JP2007277120A - Method for extracting ganoderma lucidum extract and bleaching agent containing the ganoderma lucidum extract - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To improve extraction efficiency in a method for extracting Ganoderma lucidum with a super critical fluid without reducing melanin suppression action which the Ganoderma lucidum extract has, and apply the Ganoderma lucidum extract obtained by the method to skin care preparations for external use and the like as a bleaching agent. <P>SOLUTION: In the method for preparing the Ganoderma lucidum extract having melanin production suppressing action from Ganoderma lucidum, Ganoderma lucidum is extracted with the fluid in a state of 30-150°C temperature range and 42-100 MPa pressure range. The bleaching agent contains the Ganoderma lucidum extract prepared by the method. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  TECHNICAL FIELD The present invention relates to a method for extracting a nanentake extract having an inhibitory action on melanin production from nanentake, and a whitening agent containing the nanentake extract extracted by the method.

  Skin spots such as sunburn, freckles, and liver spots occur with aging and are difficult to increase or disappear with aging. The mechanism of the development of these pigmentations has not yet been clarified, but melanin pigments are produced and produced in melanin-producing granules called melanosomes in melanocytes existing in epidermal cells by sun rays (especially ultraviolet rays). It is thought to be caused by the diffusion of melanin pigments to adjacent cells.

  Development of a substance capable of restoring such pigmentation to normal skin color is strongly desired, and many substances have been developed so far. For example, L-ascorbic acid, kojic acid, arbutin and the like are known.

  The present applicants have been developing whitening agents for a long time. For example, whitening agents composed of ergosterol derivatives (see, for example, Patent Document 1), ganoderol B (see, Patent Document 2), and the like, kakukaku reishi extract And a melanin production inhibitor containing a microcirculation improving agent (see Patent Document 3). In particular, the Kazuno Ganoderma extract is not toxic, has an excellent whitening effect, and is a useful extract as a whitening agent derived from natural products.

  Recently, with respect to mushrooms such as Kakuno Reishi, the mushroom extract extracted with carbon dioxide in a temperature range of 20 to 70 ° C. and a pressure range of 7 to 40 MPa is superior in the melanin production inhibitory effect compared to the extract with ethanol. (See Patent Document 4).

Japanese Patent Application Laid-Open No. 2002-114685 JP 2002-284690 A JP 2003-171254 A JP 2005-104873 A

  The supercritical carbon dioxide extraction method has been attracting attention for reasons such as simple extraction process, non-irritating and harmless, and practically inactive, so that there is little alteration due to chemical changes in the extract. However, there is a problem that the extraction efficiency is not good as compared with the extraction method using alcohol or the like, and the extraction efficiency of mannentake extract obtained by extracting mannenambo with supercritical carbon dioxide was low.

  The present invention has been made in view of the above circumstances, and the object thereof is a method for extracting mannentake extract having a melanin production inhibitory action with a supercritical fluid, and the extraction of mannentake extract without reducing the melanin production inhibitory action. An object of the present invention is to provide an extraction method capable of improving the efficiency, and to provide a whitening agent containing an extract of Mannentake extracted by the method.

  In order to solve the above-mentioned problems, the present inventors have conducted research on extraction conditions in the supercritical fluid extraction method, and as a result, the supercritical fluid has a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa. Alternatively, extraction with a fluid in the vicinity thereof dramatically improves the extraction efficiency of the garlic mushroom extract, and the extract has a melanin production inhibitory action superior to that of the alcohol extract, It has been found that it has the same or higher melanin production inhibitory action and higher safety than the extract of garlic mushroom extracted by the fluid at or near criticality, and has completed the present invention.

  That is, the first invention is a method for extracting a garlic mushroom extract having an inhibitory action on melanin production from garlic bamboo, wherein the garlic mushroom is extracted with a fluid in a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa. It is an extraction method of the nanentake extract which has the said melanin production inhibitory effect characterized by the above-mentioned.

  In addition, the second invention is a method for extracting a kakukaku reishi extract having a melanin production inhibitory action from kakukaku reishi, which is in a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa. Extracting with the fluid which exists in the above, It is the extraction method of the deer antrum extract which has the said melanin production inhibitory effect.

  Further, the third invention is a method for extracting a nanentake extract having an inhibitory action on melanin production from banyan bamboo, wherein the banyan bamboo is extracted with carbon dioxide in a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa. It is an extraction method of the nanentake extract which has the said melanin production inhibitory action characterized by doing.

  The fourth invention is a method for extracting a kakukaku reishi extract having a melanin production inhibitory action from kakukaku reishi, wherein the koku kaku reishi is in a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa. The method for extracting a deer anteater extract having an inhibitory action on melanin production, characterized by being extracted with carbon dioxide.

  Furthermore, 5th invention is a whitening agent containing the nanentake extract extracted by one of the said methods.

  Moreover, 6th invention is the whitening agent containing the kakukaku reishi extract extracted by the said method.

  ADVANTAGE OF THE INVENTION According to this invention, the highly safe mannentake extract which has the outstanding inhibitory effect of melanin production is obtained efficiently and with a high yield by the supercritical fluid extraction method under specific conditions. The Mannentake extract is applied as a whitening agent in various fields such as cosmetics including quasi-drugs, pharmaceuticals, and foods, and is used in the form of cosmetics, pharmaceutical compositions, foods, and the like. In particular, it is preferably used in the form of a skin external preparation externally applied to the skin by an administration method in which it is directly applied to or spread on the skin and exhibits an excellent whitening effect.

  Hereinafter, the best mode for carrying out the present invention will be described in detail.

  Ganoderma mushrooms are basidiomycetous mushrooms and are classified into the genus Mushrooms (referred to by the school as the genus Mushrooms). There are red ganoderma (red turf or simply ganoderma, scientific name: Ganoderma lucidum) and black ganoderma (black turf, purple turf, scientific name: Ganoderma japonicum, Ganoderma sinense, Ganoderma atrum). In the present invention, it is preferable to use these, and it is particularly preferable to use Kakukaku Ganoderma (Kanokakushiba, scientific name: often described as Ganoderma aboinense), which is a kind of Ganoderma (Akashiba). Further, other mushrooms belonging to the genus Amanita may be used. In addition, Aoshiba, Akashiba, Koshiba, Shiroshiba, Kuroshiba and Shishiba are classified and listed as effective ganoderma in Chinese old drug books “Shinnohonkusa” and “Honso Tuname”. May be used. Moreover, as for the above-mentioned mannambose, a natural native product may be used and a cultivated product may be used.

  Examples of the part of the bamboo shoot used for extraction include fruit bodies, mycelia, mycelium cultures, spores, and the like, preferably fruit bodies. In the extraction, these cells may be the cells themselves or those obtained by drying, pulverizing or chopping the cells.

  In the present invention, the bamboo shoot is extracted by a fluid in a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa. When the fluid is in this range, an extract having excellent extraction efficiency, low toxicity, and high effectiveness can be obtained. A more preferable fluid is a fluid in a temperature range of 30 to 90 ° C. and a pressure range of 42 to 80 MPa.

  In the above range, the fluid is supercritical or in the vicinity thereof.

  Examples of the fluid include carbon dioxide, water, methanol, and Freon. Of these, carbon dioxide is preferred. Carbon dioxide becomes supercritical carbon dioxide at a temperature and pressure above the critical point (temperature 30.9 ° C., pressure 7.38 MPa). In the present invention, the temperature range is 30.9 to 150 ° C. and the pressure range is 42 to 100 MPa. It is preferable to use supercritical carbon dioxide as described above. More preferred is supercritical carbon dioxide in a temperature range of 30.9 to 90 ° C. and a pressure range of 42 to 80 MPa.

  As for the pressure range, there is a critical point at the lower limit, and the critical pressure is important. However, as will be clarified in the examples of the extract to be described later about the extraction efficiency, it is also from a practical point of view. More preferably, it is 45-60 MPa. Further, the extraction time and the fluid supply amount are not particularly limited, and may be performed within an arbitrary range.

  In the extraction, a cosolvent (entrainer) can be used. Examples of the co-solvent include water, ethanol, acetone and the like. Among these, water and ethanol are preferable from the viewpoint of safety.

  The extraction method is not particularly limited except for the temperature and pressure, and is performed by a general method. For example, in an extraction tank, a fluid such as carbon dioxide under the above extraction conditions is continuously added to mannentake. This can be done by blowing. After the extraction, a fluid such as carbon dioxide containing an extract of garlic mushroom is introduced into a separation tank, and separated by a generally used method, for example, a method of lowering pressure or temperature. At this time, the separation tank can be filled with a solvent and separated as a solution of the extract of mannentake.

  When separated as the solution, it may be used as it is, but if necessary, after filtration, treatment such as concentration, dilution, deodorization, and drying may be appropriately performed.

  The garlic mushroom extract extracted by the above method according to the present invention (hereinafter simply referred to as mannentum mushroom extract) has an excellent melanin production inhibitory action. Therefore, the garlic mushroom extract according to the present invention is useful as a whitening agent (hereinafter referred to as whitening agent (component)) as a component.

  The whitening agent (component) has a very wide application range, and is applied to various fields such as cosmetics, pharmaceutics, foods including quasi-drugs, and preferably used by being blended into a skin external preparation.

  In the present invention, a whitening agent is prepared by containing mannentake extract as a component having a whitening effect, and the whitening agent exhibits an excellent whitening effect.

  In the present invention, when preparing the whitening agent, the amount of the extract of the white mushroom may be 100% by mass in the total amount of the whitening agent, but an amount effective for exerting the whitening effect is sufficient. From the viewpoint of ease of preparation and use of the whitening agent, and from the point of practicality, it is preferably an effective amount in the range of 0.0001 to 10.0% by mass in the total amount of the whitening agent. More preferably, it is 0.01-5.0 mass%.

  Therefore, in the whitening agent according to the present invention, “contains” regarding the content is a concept including “consisting of”.

  The whitening agent in the present invention is usually used in cosmetics, pharmaceutical compositions, foods and the like including quasi-drugs, as long as the effect of the present invention is not impaired, except for containing 100% by mass of Mannentake extract. Other components can be blended as appropriate. Examples of the other components include oils, surfactants, powders, thickeners, moisturizers, ultraviolet absorbers, antioxidants, antiseptics, anti-inflammatory agents, plant extracts, and whitening agents other than those described above (components) ), Flavor, water, excipient, stabilizer, wetting agent, emulsifier, absorption enhancer, pH adjuster, diluent, carrier, dissolution aid, flavoring agent, preservative, colorant, coating agent, sweetener And sour agents.

  The whitening agent includes, for example, cosmetics including quasi drugs, pharmaceutical compositions (internal use, external use), nutritional functional foods, foods for specified health use in various fields such as cosmetics including quasi drugs, pharmaceuticals, and foods. It can be used in the form of foods such as beverages and general foods, and can take various dosage forms suitable for each.

  Specifically, for example, lotion, cosmetic oil, milky lotion, cream, gel, essence (beauty serum), pack mask, white powder / dusting, makeup base, foundation, sunscreen cream, sunscreen lotion, suntan oil , Capsules, granules, pills, powders, emulsions, troches, liquids for internal use, syrups, ointments, tinctures, spirits, poultices, patches, aerosols, emulsions, lotions, suspensions , Solution, emulsion, oral, injection, tablet, suppository, beverage (alcoholic beverage, carbonated beverage, fruit juice beverage, vitamin beverage, nutritional beverage, tea beverage, mineral beverage, lactic acid bacteria beverage, milk beverage, sports beverage , Soft drinks, etc.), cookies, gummi, candy, caramel, chocolate, jelly, tablet confectionery, noodles, bread, processed rice products, seasonings (sauce, soy sauce, etc.), gum, candy Confectionery, solution system, solubilization system, emulsification system, powder system, powder dispersion system, oil liquid system, gel system, gel system, ointment system, aerosol system, mousse system, water-oil two-layer system, water-oil -Product forms and dosage forms such as powder three-layer systems can be mentioned.

The usage and dosage of the whitening agent in the case of the pharmaceutical composition are not particularly limited and are appropriately determined depending on individual differences such as symptoms, age, body weight, etc., but in the case of oral administration, usually 0.01-100 mg / Kg, preferably 0.1 to 10 mg / kg, and this amount can be administered once a day or divided into 2 to 4 times a day. In the case of transdermal administration, it is usually rubbed directly into the skin several times a day, for example, 1 to 5 times, an appropriate amount, for example, 0.1 to 1 ml per 1 cm ² , or the appropriate amount is soaked in gauze etc. Can be affixed and used.

  The whitening agent according to the present invention is particularly preferably applied in the form of an external preparation for skin, of cosmetics and pharmaceutical compositions, which is applied or sprayed directly to the skin or soaked in gauze or the like. It is used by the administration method that is applied to the skin and exhibits an excellent whitening effect.

  EXAMPLES The present invention will be specifically described with reference to examples, but the technical scope of the present invention is not limited to these examples. The blending amount is mass% unless otherwise specified.

[Preparation of Mannentake extract]
[Examples 1 to 10, Comparative Examples 1 to 7]
50 g of pulverized dried product of Kakuno Reishi fruit body was put into the extraction tank of the supercritical fluid extraction apparatus, and extracted with supercritical carbon dioxide at the temperature and pressure shown in Table 1. The extract obtained by being separated from carbon dioxide in the separation tank of the apparatus was dissolved in ethanol, recovered from the separation tank, dried under reduced pressure, and weighed. The extraction efficiency was determined from the amount of extract recovered for 50 g of the input raw material, and is shown in Table 1.

  From Table 1, by extracting with supercritical carbon dioxide in a pressure range of 45-50 MPa and a temperature range of 35-80 ° C., the extraction efficiency is significantly higher than that of a supercritical carbon dioxide pressure of 40 MPa or less. It is high and it turns out that extraction efficiency is critically good. Therefore, when extract the bamboo shoots with a fluid in a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa, particularly supercritical carbon dioxide in a temperature range of 30.9 to 150 ° C. and a pressure range of 42 to 100 MPa. It is clear that the extraction efficiency is greatly improved and the critical effect is obtained.

[Example 11] Whitening effect test [Inhibition of melanin production in B16 melanoma cells]
The melanin production inhibitory effect was evaluated by using B16 melanoma cells, which are mouse-derived melanocytes, and observing the production of melanin by acting on the cells with a solution obtained by diluting the specimen with a culture solution. B16 cells were cultured using 10% FBS E-MEM medium. The samples evaluated were those obtained by the methods of Examples and Comparative Examples. In addition, Kazuno Ganoderma ethanol extract was used for comparison. Details are described below.

[Experimental operation]
[Preparation of sample to be added to culture medium]
An ethanol solution having a concentration of 2 mg / ml was prepared for the Kazuno Ganoderma supercritical carbon dioxide extraction powder extracted under various conditions. For comparison, an ethanol-extracted powder of kakukaku reishi ground (200 mesh) was prepared as an ethanol solution having the same concentration (2 mg / ml). As a control, ethanol as a solvent was prepared. These were diluted as a stock solution in a medium or a medium containing 10% ethanol, and a dilution series was prepared so that each contained 10% ethanol. 20 μl of each diluted sample prepared was added to cells in culture. Since ethanol is toxic to cells, the final concentration in the culture solution was adjusted to 1% or less.

[Culture operation]
B16 cells previously cultured to about 70% confluence are peeled and collected, counted, and a cell suspension is prepared so that the cell density after seeding / adhesion is 5.0 × 10 ³ cells / cm ² , 180 μl each was seeded on a 96-well plate. After culturing for 3 to 5 hours, it was confirmed with a microscope that the cells were adhered, and 20 μl of sample solution or solvent alone (control) was added. 72 hours after addition of the sample solution or solvent, the accumulation state of black pigment in each cell was observed with an inverted microscope. By comparison with control cells, the amount of accumulated pigment was evaluated in three stages as shown below.

2: The amount of dye in the cell is clearly smaller than the control.
1: It is recognized that the amount of dye in the cell is smaller than that of the control.
0: The amount of intracellular dye is almost the same as the control.

  At the same time, the number of cells was visually compared with a microscope to confirm whether or not there was a great difference from the control.

[result]
The results of evaluating the ability to suppress melanin production for each sample in Table 1 are shown in Tables 2 and 3. The cell number was comparable to the control by visual observation with a microscope, or no significant difference was observed. Therefore, the cytotoxicity range was considered to be low over the range of concentrations tested.

  As shown in Tables 2 and 3, the Kano Ganoderma extract obtained in Examples 1 to 10 has an excellent inhibitory effect on melanin production on B16 melanoma cells, and is used as a whitening agent (component) and as a whitening agent. It turns out that it is useful. In addition, the deer antaceae extract obtained by Examples 1-10 has the same or better melanin production inhibitory effect in comparison with the deer antler retorii ethanol extract and the comparative samples obtained by Comparative Examples 1-7. You can see that

  From the above results, high extraction efficiency was observed under a pressure of 45 MPa or more, and the melanin production inhibitory activity of these extracts was high. Therefore, it is useful as a whitening agent (component) and useful as a whitening agent by performing an extraction treatment of Kakuno Reishi using carbon dioxide in a pressure range of 42 to 100 MPa and a temperature range of 30 to 150 ° C. It is clear that the extract can be obtained efficiently.

  The formulation example of the whitening agent based on this invention is shown below. In addition, what was obtained by the method of the said Example was used for the blended Kazuno Ganoderma extract.

[Formulation Example 1] Cream component blending amount (% by mass)
Stearic acid 6.0
Glycerin monostearate ester 2.0
POE (20 mol) sorbitan monostearate 2.0
1,3-butylene glycol 10.0
Kazuno Reishi Extract (Example 6) 1.0
Isopropyl myristate 12.0
Squalane 5.0
Liquid paraffin 3.0
Vitamin E 0.05
Sodium bisulfite Appropriate amount of preservative Appropriate amount of perfume Appropriate amount of ion-exchanged water Residue

[Formulation Example 2] Emulsion component content (mass%)
Stearyl alcohol 2.0
Squalane 5.0
Liquid lanolin 3.0
Trimethylol propane trioctanoate 2.0
Glyceryl trioctanoate 5.0
Glycerol monooleate 2.0
POE (20 mol) sorbitan monooleate 1.0
Vitamin C 0.1
Preservative Appropriate amount of Kakukaku Reishi extract (Example 7) 2.0
Paraaminobenzoic acid 0.1
Perfume Appropriate amount Sodium bisulfite Appropriate amount Glycerin 5.0
Carboxyvinyl polymer 0.2
Triethanolamine 1.0
Purified water residue

[Formulation Example 3] Gel component blending amount (% by mass)
Dipropylene glycol 10.0
PEG 1500 5.0
Carboxyvinyl polymer 1.0
Ethyl alcohol 2.0
POE (50 mol) oleyl alcohol ether 2.0
Potassium hydroxide 0.15
Kazuno Reishi Extract (Example 8) 0.5
Sodium 2-hydroxy-4-methoxybenzophenone sulfonate 0.05
Preservative Appropriate amount EDTA / 3Na Appropriate perfume Appropriate amount Ion-exchanged water Residue

[Formulation Example 4] Essence component Blending amount (% by mass)
Glycerin 5.0
Dipropylene glycol 10.0
Ethanol 10.0
Carboxyvinyl polymer 0.2
Sodium hyaluronate 0.5
Potassium hydroxide 0.1
POE (20 mol) sorbitan monooleate 0.5
POE (20 mol) octyldodecanol ether 1.0
Olive oil 0.15
Vitamin E acetate 0.1
Kazuno Reishi Extract (Example 9) 3.0
Sodium bisulfite Appropriate amount Preservative Appropriate amount EDTA / 3Na Appropriate perfume Appropriate amount Purified water Residue

[Formulation Example 5] Pack component amount (% by mass)
Polyvinyl acetate emulsion 15.0
Polyvinyl alcohol 10.0
Dipropylene glycol 5.0
Sorbitol 5.0
Tocopherol acetate 0.2
Olive oil 1.0
Squalane 3.0
POE (20 mol) sorbitan monooleate 1.5
Titanium oxide 5.0
Talc 10.0
Ethanol 7.0
Kazuno Reishi Extract (Example 10) 2.0
Preservative Appropriate amount of perfume Appropriate amount of purified water Residue

[Formulation Example 6] lotion component content (mass%)
1,3-butylene glycol 6.0
Glycerin 5.0
Sodium hyaluronate 0.3
POE (20 mol) sorbitan monostearate 1.5
Polyoxyethylene (20 mol) octyldodecanol ether 0.5
Ethanol 15.0
Kazuno Reishi Extract (Example 1) 0.1
Preservative Appropriate amount of perfume Appropriate amount of purified water Residue

[Formulation Example 7] Ointment component content (mass%)
POE (30 mol) hydrogenated castor oil 2.0
Glyceryl monostearate 10.0
Squalane 10.0
Vaseline 40.0
Cetanol 6.0
Kazuno Reishi Extract (Example 2) 5.0
1,3-butylene glycol 10.0
Ion-exchange water Residual preservative Appropriate amount Perfume Appropriate amount

[Formulation Example 8] Foundation component content (mass%)
Talc 3.0
Titanium oxide 5.0
Bengala 0.5
Iron oxide yellow 1.5
Iron oxide black 0.1
Bentonite 0.5
POE (20 mol) sorbitan monostearate 1.0
Triethanolamine 1.0
Glycerin 5.0
Propylene glycol 5.0
Stearic acid 2.0
Glycerol monostearate 10.0
Liquid paraffin 10.0
Dimethylpolysiloxane 5.0
Polyoxyalkylene-modified polysiloxane 3.0
Kazuno Reishi Extract (Example 3) 1.0
Purified water Residual preservatives

[Formulation Example 9] Tablet component content (mass%)
Kazuno Reishi Extract (Example 4) 5.0
Reduced maltose 59.0
Trehalose 14.0
Glycerin fatty acid ester 2.0
Dextrin 20.0

[Formulation Example 10] Candy component blending amount (mass%)
Sucrose 68.0
Minamata 30.0
Kazuno Reishi Extract (Example 5) 1.0
Fragrance 1.0

[Formulation Example 11] Granule component content (mass%)
Reduced maltose 35.0
Lactose 20.0
Crystalline cellulose 31.0
Kazuno Reishi Extract (Example 1) 12.0
Sugar Ester 2.0

[Formulation example 12] Tablet confectionery component blending amount (% by mass)
Kazuno Reishi Extract (Example 6) 2.0
Reduced maltose 20.0
Erythritol 10.0
Citric acid 10.0
Blueberry extract 5.0
Lactose 5.0
Sucrose fatty acid ester 3.0
Perfume appropriate amount dextrin

[Formulation Example 13] Beverage component content (mass%)
Kazuno Reishi Extract (Example 10) 0.1
Reduced maltose starch syrup 5.0
Erythritol 1.0
Vitamin C 1.0
Stevia 0.05
Fragrance 0.1
Water residue

Claims (6)

  A method for extracting nanentake extract having an inhibitory action on melanin production from nanentake, wherein the nanentake is extracted with a fluid in a temperature range of 30 to 150 ° C. and a pressure range of 42 to 100 MPa. A method for extracting a garlic mushroom extract having a production inhibitory action.   2. The method for extracting nanentake extract having an inhibitory action on melanin production according to claim 1, wherein the nanentake is deers.   3. The method for extracting mannentake extract having melanin production inhibitory action according to claim 1 or 2, wherein the fluid is carbon dioxide.   The whitening agent containing the mannentake extract extracted by the method as described in any one of Claims 1 thru | or 3.   The whitening agent according to claim 4, wherein the mannentake is kano ganshi.   The whitening agent according to claim 4 or 5, wherein the fluid is carbon dioxide.