成）　　　　　　　　　　　　　　　　　　　　（配合：質量％）
1,3-ブチレングリコール　　　　　　　　　　　　　　　８．０
グリセリン　　　　　　　　　　　　　　　　　　　　　５．０
エタノール　　　　　　　　　　　　　　　　　　　　　３．０
実施例１－９、実施例２－５又は実施例３－１
　で得られたナガサルオガセ抽出物　　　　　　　　　　３．０
カミツレエキス　　　　　　　　　　　　　　　　　　　３．０
キキョウエキス　　　　　　　　　　　　　　　　　　　１．０
チョウジエキス　　　　　　　　　　　　　　　　　　　１．０
キサンタンガム　　　　　　　　　　　　　　　　　　　０．１
ヒアルロン酸　　　　　　　　　　　　　　　　　　　　０．１
リン酸二ナトリウム　　　　　　　　　　　　　　　　　０．１
リン酸二水素ナトリウム　　　　　　　　　　　　　　　０．１
アスコルビン酸2-グルコシド　　　　　　　　　　　　　２．０
精製水　　　　　　　　　　　　　　　　　　　　　　　残部
香料　　　　　　　　　　　　　　　　　　　　　　　　適量
防腐剤　　　　　　　　　　　　　　　　　　　　　　　適量 1. Preparation of lotion (composition) (Formulation: mass%)
1,3-butylene glycol 8.0
Glycerin 5.0
Ethanol 3.0
Example 1-9, Example 2-5 or Example 3-1.
Nagasaroogase extract obtained in 3.0
Chamomile extract 3.0
Kyokyo Extract 1.0
Clove extract 1.0
Xanthan gum 0.1
Hyaluronic acid 0.1
Disodium phosphate 0.1
Sodium dihydrogen phosphate 0.1
Ascorbic acid 2-glucoside 2.0
Purified water Remaining fragrance Appropriate amount Preservative Appropriate amount
２．乳液の調製
（組成）　　　　　　　　　　　　　　　　　　　　（配合：質量％）
実施例１－１０、実施例２－６又は実施例３－２
　で得られたナガサルオガセ抽出物　　　　　　　　　　１．０
カミツレエキス　　　　　　　　　　　　　　　　　　　１．０
キキョウエキス　　　　　　　　　　　　　　　　　　　１．０
アルテアエキス　　　　　　　　　　　　　　　　　　　２．０
スクワラン　　　　　　　　　　　　　　　　　　　　　３．０
オリブ油　　　　　　　　　　　　　　　　　　　　　　３．０
グリセリン　　　　　　　　　　　　　　　　　　　　　５．０
ポリオキシエチレン硬化ヒマシ油
　（エチレンオキサイドの付加モル数：40）　　　　　　１．０
カルボキシビニルポリマー　　　　　　　　　　　　　　０．２
水酸化カリウム　　　　　　　　　　　　　　　　　　　０．１
キサンタンガム　　　　　　　　　　　　　　　　　　　０．２
エデト酸二ナトリウム　　　　　　　　　　　　　　　　０．０２
精製水　　　　　　　　　　　　　　　　　　　　　　　残部
防腐剤　　　　　　　　　　　　　　　　　　　　　　　適量 2. Preparation of milky lotion (composition) (formulation: mass%)
Example 1-10, Example 2-6 or Example 3-2
Nagasaruogase extract obtained in 1.0
Chamomile extract 1.0
Kyokyo Extract 1.0
Altea extract 2.0
Squalane 3.0
Olive oil 3.0
Glycerin 5.0
Polyoxyethylene hydrogenated castor oil (number of moles of ethylene oxide added: 40) 1.0
Carboxyvinyl polymer 0.2
Potassium hydroxide 0.1
Xanthan gum 0.2
Edetate disodium 0.02
Purified water Remaining antiseptic
３．美容液の調製
（組成）　　　　　　　　　　　　　　　　　　　　（配合：質量％）
実施例１－１１、実施例２－７又は実施例３－３
　で得られたナガサルオガセ抽出物　　　　　　　　　　１．０
カミツレエキス　　　　　　　　　　　　　　　　　　　１．０
キキョウエキス　　　　　　　　　　　　　　　　　　　１．０
チョウジエキス　　　　　　　　　　　　　　　　　　　１．０
カルボキシビニルポリマー　　　　　　　　　　　　　　０．２
アクリル酸・メタクリル酸アルキル共重合体　　　　　　０．２
水酸化カリウム　　　　　　　　　　　　　　　　　　　０．２
キサンタンガム　　　　　　　　　　　　　　　　　　　０．１
ヒアルロン酸　　　　　　　　　　　　　　　　　　　　０．２
クエン酸ナトリウム　　　　　　　　　　　　　　　　　０．１５
クエン酸　　　　　　　　　　　　　　　　　　　　　　０．０３
グリセリン　　　　　　　　　　　　　　　　　　　　　１０．０
1,3-ブチレングリコール　　　　　　　　　　　　　　　５．０
エデト酸二ナトリウム　　　　　　　　　　　　　　　　０．０５
精製水　　　　　　　　　　　　　　　　　　　　　　　残部
防腐剤　　　　　　　　　　　　　　　　　　　　　　　適量
香料　　　　　　　　　　　　　　　　　　　　　　　　適量 3. Preparation of serum (composition) (Composition: mass%)
Example 1-11, Example 2-7 or Example 3-3
Nagasaruogase extract obtained in 1.0
Chamomile extract 1.0
Kyokyo Extract 1.0
Clove extract 1.0
Carboxyvinyl polymer 0.2
Acrylic acid / alkyl methacrylate copolymer 0.2
Potassium hydroxide 0.2
Xanthan gum 0.1
Hyaluronic acid 0.2
Sodium citrate 0.15
Citric acid 0.03
Glycerin 10.0
1,3-butylene glycol 5.0
Edetate disodium 0.05
Purified water Remaining preservative Appropriate amount Perfume Appropriate amount
４．美容液の調製
（組成）　　　　　　　　　　　　　　　　　　　　（配合：質量％）
実施例１－６、実施例２－２又は実施例３－５
　で得られたナガサルオガセ抽出物　　　　　　　　　　３．０
カミツレエキス　　　　　　　　　　　　　　　　　　　１．０
チョウジエキス　　　　　　　　　　　　　　　　　　　１．０
キキョウエキス　　　　　　　　　　　　　　　　　　　１．０
キサンタンガム　　　　　　　　　　　　　　　　　　　０．２
カルボキシメチルセルロース　　　　　　　　　　　　　０．２
カルボキシビニルポリマー　　　　　　　　　　　　　　０．２
水酸化カリウム　　　　　　　　　　　　　　　　　　　０．１
クエン酸　　　　　　　　　　　　　　　　　　　　　　０．０３
クエン酸ナトリウム　　　　　　　　　　　　　　　　　０．１５
グリセリン　　　　　　　　　　　　　　　　　　　　　５．０
プロピレングリコール　　　　　　　　　　　　　　　　３．０
ポリエチレングリコール（分子量１５００）　　　　　　１．０
モノステアリン酸ポリエチレングリコール　　　　　　　０．５
精製水　　　　　　　　　　　　　　　　　　　　　　　残部
防腐剤　　　　　　　　　　　　　　　　　　　　　　　適量 4). Preparation of serum (composition) (Composition: mass%)
Example 1-6, Example 2-2 or Example 3-5
Nagasaroogase extract obtained in 3.0
Chamomile extract 1.0
Clove extract 1.0
Kyokyo Extract 1.0
Xanthan gum 0.2
Carboxymethylcellulose 0.2
Carboxyvinyl polymer 0.2
Potassium hydroxide 0.1
Citric acid 0.03
Sodium citrate 0.15
Glycerin 5.0
Propylene glycol 3.0
Polyethylene glycol (molecular weight 1500) 1.0
Polyethylene glycol monostearate 0.5
Purified water Remaining antiseptic
５．クリームの調製
（組成）　　　　　　　　　　　　　　　　　　　　（配合：質量％）
実施例１－１２、実施例２－８又は実施例３－１
　で得られたナガサルオガセ抽出物　　　　　　　　　　３．０
カミツレエキス　　　　　　　　　　　　　　　　　　　２．０
キキョウエキス　　　　　　　　　　　　　　　　　　　２．０
チョウジエキス　　　　　　　　　　　　　　　　　　　２．０
メチルポリシロキサン　　　　　　　　　　　　　　　　３．０
スクワラン　　　　　　　　　　　　　　　　　　　　　２．０
ジカプリン酸ネオペンチルグリコール　　　　　　　　　３．０
ステアリルアルコール　　　　　　　　　　　　　　　　１．５
セタノール　　　　　　　　　　　　　　　　　　　　　１．０
ポリオキシエチレン硬化ヒマシ油
　（エチレンオキサイドの付加モル数：60）　　　　　　０．５
アクリル酸・メタクリル酸アルキル共重合体　　　　　　０．３
水酸化カリウム　　　　　　　　　　　　　　　　　　　０．１５
キサンタンガム　　　　　　　　　　　　　　　　　　　０．１
エデト酸二ナトリウム　　　　　　　　　　　　　　　　０．０５
精製水　　　　　　　　　　　　　　　　　　　　　　　残部
防腐剤　　　　　　　　　　　　　　　　　　　　　　　適量
香料　　　　　　　　　　　　　　　　　　　　　　　　適量 5. Preparation (composition) of cream (formulation: mass%)
Example 1-12, Example 2-8 or Example 3-1.
Nagasaroogase extract obtained in 3.0
Chamomile extract 2.0
Kyokyo Extract 2.0
Clove extract 2.0
Methylpolysiloxane 3.0
Squalane 2.0
Neopentyl glycol dicaprate 3.0
Stearyl alcohol 1.5
Cetanol 1.0
Polyoxyethylene hydrogenated castor oil (Mole number of ethylene oxide added: 60) 0.5
Acrylic acid / alkyl methacrylate copolymer 0.3
Potassium hydroxide 0.15
Xanthan gum 0.1
Edetate disodium 0.05
Purified water Remaining preservative Appropriate amount Perfume Appropriate amount
　本発明をその実施態様とともに説明したが、我々は特に指定しない限り我々の発明を説明のどの細部においても限定しようとするものではなく、添付の請求の範囲に示した発明の精神と範囲に反することなく幅広く解釈されるべきであると考える。 While this invention has been described in conjunction with its embodiments, we do not intend to limit our invention in any detail of the description unless otherwise specified and are contrary to the spirit and scope of the invention as set forth in the appended claims. I think it should be interpreted widely.
　本願は、2012年6月8日に日本国で特許出願された特願2012-130797、2012年6月8日に日本国で特許出願された特願2012-130798、及び2012年6月8日に日本国で特許出願された特願2012-130799に基づく優先権を主張するものであり、これらはここに参照してその内容を本明細書の記載の一部として取り込む。 This application includes Japanese Patent Application No. 2012-130797 filed in Japan on June 8, 2012, Japanese Patent Application No. 2012-130798 filed in Japan on June 8, 2012, and June 8, 2012. Claiming priority based on Japanese Patent Application No. 2012-130799 filed in Japan, which is hereby incorporated herein by reference in its entirety.

Claims (26)

  1. 　サルオガセ（Usnea）属地衣類を、アルコール含有率が８０体積％以下のアルコール水溶液を用いて抽出する、ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。 A method for producing an extract containing Salusose genus liquor having a low content of usnic acid, wherein a lichen belonging to the genus Usnea is extracted using an alcohol aqueous solution having an alcohol content of 80% by volume or less.
  2. 　サルオガセ（Usnea）属地衣類を、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出する、ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。 Usnea the (Usnea) territorial clothing manufacturing method of the alcohol content is extracted using the following aqueous alcohol 80 vol% to 30 vol%, Jifurakuta acid-rich and usnic acid low content Usnea territorial garment extract.
  3. 　前記アルコール水溶液がエタノール水溶液又は1,3-ブタンジオール水溶液である、請求項１又は２記載の製造方法。 3. The production method according to claim 1 or 2, wherein the alcohol aqueous solution is an ethanol aqueous solution or a 1,3-butanediol aqueous solution.
  4. 　前記サルオガセ属地衣類がナガサルオガセ（Usnea　longissima）又はヨコワサルオガセ（Usnea　diffracta　Vain.）である、請求項１～３のいずれか１項記載の製造方法。 The production method according to any one of claims 1 to 3, wherein the lichen belonging to the genus Sargogase is Usa longissima or Usnea diffracta Vain.
  5. 　抽出したサルオガセ属地衣類抽出物に対して、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理及び加熱処理からなる群より選ばれる１種以上の処理によりウスニン酸の低減処理を行う、請求項１、３及び４のいずれか１項記載の製造方法。 The extracted sarcoma lichen extract is subjected to a reduction treatment of usnic acid by one or more treatments selected from the group consisting of low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment and heat treatment. The manufacturing method of any one of 3 and 4.
  6. 　抽出したサルオガセ属地衣類抽出物に対して、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理及び加熱処理からなる群より選ばれる１種以上の処理によりウスニン酸の選択的低減処理を行う、請求項２～４のいずれか１項記載の製造方法。 The extracted sarcoma lichen extract is subjected to a selective reduction treatment of usnic acid by one or more treatments selected from the group consisting of low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment, and heat treatment. Item 5. The production method according to any one of Items 2 to 4.
  7. 　水系溶媒を用いて前記サルオガセ属地衣類を処理し、処理したサルオガセ属地衣類に対して前記アルコール水溶液を用いた抽出を行う、請求項１～６のいずれか１項記載の製造方法。 The production method according to any one of claims 1 to 6, wherein the Saruogase lichen is treated with an aqueous solvent, and the treated Saruogase lichen is extracted with the aqueous alcohol solution.
  8. 　得られた抽出物中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が３以上である、請求項２～４及び６のいずれか１項記載の製造方法。 The production method according to any one of claims 2 to 4 and 6, wherein the content of difractic acid is 3 or more when the content of usnic acid in the obtained extract is 1 on a mass basis.
  9. 　サルオガセ（Usnea）属地衣類を、アルコール含有率が３０体積％以上８０体積％以下のアルコール水溶液を用いて抽出する、エンドセリン作用抑制剤の製造方法。 A method for producing an endothelin action inhibitor, wherein a lichen belonging to the genus Usnea is extracted using an alcohol aqueous solution having an alcohol content of 30% by volume to 80% by volume.
  10. 　前記アルコール水溶液がエタノール水溶液又は1,3-ブタンジオール水溶液である、請求項９記載の製造方法。 The production method according to claim 9, wherein the alcohol aqueous solution is an ethanol aqueous solution or a 1,3-butanediol aqueous solution.
  11. 　前記サルオガセ属地衣類がナガサルオガセ（Usnea　longissima）又はヨコワサルオガセ（Usnea　diffracta　Vain.）である、請求項９又は１０記載の製造方法。 The production method according to claim 9 or 10, wherein the lichen belonging to the genus Sarhogase is Nagasao longissima or Usnea diffracta Vain.
  12. 　抽出したサルオガセ属地衣類抽出物に対して、低温保存処理、加水処理、活性炭処理、ヘキサン洗浄処理及び加熱処理からなる群より選ばれる少なくとも１種の処理により該サルオガセ属地衣類抽出物に含まれるウスニン酸の選択的低減処理を行う、請求項９～１１のいずれか１項記載の製造方法。 Usnic acid contained in the extract of the genus Lithoaceae by at least one treatment selected from the group consisting of low-temperature storage treatment, water treatment, activated carbon treatment, hexane washing treatment, and heat treatment with respect to the extracted Lichenaceae lichen extract The manufacturing method according to any one of claims 9 to 11, wherein the selective reduction treatment is performed.
  13. 　水系溶媒を用いて前記サルオガセ属地衣類を処理し、処理したサルオガセ属地衣類に対して前記アルコール水溶液を用いた抽出を行う、請求項９～１２のいずれか１項記載の製造方法。 The production method according to any one of claims 9 to 12, wherein the Saruogase lichen is treated with an aqueous solvent, and the treated Saruogase lichen is extracted with the aqueous alcohol solution.
  14. 　得られたエンドセリン作用抑制剤中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が３以上である、請求項９～１３のいずれか１項記載の製造方法。 The production method according to any one of claims 9 to 13, wherein the content of difractic acid is 3 or more when the content of usnic acid in the obtained endothelin action inhibitor is 1 on a mass basis.
  15. 　請求項１、３～５及び７のいずれか１項記載の製造方法により得られるサルオガセ属地衣類抽出物を有効成分とする、エンドセリン作用抑制剤。 An endothelin action inhibitor comprising, as an active ingredient, an extract of Ligocephalus lichen obtained by the production method according to any one of claims 1, 3 to 5, and 7.
  16. 　請求項１～１４のいずれか１項記載の製造方法により得られるサルオガセ（Usnea）属地衣類抽出物又はエンドセリン作用抑制剤を有効成分とする、美白剤。 A whitening agent comprising, as an active ingredient, a Usnea garment extract or endothelin action inhibitor obtained by the production method according to any one of claims 1 to 14.
  17. 　サルオガセ（Usnea）属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う抽出物の製造方法であって、前記処理後のウスニン酸の残存率が５０％未満である、ウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。 One or more treatments selected from the group consisting of cryopreservation treatment and hydration treatment are extracted from lichen genus lichen using an aqueous alcohol solution with an alcohol content of more than 70% by volume. The method for producing an extract for carrying out the above-mentioned treatment, wherein the residual rate of usnic acid after the treatment is less than 50%.
  18. 　サルオガセ（Usnea）属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う、ジフラクタ酸高含有かつウスニン酸低含有サルオガセ属地衣類抽出物の製造方法。 One or more treatments selected from the group consisting of cryopreservation treatment and hydration treatment are extracted from lichen genus lichen using an aqueous alcohol solution with an alcohol content of more than 70% by volume. A method for producing a lichenaceae lichen extract with a high content of difractic acid and a low content of usnic acid.
  19. 　前記アルコール水溶液がエタノール水溶液である、請求項１７又は１８記載の製造方法。 The manufacturing method according to claim 17 or 18, wherein the aqueous alcohol solution is an aqueous ethanol solution.
  20. 　前記サルオガセ属地衣類がナガサルオガセ（Usnea　longissima）又はヨコワサルオガセ（Usnea　diffracta　Vain.）である、請求項１７～１９のいずれか１項記載の製造方法。 The manufacturing method according to any one of claims 17 to 19, wherein the lichen belonging to the genus Sargogase is Usa longissima or Usnea diffracta Vain.
  21. 　得られた抽出物中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が２以上である、請求項１８～２０のいずれか１項記載の製造方法。 The production method according to any one of claims 18 to 20, wherein the content of difractic acid is 2 or more when the content of usnic acid in the obtained extract is 1 on a mass basis.
  22. 　サルオガセ（Usnea）属地衣類を、アルコール含有率が７０体積％を超えるアルコール水溶液を用いて抽出し、得られた抽出物に対して低温保存処理及び加水処理からなる群より選ばれる１種以上の処理を行う、エンドセリン作用抑制剤の製造方法。 One or more treatments selected from the group consisting of cryopreservation treatment and hydration treatment are extracted from lichen genus lichen using an aqueous alcohol solution with an alcohol content of more than 70% by volume. A process for producing an endothelin action inhibitor.
  23. 　前記アルコール水溶液がエタノール水溶液である、請求項２２記載の製造方法。 The manufacturing method according to claim 22, wherein the aqueous alcohol solution is an aqueous ethanol solution.
  24. 　前記サルオガセ属地衣類がナガサルオガセ（Usnea　longissima）又はヨコワサルオガセ（Usnea　diffracta　Vain.）である、請求項２２又は２３記載の製造方法。 The manufacturing method according to claim 22 or 23, wherein the lichen belonging to the genus Sarhogase is Nagasao longissima or Usnea diffracta Vain.
  25. 　得られたエンドセリン作用抑制剤中のウスニン酸含有量を質量基準で１とした場合、ジフラクタ酸含有量が２以上である、請求項２２～２４のいずれか１項記載の製造方法。 The production method according to any one of claims 22 to 24, wherein the content of difractic acid is 2 or more when the content of usnic acid in the obtained endothelin action inhibitor is 1 on a mass basis.
  26. 　請求項１７～２５のいずれか１項記載の製造方法により得られるサルオガセ（Usnea）属地衣類抽出物又はエンドセリン作用抑制剤を有効成分とする、美白剤。
     
     
          A whitening agent comprising, as an active ingredient, a Usnea garment extract obtained by the production method according to any one of claims 17 to 25 or an endothelin action inhibitor.


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CN106749137A (en) * 2016-11-30 2017-05-31 宝鸡文理学院 A kind of method that usnic acid is extracted from stellate cladonia fruticose thallus
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1089089A (en) * 1993-01-05 1994-07-13 殷振鹏 The method for making of heart stablizing and blood pressure-lowering health medicated tea
CN1119952A (en) * 1995-08-12 1996-04-10 张毓骅 External applying medicine powder for treating essential hypertension
JP2000034233A (en) * 1998-07-15 2000-02-02 Sumitomo Forestry Co Ltd Inhibitor of production of nitric oxide
JP2012006853A (en) * 2010-06-23 2012-01-12 Kansai Univ Melanin formation inhibiting agent, cosmetic agent, skin preparation for external use, quasi-drug, and anticancer drug

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010150173A (en) * 2008-12-25 2010-07-08 Kose Corp Skin whitening agent, melanogenesis inhibitor, and skin whitening skin external preparation
CN101664401A (en) * 2009-08-25 2010-03-10 华东师范大学 Application of usnic acid in preparation of drugs inhibiting angiogenesis

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1089089A (en) * 1993-01-05 1994-07-13 殷振鹏 The method for making of heart stablizing and blood pressure-lowering health medicated tea
CN1119952A (en) * 1995-08-12 1996-04-10 张毓骅 External applying medicine powder for treating essential hypertension
JP2000034233A (en) * 1998-07-15 2000-02-02 Sumitomo Forestry Co Ltd Inhibitor of production of nitric oxide
JP2012006853A (en) * 2010-06-23 2012-01-12 Kansai Univ Melanin formation inhibiting agent, cosmetic agent, skin preparation for external use, quasi-drug, and anticancer drug

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
"reduced-size edition 30th print", KAGAKU DAIJITEN 1, 1987, pages 781 - 782 *
"reduced-size edition 30th print", KAGAKU DAIJITEN 4, 1987, pages 457 *
BATUBARA I. ET AL.: "Potency of Indonesian Medicinal Plants as Tyrosinase Inhibitor and Antioxidant Agent", JOURNAL OF BIOLOGICAL SCIENCES, vol. 10, no. 2, 2010, pages 138 - 144 *
CHUYAKU DAIJITEN 3RD EDITION, vol. 2, no. 3RD ED, 1 May 1998 (1998-05-01), pages 1274 - 1275 *
FRANKOS VH.: "NTP NOMINATION FOR USNIC ACID AND USNEA BARBATA HERB", NATIONAL TOXICOLOGY PROGRAM, 2005, Retrieved from the Internet <URL:http://ntp.niehs.nih.gov/ntp/htdocs/ChemBackground/ExSumPdf/UsnicAcid508.pdf> *
ISAO YOSHIMURA ET AL.: "Secondary metabolic substances of Usnea diffracta Vain", BULLETIN OF KOCHI GAKUEN COLLEGE, vol. 23, 1992, pages 687 - 697 *
MAHADIK ND ET AL.: "Cardiovascular-protective, Antioxidative, and Antimicrobial Properties of Natural Thallus of Lichen Usnea complanata", LATIN AMERICAN JOURNAL OF PHARMACY, vol. 30, no. 2, 2011, pages 220 - 228 *
OKUYAMA E. ET AL.: "Usnic acid and diffractaic acid as analgesic and antipyretic components of Usnea diffracta.", PLANTA MED., vol. 61, no. 2, 1995, pages 113 - 115 *
RAWAT MSM. ET AL.: "Chemical study on Garhwal Himalayan Lichen: Usnea emidotteries", INDIAN JOURNAL OF CHEMISTRY, vol. 45B, no. 11, 2006, pages 2566 - 2570 *

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