Application of premna microphylla extract in preparation of cosmetics and cosmetic raw materials

Abstract

The invention relates to the technical field of effects of cosmetic raw materials, in particular to application of a premna microphylla extract in preparation of cosmetics and cosmetic raw materials. The invention provides an application of a premna microphylla extract in preparation of cosmetics with whitening and / or moisturizing effects, and the result of the embodiment shows that a premna microphylla pectin crude extract has good freckle-removing and whitening capabilities. The premna microphylla ethanol extract, the separation product of the ethanol extract and the premna microphylla pectin crude extract all show certain moisturizing capacity. Meanwhile, the moisturizing capacity of a petroleum ether phase of the premna microphylla ethanol extract is outstanding and exceeds the moisturizing level of sodium alginate, which is not involved in the existing research, and the premna microphylla ethanol extract has important significance for preventing oxidation-related diseases and developing moisturizing cosmetics. By combining the characteristics of abundant resources and low price of premna microphylla in China, the development of the premna microphylla meets the market demand of natural green cosmetic raw materials, and has sustainable development economic benefits.

Description

Technical Field

[0001] The invention relates to the technical field of the efficacy of cosmetic raw materials, and in particular to an application of a tofu chai extract in the preparation of cosmetics and a cosmetic raw material. Background Art

[0002] Premna Microphylla Turcz. is a plant of the genus Premna Turcz. of the Verbenaceae family. It is also known as Guanyin Chai, stinking yellow vitex, native astragalus (Zhejiang), Manshanxiang, hemostatic grass, etc. It grows mostly on hillsides or under forests, and is mainly distributed in tropical and subtropical regions. In my country, it is mainly produced in Hunan, Hubei, Sichuan and Guizhou provinces, and is an important plant for both medicinal and edible purposes. The roots, stems and leaves of Premna Turcz. contain a large amount of pectin, crude fiber, crude protein, and rich minerals and polyphenols, sterols, carotene, naringenin, vanillic acid and other medicinal ingredients. Therefore, Premna Turcz. has high nutritional and health-care effects and special medicinal uses. Premna Turcz. plants are widely distributed in my country, but they lack perfect cosmetic application development.

[0003] Existing studies have shown that the extract of tofu shiitake mushroom has anti-inflammatory, antioxidant and other effects. In the prior art, there is no application of tofu shiitake mushroom extract in the preparation of cosmetics with whitening and moisturizing effects. Summary of the invention

[0004] The present invention provides an application of a tofu taro extract in preparing cosmetics with whitening and / or moisturizing effects ， The prepared cosmetics have moisturizing and whitening effects.

[0005] In order to solve the above technical problems, the following technical solutions are proposed:

[0006] The present invention provides an application of a tofu bud extract in preparing cosmetics with whitening and / or moisturizing effects. The tofu bud extract comprises one or more of an alcohol extract of tofu bud, a crude pectin extract of tofu bud, an ethyl acetate phase extract of the alcohol extract of tofu bud, and a petroleum ether phase extract of the alcohol extract of tofu bud.

[0007] Preferably, the extractant used for the tofu taro extract includes ethanol and / or water with a volume concentration of 75%; when the extractant is ethanol with a volume concentration of 75%, tofu taro alcohol extract is obtained; when the extractant is water, tofu taro pectin crude extract is obtained;

[0008] The tofu bud chai alcohol extract is extracted by a system solvent extraction method to obtain the tofu bud chai alcohol extract ethyl acetate phase extract, the tofu bud chai alcohol extract petroleum ether phase extract and the tofu bud chai alcohol extract n-butanol phase extract.

[0009] Preferably, the extractants used in the system solvent extraction method include petroleum ether, ethyl acetate and n-butanol in sequence.

[0010] Preferably, the preparation method of the tofu chai alcohol extract comprises:

[0011] The tofu-tree crushed material is subjected to reflux extraction using 75% ethanol by volume to obtain a reflux extract; the reflux extract is filtered and concentrated to obtain tofu-tree alcohol extract; the reflux extraction is performed 2 to 4 times; the reflux extraction temperature is 75 to 85°C; and the time for each reflux extraction is 2 to 4 hours.

[0012] Preferably, during the reflux extraction, the solid-liquid ratio of the tofu-tree crushed material to 75% ethanol by volume is 1 g: (5-20) mL.

[0013] Preferably, the method for preparing the crude extract of shiitake mushroom pectin comprises: ultrasonically extracting shiitake mushroom leaves with water to obtain an extract; and sequentially concentrating, precipitating and decolorizing the extract to obtain the crude extract of shiitake mushroom pectin.

[0014] Preferably, the ultrasonic extraction time is 50 to 80 minutes; the ultrasonic extraction power is 20 to 50 kHz, and the extraction temperature is 80 to 95°C.

[0015] Preferably, the solvent used for the precipitation includes anhydrous ethanol.

[0016] The invention provides an application of an ethyl acetate phase of a tofu bud chai alcohol extract in preparing a product with moisturizing and / or antioxidant effects. The tofu bud chai alcohol extract is extracted with ethyl acetate to obtain the ethyl acetate phase of the tofu bud chai alcohol extract.

[0017] The present invention provides a tofu-wood cosmetic raw material, when the cosmetic raw material has the effect of whitening, it contains tofu-wood pectin crude extract;

[0018] When the cosmetic raw material has the function of moisturizing, it contains one or more of taro ethanol extract, taro pectin crude extract, ethyl acetate phase extract of taro ethanol extract and petroleum ether phase extract of taro ethanol extract;

[0019] When the cosmetic raw material has an antioxidant effect, it contains one or more of the following: tofu bud ethanol extract, tofu bud ethanol extract ethyl acetate phase extract and tofu bud ethanol extract petroleum ether phase extract.

[0020] Beneficial effects of the present invention: The present invention provides an application of a tofu bud extract in the preparation of cosmetics with whitening and / or moisturizing effects. The results of the embodiments show that the crude tofu bud pectin extract exhibits good freckle removal and whitening abilities. The tofu bud alcohol extract, the separated products of the alcohol extract, and the crude tofu bud pectin extract all exhibit certain moisturizing abilities. At the same time, the moisturizing ability of the petroleum ether phase extract of the tofu bud alcohol extract is relatively outstanding, exceeding the moisturizing level of sodium alginate. This is a field that has not been covered by existing research and is of great significance for preventing oxidation-related diseases and developing moisturizing cosmetics. Considering the fact that tofu bud is rich in resources and low in price in my country, its development not only meets the market demand for natural green cosmetic raw materials, but also has the economic benefits of sustainable development. In summary, the tofu bud extract has whitening and moisturizing effects and can be used to prepare cosmetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the DPPH free radical scavenging rate diagram of the alcohol extract of tofu chai;

[0022] Figure 2 This is the ABTS free radical scavenging rate diagram of tofu chai alcohol extract;

[0023] Figure 3 It is the moisture retention rate of crude extract of tofu chai pectin (i.e. water extract);

[0024] Figure 4 This is the tyrosinase clearance rate diagram of tofu chai ethanol extract (i.e., the tyrosinase inhibition rate diagram);

[0025] Figure 5 This is the ABTS free radical scavenging rate diagram of different polarity parts of tofu chai alcohol extract;

[0026] Figure 6 This is a graph showing the DPPH radical scavenging rate of different polar parts of the tofu bud alcohol extract. DETAILED DESCRIPTION

[0027] The present invention provides an application of a tofu bud extract in cosmetics with whitening and / or moisturizing effects. The tofu bud extract comprises one or more of an alcohol extract of tofu bud, a crude pectin extract of tofu bud, an ethyl acetate phase extract of the alcohol extract of tofu bud, and a petroleum ether phase extract of the alcohol extract of tofu bud.

[0028] As an optional implementation manner, the extraction part of the Premna microphylla Turcz. of the present invention is the Premna microphylla Turcz. leaves; the present invention has no special limitation on the source of the Premna microphylla Turcz. leaves, and conventional products can be used.

[0029] As an optional embodiment, the extractant used in the tofu taro extract of the present invention includes ethanol and / or water with a volume concentration of 75%; when the extractant is ethanol with a volume concentration of 75%, tofu taro alcohol extract is obtained; when the extractant is water, tofu taro pectin crude extract is obtained;

[0030] The tofu bud alcohol extract is extracted by a system solvent extraction method to obtain an ethyl acetate phase extract of the tofu bud alcohol extract, a petroleum ether phase extract of the tofu bud alcohol extract and a n-butanol phase extract of the tofu bud alcohol extract; the extractants used in the system solvent extraction method include petroleum ether, ethyl acetate and n-butanol in sequence.

[0031] As an optional embodiment, the method for preparing the alcohol extract of Fructus dasyphyllae leaves of the present invention comprises:

[0032] The tofu bud leaf crushed material is subjected to reflux extraction with 75% ethanol by volume to obtain a reflux extract; the reflux extract is concentrated to obtain the tofu bud alcohol extract. As an optional embodiment, the tofu bud leaf crushed material of the present invention is passed through a 60-mesh sieve, and the material under the sieve is taken; the tofu bud leaf crushed material is a dried tofu bud leaf crushed material.

[0033] As an optional embodiment, the solid-liquid ratio of the dried pulverized tofu leaves to 75% ethanol by volume during the reflux extraction is 1g: (5-20)mL, more preferably 1g: 10mL. In the present invention, the 75% ethanol helps to extract flavonoids, polyphenols and other components of tofu.

[0034] As an optional embodiment, the temperature for reflux extraction of the tofu bud leaf crushed material with 75% volume concentration ethanol is 75-85°C, preferably 80°C; the time for each reflux extraction is 2-4h, preferably 3.4h.

[0035] As an optional embodiment, when preparing the alcohol extract of the tofu bud leaf, a reflux extract is obtained after each reflux extraction, and the remaining filter residue of the reflux extraction is refluxed again to obtain a reflux extract. The number of reflux extractions in the present invention is 2 to 4 times, preferably 3 times. The volume of 75% ethanol added in each reflux extraction of the present invention is the same. In a specific embodiment of the present invention, the number of reflux extractions is 3 times.

[0036] After the reflux extraction, the reflux extracts are combined and concentrated to obtain the alcohol extract of the tofu cypress leaves. The concentration in the present invention is evaporation concentration, and the evaporation concentration is performed until the obtained product is dried to a constant weight state.

[0037] The alcohol extract of tofu buds prepared by the present invention is a crude extract, and the alcohol extract of tofu buds extracted by using a specific organic solvent of 75% ethanol combined with specific extraction temperature and time conditions can improve the antioxidant and moisturizing effects of the alcohol extract. The alcohol extract of tofu buds has a strong concentration dependence, and its antioxidant activity can be fully manifested at a higher concentration, which provides a good basis for its application in the development of antioxidant health food.

[0038] The present invention utilizes a systemic solvent extraction method to extract the tofu bud alcohol extract, and obtains an ethyl acetate phase extract of the tofu bud alcohol extract, a petroleum ether phase extract of the tofu bud alcohol extract, and a n-butanol phase extract of the tofu bud alcohol extract; the extractants used in the systemic solvent extraction method include petroleum ether, ethyl acetate, and n-butanol in sequence. As an optional embodiment, the tofu bud alcohol extract of the present invention is in the form of a dry powder; as an optional embodiment, the present invention first mixes the tofu bud alcohol extract with water and then extracts. The volume ratio of the tofu bud alcohol extract to water is 1g: (10-40)mL, preferably 1g: 20mL. The present invention records the mixture of the alcohol extract of tofu bud and water as a mixed liquid; as an optional embodiment, the present invention first uses petroleum ether to extract the mixed liquid with petroleum ether to obtain a petroleum ether layer extract and a water layer, and the volume ratio of the petroleum ether to the mixed liquid is 1:1 during the first petroleum ether extraction; the petroleum ether layer extract is separated and petroleum ether is added to the water layer again for a second petroleum ether extraction, and the petroleum ether layer extract and the water layer are obtained again; the present invention adds petroleum ether extractant multiple times until the petroleum ether layer extract is colorless, and the petroleum ether extraction is completed. The present invention mixes all the obtained petroleum ether layer extracts to obtain a petroleum ether phase extract; the volume of petroleum ether added in each extraction of the present invention is the same. The petroleum ether phase extract of the alcohol extract of tofu bud has the effects of moisturizing and whitening.

[0039] The present invention records the water layer obtained by the last extraction of petroleum ether as the first water layer, adds ethyl acetate to the first water layer for the first ethyl acetate extraction, and obtains an ethyl acetate layer extract and a water layer; the volume ratio of ethyl acetate to the mixed solution (i.e., the volume sum of the dry powder of the total alcohol extract of tofu taro and water) during the first ethyl acetate extraction of the present invention is 1:1, and the extraction method is the same as the petroleum ether extraction method. The present invention adds ethyl acetate extractant to the water layer for many times until the ethyl acetate layer extract is colorless, and the ethyl acetate extraction is finished. The present invention mixes all the obtained ethyl acetate layer extracts to obtain an ethyl acetate phase extract; the volume of ethyl acetate added in each extraction of the present invention is the same. The ethyl acetate phase extract of the alcohol extract of tofu taro leaves exhibits a better antioxidant capacity. In the DPPH free radical scavenging experiment, the ethyl acetate phase extract exhibits the strongest scavenging capacity, and at a high concentration, it is close to 100.00% as vitamin C (i.e., VC). In the ABTS free radical scavenging rate detection, the ethyl acetate phase extract also performs best, and at a high concentration, it is close to or reaches 100.00%, which is similar to the standard control VC.

[0040] The present invention records the water layer obtained in the last extraction with ethyl acetate as the second water layer, adds n-butanol to the second water layer for the first n-butanol extraction, and obtains the n-butanol layer extract and the water layer; in the first n-butanol extraction of the present invention, the volume ratio of n-butanol to the mixed solution (i.e. the volume sum of the dry powder of the total alcohol extract of tofu chai and water) is 1:1, and the extraction method is the same as the petroleum ether extraction method. The present invention adds n-butanol extractant to the water layer for multiple times until the n-butanol layer extract is colorless, and the n-butanol extraction is finished.

[0041] The research on the cosmetic efficacy of different polar parts of the alcohol extract of tofu buds is still insufficient at home and abroad, and there is a large space for exploration. There is no research on the efficacy of different polar parts separated by the solvent extraction method of tofu buds. The ethyl acetate phase extract of the alcohol extract of tofu buds obtained by the present invention has good antioxidant capacity and moisturizing performance. The petroleum ether phase extract of the alcohol extract of tofu buds has the effects of moisturizing and whitening. The present invention adopts different separation methods and uses solvents of different polarities for extraction. The product after the alcohol extract of tofu buds shows better efficacy. The antioxidant activity of the ethyl acetate phase extract of the alcohol extract of tofu buds is close to that of the natural antioxidant vitamin C, which can effectively remove free radicals and alleviate cell damage and aging caused by oxidation.

[0042] As an optional embodiment, the preparation method of the crude extract of tofu taro leaf pectin of the present invention comprises: ultrasonically extracting tofu taro leaf with water to obtain an extract; sequentially concentrating, precipitating and decolorizing the extract to obtain a crude extract of tofu taro pectin. The crude extract of tofu taro pectin is an aqueous extract of tofu taro leaf; the tofu taro leaf is dried before ultrasonic extraction, and the present invention has no special limitation on the drying method, and a conventional method can be used. The water added during the ultrasonic extraction of the present invention can cover the tofu taro leaf. The ultrasonic extraction time of the present invention is 50 to 80 minutes, preferably 70 minutes; the power of the ultrasonic extraction is 20 to 50 kHz, preferably 40 kHz; the temperature of the ultrasonic extraction is 80 to 95°C, preferably 90°C. After the ultrasonic is finished, the ultrasonic extract is filtered to obtain an ultrasonic extract; the filtration is preferably gauze filtration; the present invention centrifuges the ultrasonic extract to obtain a supernatant; the supernatant is sequentially concentrated, precipitated, decolorized and dried to obtain crude tofu taro leaf pectin. The concentration method of the present invention is rotary evaporation concentration to obtain a concentrate; the concentration is completed to 2g / mL (2g of crude drug per milliliter). During the precipitation of the present invention, anhydrous ethanol is added to the concentrate and allowed to stand to obtain a precipitate; during the precipitation, the volume ratio of the concentrate to anhydrous ethanol is 1:2 to 1:4, preferably 1:3; the standing temperature is 4°C, and the standing time is overnight. The present invention uses 70% ethanol, 95% ethanol and anhydrous ethanol to wash the precipitate in turn to achieve decolorization. The number of washings with 70% ethanol, 95% ethanol and anhydrous ethanol in the present invention is preferably 2 times respectively. The crude extract of tofu bud pectin of the present invention has a whitening effect.

[0043] The present invention provides an application of an ethyl acetate phase extract of an alcohol extract of Tofu Chaihu in preparing a product with moisturizing and / or antioxidant effects, wherein the alcohol extract of Tofu Chaihu is extracted with ethyl acetate to obtain an ethyl acetate phase extract of an alcohol extract of Tofu Chaihu. The technical scheme of extracting the alcohol extract of Tofu Chaihu with ethyl acetate has been discussed above and will not be repeated here. The ethyl acetate phase extract of the alcohol extract of Tofu Chaihu leaves obtained by the present invention has good antioxidant capacity and moisturizing performance.

[0044] The present invention provides a tofu-wood cosmetic raw material, when the cosmetic raw material has the effect of whitening, it contains tofu-wood pectin crude extract;

[0045] When the cosmetic raw material has the function of moisturizing, it contains one or more of taro ethanol extract, taro pectin crude extract, ethyl acetate phase extract of taro ethanol extract and petroleum ether phase extract of taro ethanol extract;

[0046] When the cosmetic raw material has an antioxidant effect, it contains one or more of the following: tofu bud ethanol extract, tofu bud ethanol extract ethyl acetate phase extract and tofu bud ethanol extract petroleum ether phase extract.

[0047] As an optional embodiment, the present invention has no special restrictions on the preparation method and auxiliary materials of the cosmetics, and conventional methods and products can be used.

[0048] In order to further illustrate the present invention, the application of the tofu taro extract provided by the present invention and the cosmetics with whitening and / or moisturizing effects and the preparation method thereof are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0049] Example 1 Material extraction

[0050] 1 Extraction of crude tofu shiitake extract and separation of different polarity fractions

[0051] 1.1 Extraction of total alcohol extract of Tofu Chai

[0052] Take dried tofu bud leaf and crush it, then pass it through a 60-mesh sieve to obtain tofu bud leaf powder. Accurately weigh 100.00g of tofu bud leaf powder and heat it in a round-bottom flask for reflux extraction. The powder is dissolved in 1000mL of ethanol. The solid-liquid ratio of tofu bud leaf powder and 75% ethanol by volume is 1:10 (g / mL). Reflux conditions: 75% ethanol is heated to 80°C and refluxed for 3.4h, and the filtrate is collected by filtration. The obtained filter residue is added with 1000mL of 75% ethanol by volume for reflux extraction for the second time, and the filter residue obtained by the second reflux extraction is added with 1000mL of 75% ethanol by volume for reflux extraction for the third time. The temperature and time of the three reflux extractions are the same, and the three reflux filtrates are combined. The obtained filtrate is used to recover the solvent by rotary evaporation until dry, and the dry powder of the total alcohol extract of tofu bud leaf is obtained.

[0053] 1.2 Separation of different polarity fractions of total alcohol extract of tofu chai

[0054] 2.0 g of dried powder of total alcohol extract of tofu taro is taken, dissolved in 20 times the volume of water to obtain a mixed solution, and placed in an extraction separatory funnel. Petroleum ether is added in the extraction separatory funnel according to a volume ratio of the mixed solution to petroleum ether of 1:1 to obtain a petroleum ether layer and a water layer; the petroleum ether layer is separated, and petroleum ether is added to the water layer again for extraction to obtain a petroleum ether layer and a water layer again until the petroleum ether layer extract is colorless, and the petroleum ether extraction is terminated, and all the obtained petroleum ether layers are mixed to obtain a petroleum ether phase extract; the volume of petroleum ether added for each extraction is the same.

[0055] The water layer obtained in the last extraction with petroleum ether is recorded as the first water layer, and ethyl acetate is added to the first water layer for extraction. The volume ratio of the added ethyl acetate and the mixed solution (i.e., the volume sum of the total alcohol extract of tofu chai and water) is 1:1. The extraction method is the same as the petroleum ether extraction method, until the ethyl acetate layer extract is colorless, and the ethyl acetate extraction is terminated; all the ethyl acetate layers obtained are mixed to obtain an ethyl acetate phase extract, and the volume of ethyl acetate added for each extraction is the same.

[0056] The water layer obtained in the last extraction with ethyl acetate is recorded as the second water layer, and n-butanol is added to the second water layer for extraction. The volume ratio of the added n-butanol and the mixed solution (i.e., the volume of the total alcohol extract of tofu chai and water) is 1:1. The extraction method is the same as the petroleum ether extraction method, until the n-butanol layer extract is colorless, and the n-butanol extraction is completed. All the n-butanol layers obtained are mixed to obtain the n-butanol phase extract, and the volume of n-butanol added each time is the same. The water layer obtained in the last extraction with n-butanol is recorded as the third water layer, i.e., the aqueous phase extract.

[0057] 1.3 Extraction and purification of crude tofu pectin

[0058] Weigh 100g of dried tofu leaves powder and transfer it into a large beaker. Add an appropriate amount of distilled water to the large beaker until the dry powder is covered. After uniform stirring, place the large beaker in an ultrasonic cleaning machine with an ultrasonic power intensity of 40kHz for ultrasonic extraction, the ultrasonic time is 70min, and the ultrasonic temperature is 90°C. After the ultrasonic treatment, filter the extract with gauze to remove the residue, put it into a centrifuge and centrifuge it at 5000r / min for 15min, collect the supernatant and concentrate it with a rotary evaporator, add 3 times the volume of anhydrous ethanol after concentration, let it stand overnight at 4°C (room temperature), take the precipitate, wash it twice with 70% ethanol by volume, 95% ethanol by volume and anhydrous ethanol to decolorize it, place the obtained wet pectin in a 60°C drying oven to dry to constant weight, and obtain crude pectin after drying.

[0059] Example 2 Efficacy Determination

[0060] 1. Determination of cosmetic efficacy of tofu chai ethanol extract and tofu chai pectin crude extract

[0061] 1.1 Antioxidant efficacy determination

[0062] 1.1.1 DPPH free radical scavenging ability

[0063] Accurately weigh a certain mass of DPPH powder, use 75% volume concentration ethanol as solvent to prepare 0.04 mg / mL DPPH solution, and store it at 2-4°C.

[0064] The total alcohol extract of Fructus Tofu (i.e., ethanol extract) was prepared into sample solutions with concentrations of 0.08, 0.10, 0.20, 0.40, 0.60, 0.80, and 1.00 mg / mL, respectively, and the VC reference substance was prepared into sample solutions with different mass concentrations of 0.01, 0.02, 0.04, 0.08, 0.10, and 0.2 mg / mL.

[0065] 1 mL of sample solution was taken respectively, and 3 mL of 0.04 mg / mL DPPH solution was added, mixed, and reacted in the dark for 30 min. The absorbance was measured at a wavelength of 517 nm and recorded as A1. Then 1 mL of distilled water was used to replace the sample solution, and 3.0 mL of DPPH solution was added. The reaction was carried out at room temperature for 30 min under the same conditions. The absorbance was measured at a wavelength of 517 nm and recorded as A0. The DPPH radical scavenging rate was calculated according to the following formula.

[0066]

[0067] The results are shown in Table 1 and Figure 1 :

[0068] Table 1 DPPH free radical scavenging rate of alcohol extract

[0069] Concentration (mg / mL) 0.01 0.02 0.04 0.08 0.10 0.20 0.40 0.60 0.80 1.00 Ethanol extract removal rate (%) - - - 20.50 43.60 73.51 84.64 86.56 89.06 89.83 VC clearance rate (%) 37.47 47.63 66.47 89.22 90.73 100.00 - - - -

[0070] Note: “-” means the clearance rate at the corresponding concentration was not measured.

[0071] 1.1.2 ABTS free radical scavenging ability

[0072] Mix equal volumes of 2.6 mmol / L potassium persulfate solution and 7.4 mmol / L ABTS solution, let stand at room temperature in the dark for 12 to 16 hours to prepare an ABTS cation stock solution, and dilute it with a phosphate buffer solution with a pH of 6.8 to an absorbance of 0.70±0.01 at 734 nm.

[0073] The alcohol extract of Dauci tataricus was prepared into sample solutions with different mass concentrations of 0.10, 0.20, 0.40, 0.60, 0.80, and 1.00 mg / mL, and the VC reference substance was prepared into sample solutions with different mass concentrations of 0.01, 0.02, 0.04, 0.08, 0.10, and 0.20 mg / mL.

[0074] 1 mL of sample solution and 6 mL of ABTS cation solution were respectively taken into test tubes, mixed thoroughly and reacted in the dark for 6 minutes, and the absorbance was measured at a wavelength of 734 nm, which was recorded as B1; distilled water was used instead of sample solution, and the absorbance was measured at a wavelength of 734 nm, which was recorded as B0; distilled water was used instead of ABTS cation solution, which was recorded as B2; ABTS clearance was calculated according to the following formula, and ascorbic acid (VC) was used as a control.

[0075]

[0076] The results are shown in Table 2 and Figure 2 .

[0077] Table 2 ABTS free radical scavenging rate of alcohol extract

[0078]

[0079]

[0080] Note: “-” means the clearance rate at the corresponding concentration was not measured.

[0081] It can be seen that the alcohol extract of tofu bud and VC (ascorbic acid) both showed certain ABTS free radical scavenging ability at different concentrations. With the increase of concentration, the free radical scavenging rate of the alcohol extract gradually increased, approaching the maximum scavenging rate of VC. However, the scavenging ability of the alcohol extract at lower concentrations was relatively weak, significantly lower than the scavenging rate of VC at the same concentration. This shows that the antioxidant effect of the alcohol extract is limited at low concentrations, but its scavenging ability at high concentrations can be significantly enhanced and approach the level of the standard antioxidant VC. This result shows that the alcohol extract of tofu bud has a strong concentration dependence, and its antioxidant activity needs to be fully reflected at higher concentrations, which provides a good basis for its application in the development of antioxidant health foods.

[0082] According to Tables 1-2 and Figures 1-2 It can be seen that the scavenging ability of the total alcohol extract of tofu taro (i.e., alcohol extract) for DPPH free radicals is generally better than its scavenging ability for ABTS free radicals, and it shows a higher scavenging efficiency. Compared with VC, the scavenging ability of alcohol extract at low concentrations of DPPH is also slightly inferior, but the scavenging rate gap in the medium and high concentration range is gradually narrowed, showing good antioxidant potential. This shows that tofu taro alcohol extract not only has a high efficiency in scavenging DPPH free radicals, but also has a certain concentration adaptability, which can meet the requirements of a high-efficiency antioxidant.

[0083] 1.2 Determination of moisturizing efficacy

[0084] 1.2.1 Determination of moisture retention rate of tofu chai alcohol extract

[0085] Weigh the tofu taro alcohol extract sample and glycerol and sodium alginate dried to constant weight, accurately weigh them, put them into a weighing bottle, accurately add purified water corresponding to the sample mass, rotate the weighing bottle to mix evenly, put it into a desiccator filled with color-changing powder silica gel, and weigh them after leaving them for 2, 4, 6, 8, 10, 24, and 36 hours. Calculate the moisture retention rate according to formula (3). The results are shown in Table 3.

[0086]

[0087] Where: H0 and Hn are the masses before and after placement for the corresponding time, mg.

[0088] Sample preparation: The alcohol extract of Fructus Tofu was mixed with water to obtain alcohol extract solutions with mass concentrations of 1%, 3%, 6%, and 8%;

[0089] Mixing glycerol and water to obtain a glycerol solution with a mass concentration of 3%;

[0090] Sodium alginate and water are mixed to obtain a sodium alginate solution with a mass concentration of 1%.

[0091] Table 3 Alcohol extract moisture retention rate

[0092]

[0093]

[0094] It can be seen that the moisturizing properties of different concentrations of tofu chai alcohol extract at different time points show significant differences. Overall, the moisturizing effect of the alcohol extract gradually weakens with time, but it has good moisturizing ability in a short period of time (2h to 10h), among which the 60mg / mL alcohol extract performs most prominently. However, as time extends to 24h and 36h, the moisturizing effects of each group decrease significantly, indicating that high-concentration alcohol extracts have more advantages in long-term moisturizing. Compared with the control group of 3% glycerol and 1% sodium alginate, the overall moisturizing performance of the alcohol extract is slightly insufficient. In contrast, the alcohol extract has certain competitiveness in short-term moisturizing, especially the high-concentration group, but it still needs to be optimized in long-term moisturizing. This shows that tofu chai alcohol extract is more suitable for use in short-term rapid hydration products in cosmetics, rather than in formulas with long-term moisturizing requirements. Subsequent studies can improve its moisturizing durability by modifying the active ingredients or compounding with other moisturizers, thereby broadening its application range.

[0095] 1.2.2 Determination of moisture retention rate of crude tofu pectin extract

[0096] The determination method is the same as 1.2.1, the only difference is that the tofu taro alcohol extract sample is replaced by the tofu taro pectin crude extract sample. The tofu taro pectin crude extract was mixed with water to obtain pectin crude extract solutions with mass concentrations of 1%, 3%, 6%, and 8% for determination. The results are shown in Tables 4 and Figure 3 .

[0097] Table 4 Moisture retention rate of tofu wood pectin crude extract (%)

[0098] time 2h 4h 6h 8h 10h 24h 36h 1% crude pectin moisture retention rate (%) 96.17 89.55 84.46 78.72 74.45 38.45 19.49 3% coarse pectin moisture retention rate (%) 91.82 86.15 81.05 74.83 70.24 37.93 17.29 6% coarse pectin moisture retention rate (%) 96.61 91.20 86.34 81.86 77.87 41.17 38.81 8% coarse pectin moisture retention rate (%) 93.46 82.73 80.00 73.90 70.02 40.98 37.12

[0099] It can be seen that the crude extract of tofu shiitake pectin has a significant moisturizing effect at a concentration of 6%. The moisturizing effect of 6% crude extract of tofu shiitake pectin is significantly better than that of other experimental groups.

[0100] 1.3 Whitening efficacy determination - inhibition of tyrosinase activity

[0101] 1.3.1 Arbutin was used as a control to analyze the inhibitory effect of the alcohol extract of Tofu Chai on tyrosinase activity. The reaction system is shown in Table 5.

[0102] Table 5 Reaction solution composition and volume

[0103]

[0104]

[0105] Prepare pH 6.8 phosphate buffer;

[0106] Prepare the sample solutions of the alcohol extract of Dauci ciliata with the mass concentrations of 0.01, 0.05, 0.2, 0.5 and 0.8 mg / mL respectively;

[0107] Arbutin positive control solutions were prepared with mass concentrations of 0.01, 0.05, 0.2, 0.5 and 0.8 mg / mL;

[0108] Prepare a tyrosinase solution with a mass concentration of 200 μg / mL;

[0109] Use a pipette to add different concentrations of tofu tsai alcohol extract sample solution, phosphate buffer, different concentrations of arbutin positive control solution and tyrosinase solution to the test tube in turn according to the volume of the reaction solution composition and volume in Table 5. Place in a 37°C water bath and mix for 10 minutes. Then add the substrate 1mg / mL levodopa solution. After fully reacting for 20 minutes, quickly transfer to a cuvette and measure the absorbance value at a wavelength of 475nm. Each concentration experiment is performed in parallel 3 times. According to the following formula, the inhibition rate of tofu tsai alcohol extract on tyrosinase is calculated:

[0110]

[0111] Where A1 is the absorbance with enzyme but no sample, A2 is the absorbance without enzyme and no sample, B1 is the absorbance with enzyme and sample, B2 is the absorbance with sample without enzyme, C1 is the absorbance of positive control with enzyme, and C2 is the absorbance of positive control without enzyme. When calculating the inhibition rate of tyrosinase of positive control, replace B1 with C1 and replace B2 with C2. The results are shown in Table 6 and Figure 4 .

[0112] Table 6 Inhibition rate of tyrosinase by different concentrations of alcohol extract and arbutin

[0113] Concentration (mg / mL) 0.01 0.05 0.2 0.5 0.8 Tyrosinase clearance rate of alcohol extract (%) 13.25 16.09 18.30 18.61 19.56 Arbutin's clearance rate of tyrosinase (%) 66.58 87.39 89.38 91.27 97.38

[0114] It can be seen that the inhibitory effect of tofu taro alcohol extract on tyrosinase is relatively weak, and its clearance rate increases slightly with the increase of concentration, but the increase is limited. The alcohol extract can only achieve limited enzyme inhibition effect at higher concentrations, which is quite different from the standard control group arbutin. Although the tyrosinase inhibition effect of tofu taro alcohol extract is far inferior to arbutin, its gradually increasing clearance rate shows that the extract still has a certain inhibitory ability and may have potential in compound applications. Compared with arbutin, tofu taro alcohol extract has a wide source, low cost, and milder inhibitory effect, which may be suitable for whitening products with lower inhibition intensity requirements. Combined with its multiple functions such as antioxidant and moisturizing, the application value of the alcohol extract may be more suitable for positioning as a functional synergist rather than a single high-efficiency enzyme inhibitor.

[0115] 1.3.2 Inhibitory effect of crude extract of tofu taro pectin on tyrosinase activity. The determination method is the same as 1.3.1, the only difference is that the total alcohol extract of tofu taro (i.e., ethanol extract) is replaced by crude extract of tofu taro pectin. The results are shown in Table 7. Arbutin positive control solutions were prepared with mass concentrations of 0.01, 0.05, 0.2, 0.5 and 0.8 mg / mL respectively; the results of arbutin clearance rate on tyrosinase are shown in Table 6.

[0116] Table 7 Inhibition rate of tyrosinase by different concentrations of pectin crude extract

[0117]

[0118] The crude extract samples of tofu shiitake pectin showed excellent whitening effect.

[0119] 2. Determination of the efficacy of different polar extraction parts of alcohol extract

[0120] 2.1 ABTS clearance rate

[0121] The method for determining the ABTS clearance rate of the alcohol extract of tofu chai was the same as that in 1.1.2 of Example 2. The results are shown in Tables 8 and Figure 5 .

[0122] Table 8 ABTS free radical scavenging rate at different polarity sites of alcohol extract

[0123]

[0124] Note: “-” means the clearance rate at the corresponding concentration was not measured.

[0125] It can be seen that the scavenging ability of different polar parts of the alcohol extract on ABTS free radicals is significantly different, among which the ethyl acetate phase performs best, which is close to the standard control VC. The n-butanol phase is second, and the scavenging ability of the petroleum ether phase, dichloromethane phase and water phase is weak, especially the water phase has the weakest scavenging ability, indicating that its antioxidant activity is limited. This difference may be due to the different types and concentrations of antioxidant active components in each polar part, especially the ethyl acetate phase may be rich in phenolic or flavonoid compounds with strong antioxidant activity, showing extremely high free radical scavenging efficiency.

[0126] 2.2DPPH free radical scavenging rate

[0127] The DPPH free radical scavenging rate determination method is the same as 1.1.1 of Example 2. The results are shown in Tables 9 and Figure 6 .

[0128] Table 9 DPPH radical scavenging rate of different polarity parts of alcohol extract

[0129]

[0130] Note: “-” means the clearance rate at the corresponding concentration was not measured.

[0131] It can be seen that the antioxidant properties of different polar parts are different. In the DPPH free radical scavenging experiment, the ethyl acetate phase still showed the strongest scavenging ability, and n-butanol showed good antioxidant activity. In contrast, the scavenging abilities of the petroleum ether phase, dichloromethane phase, and water phase were still weak, indicating that the ethyl acetate phase performed consistently in the two antioxidant systems and may contain a variety of active substances that are sensitive to different free radicals, while the antioxidant components in other parts are relatively scarce.

[0132] 2.3 Moisture retention rate

[0133] The method for determining the moisture retention rate is the same as 1.2.1 of Example 2.

[0134] The results are shown in Table 10, which shows that the moisturizing properties of the petroleum ether phase and the ethyl acetate phase are relatively balanced, and the petroleum ether phase shows certain advantages especially in medium-term moisturizing.

[0135] Table 10 Moisture retention rate of different polarity parts of alcohol extract

[0136] Time (h) 2h 4h 6h 8h 10h 24h 36h Glycerin moisturizing rate (%) 93.40 86.70 81.40 75.60 72.80 45.20 24.10 Ethyl acetate phase moisture retention rate (%) 89.60 79.80 72.50 65.50 60.80 28.20 6.10 Petroleum ether phase moisture retention rate (%) 90.80 84.80 79.30 73.40 69.10 37.20 14.40 Sodium alginate moisture retention rate (%) 90.50 81.80 73.20 69.00 60.80 21.20 13.10

[0137] In summary, combined with the analysis of free radical scavenging ability and moisturizing performance, the ethyl acetate phase has outstanding antioxidant activity, especially in the ABTS and DPPH free radical scavenging experiments at high concentrations, its effect is close to or even comparable to the standard antioxidant VC, showing significant application potential. However, its moisturizing ability is good in a short period of time, but the long-term effect is obviously insufficient, which limits its direct application in long-lasting moisturizing products. In contrast, although the petroleum ether phase is inferior to the ethyl acetate phase in antioxidant performance, its moisturizing performance is relatively balanced, especially showing certain advantages in mid-term moisturizing. The dichloromethane phase performed weakly in both functional tests and may only have certain value in auxiliary antioxidant or moisturizing. Overall, the functional performance of each polar part of the alcohol extract is complementary, which provides ideas for subsequent research and development, that is, to optimize product performance through the synergistic compounding of multiple parts, so as to achieve a dual improvement in antioxidant and moisturizing effects.

[0138] In summary, the alcohol extract of tofu chai has a certain antioxidant capacity, but the ethyl acetate phase after its separation shows a better antioxidant capacity. In the DPPH free radical scavenging experiment, the ethyl acetate phase shows the strongest scavenging ability, which is close to 100.00% at high concentrations, just like VC. In the ABTS free radical scavenging rate detection, the ethyl acetate phase also performs best, approaching or reaching 100.00% at high concentrations, which is similar to the standard control VC. The present invention adopts different separation methods and uses solvents of different polarities for extraction. The product after the alcohol extract of tofu chai is separated shows a better effect, and its antioxidant activity is close to the natural antioxidant vitamin C, which can effectively scavenge free radicals and alleviate cell damage and aging caused by oxidation.

[0139] The crude extract of tofu shiitake pectin of the present invention has a good inhibitory effect on tyrosinase, which is a key protein for the production of spots. It can be seen that the crude extract of tofu shiitake pectin exhibits good spot-removing and whitening capabilities.

[0140] The alcohol extract of tofu taro, its separation products and crude pectin all showed a certain moisturizing ability. At the same time, the moisturizing ability of the petroleum ether phase was more prominent, exceeding the moisturizing level of sodium alginate, which is an area that has not been covered by existing research and is of great significance for preventing oxidation-related diseases and developing moisturizing cosmetics. Considering the characteristics of tofu taro in my country, which is rich in resources and low in price, its development not only meets the market demand for natural green cosmetic raw materials, but also has sustainable economic benefits.

[0141] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A use of a tofu shiitake mushroom extract in preparing cosmetics with whitening and / or moisturizing effects, characterized in that: The tofu bud extract comprises one or more than two of tofu bud alcohol extract, tofu bud pectin crude extract, tofu bud alcohol extract ethyl acetate phase extract and tofu bud alcohol extract petroleum ether phase extract.

2. The use according to claim 1, characterized in that: The extractant used in the tofu taro extract includes ethanol and / or water with a volume concentration of 75%; when the extractant is ethanol with a volume concentration of 75%, tofu taro alcohol extract is obtained; when the extractant is water, tofu taro pectin crude extract is obtained; The tofu bud chai alcohol extract is extracted by a system solvent extraction method to obtain the tofu bud chai alcohol extract ethyl acetate phase extract, the tofu bud chai alcohol extract petroleum ether phase extract and the tofu bud chai alcohol extract n-butanol phase extract.

3. The application according to claim 2, characterized in that: The extractants used in the system solvent extraction method include petroleum ether, ethyl acetate and n-butanol in sequence.

4. The use according to claim 1 or 2, characterized in that: The preparation method of the tofu chai alcohol extract comprises: The tofu-tree crushed material is subjected to reflux extraction using 75% ethanol by volume to obtain a reflux extract; the reflux extract is filtered and concentrated to obtain tofu-tree alcohol extract; the reflux extraction is performed 2 to 4 times; the reflux extraction temperature is 75 to 85°C; and the time for each reflux extraction is 2 to 4 hours.

5. The use according to claim 4, characterized in that: During the reflux extraction, the solid-liquid ratio of the tofu shiitake powder to 75% ethanol by volume is 1 g: (5-20) mL 。 6. The use according to claim 2, characterized in that: The preparation method of the crude extract of tofu taro pectin comprises: using water to ultrasonically extract tofu taro leaves to obtain an extract; and sequentially concentrating, precipitating and decolorizing the extract to obtain the crude extract of tofu taro pectin.

7. The use according to claim 6, characterized in that: The ultrasonic extraction time is 50 to 80 minutes; the ultrasonic extraction power is 20 to 50 kHz, and the extraction temperature is 80 to 95°C.

8. The use according to claim 6, characterized in that: The solvent used for the precipitation includes anhydrous ethanol.

9. The use of the ethyl acetate phase of the alcohol extract of tofu chai in the preparation of a product with moisturizing and / or antioxidant effects, characterized in that: The tofu bud chai alcohol extract is extracted with ethyl acetate to obtain the tofu bud chai alcohol extract ethyl acetate phase.

10. A tofu-wood cosmetic raw material, characterized in that: When the cosmetic raw material has the effect of whitening, it contains crude extract of tofu taro pectin; When the cosmetic raw material has the function of moisturizing, it contains one or more of taro ethanol extract, taro pectin crude extract, ethyl acetate phase extract of taro ethanol extract and petroleum ether phase extract of taro ethanol extract; When the cosmetic raw material has an antioxidant effect, it contains one or more of the following: tofu bud ethanol extract, tofu bud ethanol extract ethyl acetate phase extract and tofu bud ethanol extract petroleum ether phase extract.

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