JP2006036731A - Bamboo grass extract and its use - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a preparation method for efficiently obtaining a bamboo grass extract, taking into account the extracted volume of a compound which has not been focused as an ingredient of the bamboo grass extract, and to provide the bamboo grass extract excellent in a wide range of pharmacological actions, especially in antibacterial and antimutagenic properties. <P>SOLUTION: The bamboo grass extract contains ≥0.5 mg/g each of phenyl propanoid, acetic acid, formic acid and 3-hydroxypyridine and is obtained by cooking bamboo leaves and/or culms and subsequently subjecting them to extraction with an aqueous solvent. The sugar content in the bamboo grass extract is ≤50 mg/g when heating residue is adjusted to 40-60 wt.%. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to Sasa extract useful as an antibacterial agent, antiviral agent, antitumor agent, antimutagenic agent, anti-inflammatory agent, and use of the extract. The Sasa extract of the present invention can efficiently produce specific components by steaming persimmon leaves and / or persimmons with steam having improved dryness.

  Sasa has long been recognized for its medicinal properties, and Sasa extract has also been reported to have many pharmacological effects such as anti-inflammatory / anti-ulcer, antihypertensive, and antitumor (J. Hokkaido For. Prod. Res. Inst.Vol.9, No.6,1995, Showa Medical Society Journal Volume 48, No. 5, 595-600, 1988, etc.).

As an active ingredient in Sasa extract, the antitumor property of soluble hemicellulose has attracted attention, and among them, polysaccharides centered on xylooligosaccharide are considered to play an important role in the expression of antitumor effect. JP-A-06-197800, JP-A-11-199502 and the like disclose methods for efficiently extracting saccharides by subjecting leaves and / or straws to high temperature and high pressure saturated steam treatment. Japanese Patent Application Laid-Open No. 2002-322079 uses a Sasa extract obtained by high-temperature and high-pressure saturated steam treatment as an antibacterial and antibacterial agent against Helicobacter pylori.
None of the above-mentioned known documents mentions any components other than sugars, and the steam to be cooked does not go through means for improving the dryness, resulting in a difference from the extract of the present invention. It is likely that it was. Further, if the saccharide content is higher than necessary, the growth of fungi is promoted and it is not suitable as an antibacterial agent.
Japanese Unexamined Patent Publication No. 06-197800 Japanese Unexamined Patent Publication No. 11-199502 JP 2002-322079 A J. Hokkaido For. Prod. Res. Inst. Vol. 9, No. 6, 1995 Showa Medical Society Volume 48, No. 5, 595-600, 1988

  The object of the present invention is to provide a production method capable of efficiently obtaining compounds in consideration of the extraction amount of compounds that have not been noticed as components of Sasa extract so far, and thus having a wide range of pharmacological actions, particularly antibacterial properties. The object is to provide a Sasa extract excellent in antimutagenicity.

  The present invention contains 0.5 mg / g or more of phenylpropanoid, acetic acid, formic acid, and 3-hydroxypyridine, each of which is obtained by steaming bamboo leaves and / or straw and then extracting with an aqueous solvent. It is related with the Sasa extract characterized by the content of saccharides when 50-40 weight% is prepared.

  The present invention also relates to the above Sasa extract, wherein the phenylpropanoid is at least one compound selected from coumaric acid, ferulic acid, caffeic acid, and vanillin.

  Moreover, this invention relates to the said Sasa extract whose content of formic acid is 10 mg / g or more.

  Moreover, this invention relates to the said Sasa extract whose content of 3-hydroxypyridine is 1 mg / g or more.

  The present invention also provides that the water content of the bamboo leaf and / or koji before cooking is not more than 10% by weight with respect to the whole koji leaf and / or koji, and the steam to be cooked is a pressure reducing valve and gas-liquid. It is related with the said Sasa extract which is the dry steam which improved the dryness by the separator.

  Moreover, this invention relates to the said Sasa extract characterized by steaming temperature being 160-190 degreeC, and aqueous-solvent extraction temperature being 100-150 degreeC.

  Moreover, this invention relates to the said Sasa extract whose aqueous solvent is a hot water or alkaline aqueous solution.

  Further, the present invention is to extract persimmon leaves and / or persimmons from 160 to 190 ° C. for 30 seconds to 1 hour with dry steam whose dryness is improved by a pressure reducing valve and a gas-liquid separator, and then extract with an aqueous solvent. The present invention relates to a method for producing Sasa extract.

  Moreover, this invention relates to the said Sasa extract manufacturing method further including the process of expelling air from a cooking apparatus before and / or during cooking.

  Moreover, this invention relates to the antibacterial agent containing the said Sasa extract.

  Moreover, this invention relates to the antimutagenic agent containing the said Sasa extract.

  By this invention, the Sasa extract excellent in antibacterial property and antimutagenicity is obtained.

The spider used in the present invention is preferably one belonging to the genus Kumazasa, for example, Gotenbazasa, Katsuragazasa, Suzutake, Miyama Kumazasa, Negararidake, Miyakozasa, Hosobazasa, Kumizasa, Kumazasa, Chugokusasa, etc. Among them, Kumazasa is most preferable. In addition, it is well known that the components accumulated in the leaves and persimmon fluctuate depending on the season, and the persimmons harvested from July to September are optimal for use in the present invention.
The dried bamboo leaves and / or straw used in the steaming of the present invention are preferably dried, and the preferred moisture is 10% or less with respect to the whole bamboo leaves and / or straw, and if it is more than that, spoilage, fermentation There is a risk that the component will be altered by. About the form of the bamboo leaf and / or the bamboo shoot used for the steaming of the present invention, those which have been cut into pieces are preferable, and the preferred size is 0.5 to 20 mm. If it is larger than 20 mm, the extraction efficiency is deteriorated.

  The steaming of the present invention can be carried out by any device that can withstand high temperatures and high pressures, and the temperature during cooking is most preferably 160 to 190 ° C. If the temperature is lower than 160 ° C, the cell wall is hardly destroyed by the decomposition of hemicellulose in the cocoon, so that the desired component cannot be sufficiently extracted. is there. The cooking time is preferably between 30 seconds and 1 hour, more preferably between 5 minutes and 20 minutes. If the time is shorter than 30 seconds, the hydrolysis of hemicellulose becomes insufficient and the amount of components decreases, and it is longer than 1 hour. Then, decomposition of the obtained component occurs and the amount of the obtained component is reduced. When releasing the pressurization after completion of cooking, it is preferable to open the apparatus at once to bring the pressure to atmospheric pressure, since the destruction of the tissue of the sasa leaves or straw will progress further and the extraction efficiency will improve.

  The steam used for steaming is most preferably steam of an apparatus provided with a steam / water separator and a pressure reducing valve in the middle of a steam pipe connecting a steam generating source such as a boiler. By using the apparatus, the dryness of the steam is improved. As the degree of dryness is high and the amount of condensate of steam called drain is small, it is possible to efficiently extract a specific component and appropriately control the extraction amount of sugars. Since there is no simple measurement method for the dryness of the vapor, it is difficult to directly measure it with an ordinary manufacturing apparatus. Therefore, in order to obtain dry steam, it is preferable to use a gas-liquid separator that has been confirmed to be able to achieve a dryness of 99% or more by a dryness measuring system owned by a gas-liquid separator manufacturer or the like. There are a centrifugal type and a baffle type as the gas-liquid separator, but the baffle type is preferable because it can stably achieve a high dryness regardless of the amount of steam.

  If air remains in the steaming apparatus during steaming, contact between saturated steam and Sasa leaves or straw will be hindered, resulting in insufficient steaming and reduced extraction efficiency. As a means for avoiding the remaining of the air, it is only necessary to introduce steam with the manual or automatic air vent valve opened and to expel the air from the cooking apparatus. The vapor temperature for expelling air may be 80 to 200 ° C., preferably 100 to 180 ° C., and the time for expelling air is 0.5 to 60 minutes.

  The extraction method using an aqueous solvent is not particularly limited, but after cooking, the aqueous solvent is added while cooling or maintaining the temperature, and steamed Sasa leaves or steamed leaves are added to the aqueous solvent under conditions of room temperature to 180 ° C. Soak the salmon.

  The aqueous solvent used in the present invention is preferably a water-soluble solvent such as methanol, ethanol, acetone or the like, or an aqueous solution thereof, or water. If permitted by the use of the Sasa extract, the extraction efficiency can be increased by using an alkaline aqueous solution. . The alkaline aqueous solution used in the present invention is preferably sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogen carbonate, ammonium hydrogen carbonate or the like.

  In the present invention, the steaming step and the extraction step may be repeatedly performed without departing from the gist of the present invention. In addition, another process such as a holding process may be provided before and after each process.

  Although the mechanism by which phenylpropanoid, acetic acid, formic acid, and 3-hydroxypyridine contained in the extract of the present invention exhibit antibacterial properties and antimutagenic properties has not been clarified, each component can be used alone due to the synergistic effect of each component. It is presumed to exert a greater effect than the component.

  Examples of the phenylpropanoids contained in the Sasa extract of the present invention include coumaric acid, ferulic acid, caffeic acid, vanillin, etc., and the content of coumaric acid is the largest, followed by ferulic acid. The amount of phenylpropanoid necessary to effectively act as an antibacterial agent or antimutagenic agent is 0.5 mg / g or more in the Sasa extract as the total amount of all phenylpropanoids, preferably If coumaric acid alone is 1 mg / g or more, high antibacterial properties and antimutagenic properties can be obtained.

  The content of acetic acid and formic acid contained in the Sasa extract of the present invention is required to be 0.5 mg / g or more, preferably 5 mg / g or more formic acid alone, more preferably 10 mg / g or more formic acid alone. is there.

  The detailed synthesis route of 3-hydroxypyridine contained in the Sasa extract of the present invention is unknown, but it is presumed to be a substance produced when steaming bamboo leaves and / or straw, and it is an antibacterial agent or antimutation. The content necessary as a chemical agent is 0.5 mg / g or more, preferably 1 mg / g or more, more preferably 2 mg / g or more.

  The saccharide contained in the Sasa extract of the present invention is a monosaccharide and / or polysaccharide composed of xylose, arabinose, glucose, mannose, galactose, etc., and is preferably contained when the heating residue is adjusted to 40 to 60% by weight. When the amount is 50 mg / g or less and more, the pharmacological actions such as antibacterial, antiviral, antitumor, antimutagenic, and anti-inflammatory properties which are the effects of the extract of the present invention are impaired. In particular, an excessive amount of saccharides promotes the growth of fungi, and those in which the fungus has grown are not suitable for normal use as an extract. The heating residue referred to in the present invention is a so-called solid content, and means the content of the residue obtained by removing low-boiling substances such as water by heating. For example, the amount left by heating in a 100 ° C. oven for 60 minutes Is divided by the amount before heating.

  Phenylpropanoid, acetic acid, formic acid and 3-hydroxypyridine can be quantified by liquid chromatography, gas chromatography or the like. Saccharides can be quantified by ion chromatography.

  The form of the Sasa extract of the present invention may be a solution or a solid, and may be a mixture with other compounds. In the case of a mixture, it may be processed by spray drying, freeze drying, addition of a shaping agent such as dextrin. Further, it may be subjected to a selection process such as filtration, column purification, and solvent washing.

When the Sasa extract of the present invention is used as an antibacterial agent, it exhibits an antibacterial action against various fungi, such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, tetanus, Clostridium gas gangrene, Candida Fungus, ringworm fungus, mold fungus, spider fungus, Aspergillus spp., Cryptococcus spp., Coccidioides spp., Histoplasma spp. Anaerobic gram-positive cocci such as Steiptococus constellatus, Atopobium parvulum, Gemella morbillorum,
Anaerobic Gram gonococci such as Propionibacterium acnes, Propionibacterium granulosum, Eggerthella lenta, Actinomyces odontolyticus, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium pseudolongum, Mobiluncus spp.
Bacteroides fragilis penicillin resistance, Bacteroides fragilis carbapenem resistance, Bacteroides vulgatus, Bacteroides distasonis, Bacteroides ovatus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, Bacteroides ureot, Preactella intermedia, Prevoterlla melaninogenica, Prevotella oralis, Prevotella oris, Porphyromonas asaccharolytica, Porphyromonas gingivalis, Fusobacterium nucleatum, Fusobacterium varium, Fusobacterium necrophrum, Bilophila wadsworthia, Desulfovibrio piger, Capnocytophage

Clostridium diffioile, Clostriduim sordellii, Clostridium septicum, Clostridium perfringens, Clostridium ramosum, Clostridium clostridiiforme, Clsotridium bifermentans, Clostridium sordellii, Clostridium novyi Type A, Clostridium sporogenbotulinum, Clostridium sporiibot
Facultative anaerobic gram-negative bacilli such as Escherichia coli, Enterobacter cloacae, Flavobacterium meningosepticum, Pseudeomonas aeruginosa,
Facultative anaerobic Gram-positive cocci such as Staphylococcus aureus MSSA, Staphylococcus aureus MRSA, Staphylococcus epidermidis, Staphylococcus hemolyticus, Gardnerella vaginalis,
It exhibits antibacterial properties against bacteria such as Lactobacillus acidophilus, Lactobacillus brevis ss. Brevis, Lactobacillus casei ss. Casei, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus salivarius ss. Salivarius.

  The usage form of the antibacterial agent or antimutagenic agent of the present invention is not particularly limited, and may be any of solid, liquid, and semisolid forms, and can be oral or parenteral. Ingestion can be made by adding to food as a food additive. When used as a food additive, the amount added is not particularly limited, and may be appropriately determined according to the type of food. For example, liquid foods such as soft drinks and carbonated drinks, and solid foods such as confectionery and other various foods can be added and used. In addition, an example of an administration method when the antibacterial agent or antimutagenic agent of the present invention is administered to the human body is as follows. Administration can be performed by various methods, for example, oral administration using tablets, capsules, granules, syrups and the like. The oral preparation can be produced according to a normal production method. For example, excipients such as starch, lactose and mannitol, binders such as sodium carboxymethylcellulose and hydroxypropylcellulose, disintegrants such as crystalline cellulose and carboxymethylcellulose calcium, lubricants such as talc and magnesium stearate, light anhydrous It can be produced as a tablet, capsule, granule, etc. by appropriately combining and formulating a fluidity improver such as silicic acid.

  It can also be used effectively by adding it to cosmetics and applying it to the skin. When added to cosmetics, the addition amount may be appropriately determined within a range that does not impair the original function of the cosmetics.

  Other uses of the antibacterial agent of the present invention include sprays, suppositories, preservatives, brushing agents, soaps, shampoos, lotions, ointments, skin adhesive films, disinfectants, bath preparations, cosmetics, and other medical products. Or medical aids, earplugs, gloves, hats, lab coats, eyebands and other medical supplies, air purifiers, coolers, vacuum cleaners, ventilation fans, masks and other filters, carpets, floorboards, wallboards, wallpaper, etc. Protective sheets such as equipment, furniture and sundries, protective covers, protective plates, masterbatches, paints, inks, clothes, skin guards, bedding, bandages, gauze, handkerchiefs, pennants, cloths, daily cloth products, wrapping paper, Examples include daily paper products such as notebooks, memos, and wet tissues.

Hereinafter, the present invention will be described in more detail with reference to examples. In addition,% shows weight%.
<Production example of Sasa extract>
2 kg of dried Kumizasa leaves and straws crushed to 0.5-5 mm and charged in a high-pressure steam can were charged with 2 MPa of high-pressure steam generated by a once-through boiler, with a dryness measuring system of 99. Gas-liquid separation was performed with a separator that was confirmed to be achieved up to 5%, and steam with improved dryness by reducing the pressure to 1.6 MPa with a pressure reducing valve was introduced into a high-pressure steam can. At the beginning of steam introduction, the air vent valve is opened and the steam is allowed to pass through the steaming can for 10 minutes at 100 ° C. to expel air, the air vent valve is closed and the temperature is raised from 100 ° C. to 180 ° C. over 10 minutes. The pressure was released immediately after holding for 10 minutes. Leave high pressure steamed leaves and straw in a pressure extractor, add 8 kg of water, raise the temperature from room temperature to 110 ° C. over 30 minutes, hold at that temperature for 5 minutes, and then release the pressurized state over 1 hour . The extract was filtered and concentrated under reduced pressure to obtain a Sasa extract with a heating residue of 50%.

(Comparative Example 1)
Sasa extract was produced with reference to the method described in Example 1 of JP-A-06-197800. That is, 20 kg of water was added to 2 kg of dried Kumizasa leaves and straw that had been crushed to 0.5 to 5 mm, and filtered for 3 hours at 80 ° C., followed by filtration. This extraction operation was repeated twice to prepare Kumizasa from which the hot water extract was removed. Next, this was adjusted to a moisture content of 45%, charged into a high-pressure steam can, and the 2 MPa high-pressure steam generated in the once-through boiler was decompressed to 1.6 MPa by a pressure reducing valve and directly introduced into the high-pressure steam can. The temperature was raised from room temperature to 180 ° C. over 10 minutes, held at that temperature for 10 minutes, and then the pressure was gradually released. High pressure steamed leaves and straw were charged into a pressure extraction can, 8 kg of water was added, the temperature was raised from room temperature to 110 ° C. over 30 minutes, held at that temperature for 5 minutes, and then the pressurized state was released over 1 hour. . The extract was filtered and concentrated under reduced pressure to obtain a Sasa extract with a heating residue of 50%.

(Comparative Example 2)
Sasa extract was produced with reference to the method described in Example 1 of JP-A-11-199502. That is, 2 kg of dried Kumizasa leaves and straw crushed to 0.5-5 mm and put in a heat-resistant bag, charged in a pressure extraction can, added 37 kg of water, and raised from room temperature to 120 ° C. over 30 minutes. It was warmed and held at that temperature for 10 minutes, and then the pressurized state was released over 1 hour. Next, the hot water in the pressure extraction can was cooled to 80 ° C. with cooling water, and then the lid of the pressure extraction can was opened, and the leaves and straw were taken out and placed in a screw press to adjust the water content to 50%. The extract separated in this step is added to the hot water in the previous pressure extraction can. Next, the solid content adjusted to a moisture content of 50% is charged into a high-pressure steam can, and the 2 MPa high-pressure steam generated by the once-through boiler is reduced to 1.6 MPa by a pressure reducing valve and directly introduced into the high-pressure steam can. did. The temperature was raised from room temperature to 180 ° C. over 10 minutes, held at that temperature for 10 minutes, and then the pressure was gradually released. The high-pressure steamed leaves and straw were charged again in a pressure extraction can and heated from room temperature to 110 ° C. over 30 minutes, held at that temperature for 5 minutes, and then the pressurized state was released over 1 hour. The extract was filtered and concentrated under reduced pressure to obtain a Sasa extract with a heating residue of 50%.

<Evaluation of Sasa extract>
(Composition analysis method)
Qualitative and quantitative analysis of acetic acid, formic acid, coumaric acid, ferulic acid, and 3-hydroxypyridine was performed using a gas chromatograph mass spectrometer equipped with a J & W innowax 30 m × 0.25 mm, 0.25 um column.

  In addition, using DXEX-500 sugar analyzer manufactured by Dionex equipped with CarboPac PA1 as column and electrochemical detector as detector, arabinose, rhamnose, galactose, glucose, xylose, mannose, fructose, ribose, xylobiose, xylotriose, xylotetra Qualitative and quantitative analysis of ose, xylopentaose, xylohexaose, arabinobiose, arabinotriose, arabinotetraose, arabinopentaose, arabinohexaose and the total value of all components as sugar content did.

(Antimicrobial test method)
The test bacteria are as follows.
Escherichia coli NBRC 3962
Staphylococcus aureus NBRC12732
Pseudomonas aeruginosa: Pseudomonas aeruginosa NBRC13275
The minimum drug concentration (MIC) at which the Sasa extract diluted at various magnifications was brought into contact with Müller Hinton broth with a substantially constant number of bacteria and the growth of the bacteria was inhibited after a certain period of time was determined and used as an index of antibacterial effect.
The smaller the MIC, the higher the antibacterial effect.

(Anti-mutagenicity test method)
The number of revertant colonies was counted using Salmonella TA100 as the strain and B (a) P; benzo (a) pyrene as the mutagen. As an index of antimutagenicity, a relative value obtained by converting the number of revertant colonies not added with the Sasa extract sample to 100 is shown. The smaller the value, the higher the antimutagenicity.

(Evaluation results)
Table 1 shows the evaluation results of the Sasa extract obtained in Production Example, Comparative Example 1, and Comparative Example 2 by the above evaluation method.

（注１）ササ抽出液未添加を１００とした場合の相対値
（注２）ササ抽出物製造後速やかに菌が繁殖したと考えられエキスとしては全く不適当 であることを示している。

(Note 1) Relative value when Sasa extract is not added as 100 (Note 2) This indicates that the fungus has proliferated quickly after the Sasa extract is produced, indicating that it is totally inappropriate as an extract.

Claims (10)

Boiled persimmon leaves and / or persimmons, extracted with an aqueous solvent, each containing 0.5 mg / g or more of phenylpropanoid, acetic acid, formic acid, 3-hydroxypyridine, and a heating residue of 40 to Sasa extract characterized by having a saccharide content of 50 mg / g or less when adjusted to 60% by weight. The Sasa extract according to claim 1, wherein the phenylpropanoid is at least one compound selected from coumaric acid, ferulic acid, caffeic acid, and vanillin. The Sasa extract according to claim 1 or 2, wherein the formic acid content is 5 mg / g or more. The Sasa extract according to any one of claims 1 to 3, wherein the content of 3-hydroxypyridine is 1 mg / g or more. The moisture of the bamboo leaf and / or the bamboo shoot before cooking is 10% or less by weight with respect to the whole bamboo leaf and / or the bamboo shoot, and the steam to be cooked is dried by a pressure reducing valve and a gas-liquid separator. The Sasa extract according to any one of claims 1 to 4, which is an improved dry steam. Steaming temperature is 160-190 degreeC, and aqueous-solvent extraction temperature is 100-150 degreeC, The Sasa extract in any one of Claims 1-5 characterized by the above-mentioned. The aqueous extract is hot water or an alkaline aqueous solution. Bamboo leaves and / or persimmons are steamed at 160-190 ° C. for 30 seconds to 1 hour with dry steam whose dryness is improved by a pressure reducing valve and a gas-liquid separator, and then extracted with an aqueous solvent. Sasa extract manufacturing method. Furthermore, the method for manufacturing Sasa extract according to claim 8, further comprising the step of expelling air from the steaming apparatus before and / or during steaming.
The antibacterial agent containing the Sasa extract in any one of Claims 1-7.