Anti-aging and whitening pyracantha fortuneana fruit extract prepared by adopting supramolecular solvent and preparation method of pyracantha fortuneana fruit extract

Abstract

The invention provides an anti-aging and whitening pyracantha fortuneana fruit extract prepared by adopting a supramolecular solvent and a preparation method of the pyracantha fortuneana fruit extract, and relates to the technical field of biological extraction. The preparation method comprises the following steps: by taking betaine as a hydrogen bond acceptor and 1, 3-propylene glycol as a hydrogen bond donor, mixing and self-assembling the betaine and the 1, 3-propylene glycol to form a supramolecular solvent; and the pyracantha fortuneana fruits are extracted by adopting the supramolecular solvent. The pyracantha fortuneana fruit active ingredients are extracted by adopting the supramolecular solvent which is harmless to the skin, so that the irritation of the solvent to the skin can be avoided, the supramolecular solvent can be specifically combined with the active ingredients in the pyracantha fortuneana fruit, and the purity and activity of the active ingredients are improved. The pyracantha fortuneana fruit extract extracted by adopting the supramolecular solvent has better oxidation reduction resistance; generation of non-enzymatic glycosylation intermediate products is inhibited, and saccharification and aging resistance are facilitated; the red blood cell oxidative damage is improved, and the effects of resisting oxidation and relieving are achieved; the whitening mask has an inhibiting effect on elastinase and tyrosinase, and can resist aging, whiten skin, relieve facial pigmentation and brighten the skin.

Description

Technical Field

[0001] The invention relates to the technical field of biological extraction, and in particular to an anti-aging and whitening Pyracantha fortuneana fruit extract prepared by adopting a supramolecular solvent and a preparation method thereof. Background Art

[0002] With the improvement of people's living standards and the increasing attention paid to their own health and appearance, anti-aging and skin whitening have become hot topics in the field of beauty and skin care. Skin aging is mainly caused by natural aging and photoaging. During the natural aging process, the metabolism of skin cells slows down, and the synthesis of collagen and elastic fibers decreases, resulting in thinning, sagging, and increased wrinkles on the skin. Photoaging is caused by long-term exposure to environmental factors such as ultraviolet rays, which triggers oxidative stress reactions and produces a large number of free radicals, further damaging the DNA, proteins and lipids of skin cells, and accelerating the aging process. At the same time, the excessive production and deposition of melanin is the main cause of dull skin and the formation of spots, which affects the whitening effect of the skin. Therefore, the development of safe and effective anti-aging and whitening skin care ingredients is of great practical significance.

[0003] In the current beauty and skin care market, common anti-aging ingredients such as retinoids, vitamin C, peptides, etc., can improve skin aging symptoms to a certain extent, but retinoids may cause adverse reactions such as skin irritation and dryness; vitamin C has poor stability and is easily oxidized and ineffective. As for whitening ingredients such as hydroquinone and kojic acid, hydroquinone has been restricted in some countries due to its potential cytotoxicity, and kojic acid also has problems such as poor stability and the possibility of allergic reactions.

[0004] Pyracantha fruit is rich in a variety of bioactive ingredients, such as flavonoids, polysaccharides, polyphenols, etc. Studies have shown that flavonoids have antioxidant effects, can remove free radicals in the body, and reduce the damage of oxidative stress to skin cells, thereby playing an anti-aging role; polyphenols can inhibit the activity of tyrosinase, reduce the synthesis of melanin, and have whitening potential. However, the current application of Pyracantha fruit extract in the field of beauty and skin care is not widespread, mainly because its extraction process is not perfect. Traditional extraction methods are difficult to fully extract the active ingredients in Pyracantha fruit, and there are many impurities in the extract, which affects the purity and stability of its active ingredients, limiting the application of Pyracantha fruit extract in high-end beauty and skin care products.

[0005] As a new type of green solvent, supramolecular solvent is an amphiphilic compound. It is a nanostructured micelle aggregate formed by an orderly self-assembly process between molecules. It has a unique microstructure and excellent performance. It can specifically bind to the target substance through weak intermolecular interactions (such as hydrogen bonds, van der Waals forces, etc.), thereby improving the extraction efficiency and selectivity of the target component. In addition, supramolecular solvents have good biocompatibility and environmental friendliness, overcoming many disadvantages of traditional organic solvents. In the field of natural product extraction, supramolecular solvents have gradually shown their application potential, but they have not been fully studied and applied in the preparation of Pyracantha fruit extract. Therefore, developing a method for efficiently preparing high-purity and high-activity anti-aging and whitening Pyracantha fruit extract using supramolecular solvents is of great significance to meet the market demand for safe and effective beauty and skin care ingredients and promote the widespread application of Pyracantha fruit in the field of beauty and skin care. Summary of the invention

[0006] The purpose of the present invention is to provide an anti-aging and whitening Pyracantha fortuneana fruit extract prepared by using a supramolecular solvent and a preparation method thereof. The Pyracantha fortuneana fruit extract prepared by the method of the present invention has significant anti-aging and whitening effects.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The invention provides a method for preparing an anti-aging and whitening Pyracantha fruit extract by using a supramolecular solvent, comprising: mixing and heating betaine, 1,3-propylene glycol and water to obtain a supramolecular solvent; drying and crushing Pyracantha fruit to obtain Pyracantha fruit powder, mixing the powder with the supramolecular solvent, extracting by hot reflux, filtering, standing, centrifuging a liquid, and obtaining a supernatant as the Pyracantha fruit extract; and the volume ratio of betaine to 1,3-propylene glycol is (1-5):(1-5).

[0008] Preferably, the solid-liquid ratio of the Pyracantha fruit powder to the supramolecular solvent is 1:5-50 g / mL, the temperature of the hot reflux extraction is 95-105° C., the time is 0.5-2 h, the centrifugal speed is 35000-4500 r / min, and the centrifugal time is 8-12 min.

[0009] The present invention also provides the Pyracantha fortuneana fruit extract prepared by the above method.

[0010] The present invention also provides a method for preparing an anti-aging and whitening Pyracantha fruit extract using a supramolecular solvent, which is characterized by comprising: mixing and heating betaine, 1,3-propylene glycol and water to obtain a supramolecular solvent; drying and crushing the Pyracantha fruit and mixing it with water, and sequentially subjecting it to composite enzyme hydrolysis and composite microbial fermentation, drying the fermentation liquid to obtain fermentation powder, ultrasonically extracting with the supramolecular solvent, centrifuging, and drying the supernatant to obtain the Pyracantha fruit extract; the volume ratio of betaine to 1,3-propylene glycol is (1-5):(1-5).

[0011] Preferably, the mass ratio of the Pyracantha fruit to water is 1:5-10.

[0012] Preferably, the complex enzyme is composed of pectate lyase and nattokinase in a weight ratio of 1-3:1-3, and the added amount of the complex enzyme is 3‰-8‰ of the mass of the Pyracantha fruit; the temperature of the enzymolysis is 40-50°C, the pH value is 7-8, and the time is 1-3h.

[0013] Preferably, the composite microorganism is composed of Bifidobacterium adolescentis and Lactobacillus johnsonii in a weight ratio of 2-5:1, and the added amount of the composite microorganism is 3%-8% of the weight of the Pyracantha fortuneana fruit; the fermentation temperature is 35-40°C, the pH value is 6.5-7.5, and the time is 2-5d.

[0014] Preferably, the solid-liquid ratio of the baking powder to the supramolecular solvent is 1:5-10 g / mL; the temperature of the supramolecular solvent ultrasonic extraction is 40-50° C., the power is 300-400 W, and the time is 20-30 min.

[0015] The present invention also provides the Pyracantha fortuneana fruit extract obtained by the above preparation method.

[0016] The present invention also provides the use of the pyracantha fruit extract in preparing anti-aging and whitening products.

[0017] Compared with the prior art, the present invention has the following beneficial effects: The present invention uses betaine as a hydrogen bond acceptor and 1,3-propylene glycol as a hydrogen bond donor, and mixes and self-assembles the two to form a supramolecular solvent; and uses the supramolecular solvent to extract Pyracantha fruit. The present invention uses a supramolecular solvent that is harmless to the skin to extract the active ingredients in Pyracantha fruit, thereby avoiding the problem of solvent irritation to the skin. The molecular solvent of the present invention can specifically bind to the active ingredients in Pyracantha fruit, thereby improving the purity and activity of the active ingredients. The Pyracantha fruit extract obtained by extraction with a supramolecular solvent has a good ability to scavenge DPPH free radicals and ABTS free radicals, and has a good effect on Fe 3+It has a high reducing ability; it has an inhibitory effect on the formation of non-enzymatic glycosylation intermediates - ketoamine compounds, and can be used for glycation anti-aging; it has an inhibitory effect on AAPH-induced oxidative damage to red blood cells, and has antioxidant and soothing effects; it has an inhibitory effect on elastase and has anti-aging effects; it can inhibit the activity of tyrosinase, has a whitening effect, can reduce facial pigmentation, and brighten the skin.

[0018] The combination of the pyracantha fruit extract and the hawk claw bean flower extract has a synergistic effect, can improve skin wrinkles and skin brightness, and has significant anti-aging and whitening effects. DETAILED DESCRIPTION

[0019] The invention provides a method for preparing an anti-aging and whitening Pyracantha fruit extract by using a supramolecular solvent, comprising: mixing and heating betaine, 1,3-propylene glycol and water to obtain a supramolecular solvent; drying and crushing Pyracantha fruit to obtain Pyracantha fruit powder, mixing the powder with the supramolecular solvent, extracting by hot reflux, filtering, standing, centrifuging the liquid, and obtaining the supernatant as the Pyracantha fruit extract; and the mass ratio of betaine to 1,3-propylene glycol is (1-5):(1-5).

[0020] The betaine of the present invention is a hydrogen bond acceptor, 1,3-propylene glycol is a hydrogen bond donor, and betaine and 1,3-propylene glycol can be self-assembled by heating in an aqueous solution to form a supramolecular solvent. The supramolecular solvent of the present invention is harmless to the skin, has a unique solubility to extract the effective components in the pyracantha fruit, and avoids the problem of solvent irritation to the skin.

[0021] The solid-liquid ratio of the Pyracantha fruit powder to the supramolecular solvent of the present invention is preferably 1:5-50 g / mL, more preferably 1:30 g / mL, the temperature of the hot reflux extraction is preferably 95-105°C, more preferably 100°C, the time is preferably 0.5-2h, more preferably 1.5h, the centrifugal speed is preferably 35000-4500r / min, more preferably 4000r / min, and the centrifugal time is preferably 8-12min, more preferably 10min.

[0022] The present invention also provides the Pyracantha fortuneana fruit extract prepared by the above method.

[0023] The present invention also provides a method for preparing an anti-aging and whitening Pyracantha fruit extract using a supramolecular solvent, comprising: mixing and heating betaine, 1,3-propylene glycol and water to obtain a supramolecular solvent; drying and crushing the Pyracantha fruit and mixing it with water, and subjecting the mixture to composite enzyme hydrolysis and composite microbial fermentation in sequence, drying the fermentation liquid to obtain fermentation powder, ultrasonically extracting with the supramolecular solvent, centrifuging, and drying the supernatant to obtain the Pyracantha fruit extract; the volume ratio of betaine to 1,3-propylene glycol is (1-5):(1-5).

[0024] The preparation method of the supramolecular solvent of the present invention preferably comprises: mixing betaine, 1,3-propylene glycol and water to obtain a mixture, heating and stirring in a water bath at 80° C. until a uniform liquid is formed in a beaker, cooling to obtain a supramolecular solvent, and storing in the dark; the water content in the mixture is 30%-90%.

[0025] The mass ratio of the Pyracantha fruit to water of the present invention is preferably 1:5-10, more preferably 1:8.

[0026] The composite enzyme of the present invention is preferably composed of pectate lyase and nattokinase in a weight ratio of 1-3:1-3, more preferably 2:2; the enzyme activity of the pectate lyase is preferably 100,000-200,000 U / g, more preferably 150,000 U / g, and the enzyme activity of the nattokinase is preferably 50,000-100,000 U / g, more preferably 80,000 U / g; the addition amount of the composite enzyme is preferably 3‰-8‰ of the mass of the pyracantha fruit, more preferably 5‰; the temperature of the enzymolysis is preferably 40-50°C, more preferably 45°C, the pH value is preferably 7-8, more preferably 7.5, and the time is preferably 1-3h, more preferably 2h. The composite enzymes of the present invention have a synergistic effect, pectate lyase destroys from the pectin level of the cell wall, and nattokinase destroys from the protein level of the intercellular matrix, more comprehensively and efficiently destroying the overall structure of the pyracantha fruit cells, making the effective ingredients in the cells easier to release, and can generate more active ingredients or enhance their activity, increasing the efficacy of the pyracantha fruit enzymolysis products.

[0027] The composite microorganism of the present invention is preferably composed of Bifidobacterium adolensentis and Lactobacillus johnsonii in a weight ratio of 2-5:1, more preferably 3:1. The Bifidobacterium adolensentis is Bifidobacterium adolensentis CICC ® 6178, purchased from China Industrial Microbiological Culture Collection Management Center, with a bacterial activity of 5×10 8 -8×10 9 CFU / g; the Lactobacillus johnsonii is Lactobacillus johnsonii MH-68, with a collection number of CCTCC NO: M2011128, purchased from China Center for Type Culture Collection, disclosed in CN112075638A, and a bacterial activity of 8×10 8 -5×10 9CFU / g. The addition amount of the composite microorganism is preferably 3%-8% of the weight of the pyracantha fruit, more preferably 5%; the fermentation temperature is preferably 35-40°C, more preferably 37°C, the pH value is preferably 6.5-7.5, more preferably 7, and the time is preferably 2-5d, more preferably 3d. The composite microorganisms of the present invention have a synergistic effect. The sugar decomposition of Bifidobacterium adolescentis provides a rich nutritional substrate for Lactobacillus johnsonii, and the metabolites produced by Lactobacillus johnsonii are conducive to the continuous growth of Bifidobacterium adolescentis and the maintenance of enzyme activity. The two cooperate with each other to make the fermentation process more efficient, and a good fermentation degree can be achieved in a shorter time. It also significantly increases the content and activity of the active ingredients in the pyracantha fruit extract, so that the pyracantha fruit extract may have more significant effects in anti-oxidation, anti-aging, whitening, etc., and enhances its application value in related fields.

[0028] The solid-liquid ratio of the baking powder and the supramolecular solvent of the present invention is 1:5-10g / mL, more preferably 1:8g / mL; the temperature of the ultrasonic extraction of the supramolecular solvent is preferably 40-50°C, more preferably 45°C, the power is preferably 300-400W, more preferably 350W, and the time is preferably 20-30min, more preferably 25min. The supramolecular solvent of the present invention can specifically bind to the active ingredients in the baking powder, thereby improving the purity and activity of polyphenols and flavonoids in the extract; ultrasonic treatment accelerates the mass transfer process between the solvent and the components in the baking powder, making the extraction process more efficient. The synergy of the two greatly improves the extraction efficiency and shortens the extraction time. Compared with the use of supramolecular solvents or ultrasound alone, a higher content of extract can be obtained in a shorter time.

[0029] The invention crushes the pyracantha fruit, which is beneficial to the dissolution of cell contents; the compound enzyme hydrolysis can not only destroy the cell wall structure, but also decompose the cell contents into small molecular substances, which is more beneficial to the fermentation of microorganisms; the compound microbial fermentation can further decompose the macromolecular substances in the pyracantha fruit and convert them into small molecular active components that are easier to extract; the supramolecular solvent can specifically combine with the active components in the fermentation powder to improve the purity and activity of the extract; the ultrasonic treatment accelerates the mass transfer process between the solvent and the components in the fermentation powder, making the extraction process more efficient.

[0030] The present invention also provides the Pyracantha fortuneana fruit extract obtained by the above preparation method.

[0031] The pyracantha fruit extract obtained by the present invention contains multiple active ingredients, such as flavonoids, polyphenols, vitamins, enzymes produced during the fermentation process, etc. These active ingredients have good antioxidant reduction ability; they have an inhibitory effect on the generation of non-enzymatic glycosylation intermediates, which is beneficial to glycation and anti-aging; they improve red blood cell oxidative damage and have an antioxidant and soothing effect; they have an inhibitory effect on elastase and tyrosinase, can resist aging and whiten, reduce facial pigmentation, and brighten the skin.

[0032] The present invention also provides application of the Pyracantha fortuneana fruit extract in preparing anti-aging and whitening products.

[0033] The present invention also provides an anti-aging and whitening composition containing the Pyracantha fortuneana fruit extract, comprising the following raw materials in parts by weight: 0.3-0.5 parts of Pyracantha fortuneana fruit extract, 1-5 parts of betaine, 5-10 parts of propylene glycol and 1-3 parts of 1,2-hexanediol.

[0034] The anti-aging and whitening composition of the present invention also includes hawkthorn bean flower extract and parahydroxyacetophenone. Hawkthorn bean flower extract was purchased from Zhongshan Honglai Biotechnology Co., Ltd. Pyracantha fruit extract is rich in flavonoids, polyphenols and other ingredients, which can inhibit tyrosinase activity. Hawkthorn bean flower extract can interfere with the signal transduction pathway in melanocytes, reduce the expression of melanin production-related genes, and reduce the synthesis of melanin. The two can achieve whitening effects from different aspects. Both Pyracantha fruit extract and hawkthorn bean flower extract contain antioxidant active ingredients, which can more effectively delay the cell aging process. Pyracantha fruit extract focuses on regulating signal pathways to start collagen synthesis, while hawkthorn bean flower extract plays a role in promoting cell proliferation and optimizing collagen secretion. The two jointly promote the synthesis and quality improvement of collagen, so that the elasticity and firmness of the skin are better maintained, thereby enhancing the anti-aging effect and reducing wrinkles and sagging.

[0035] The method of using the anti-aging and whitening composition of the present invention is as follows: after cleaning the face, take a proper amount, spray and rub it evenly on the face, pat gently until absorbed, and use it in the morning and evening.

[0036] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0038] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0039] Example 1 Preparation of Pyracantha fortuneana fruit extract (1) Preparation of supramolecular solvents betaine, 1,3-propylene glycol and water were mixed to obtain a mixture, which was heated and stirred in a water bath at 80° C. until a uniform liquid was formed in a beaker, and the mixture was cooled to obtain a supramolecular solvent, which was stored away from light; The mass ratio of betaine to 1,3-propylene glycol is 3:3; the volume percentage of water in the mixture is 50%; (2) Extraction of Pyracantha fruit The pyracantha fruit was dried at 45°C to a moisture content of 5%, and crushed through a 500-mesh sieve to obtain pyracantha fruit powder; the pyracantha fruit powder was mixed with a supramolecular solvent at a solid-liquid ratio of 1:30 g / mL, and extracted by hot reflux at 100°C for 1 hour. The extract was filtered and refrigerated at 4°C overnight, the upper solution was centrifuged at 4000 r / min for 10 minutes, and the supernatant was taken as the pyracantha fruit extract.

[0040] Test Example 1 Determination of Total Flavonoids in Pyracantha Fortuneana Fruit Extract After the pyracantha fruit extract prepared in Example 1 was diluted 4 times with PBS solution, 0.4 mL was taken out, and water (1.6 mL) and 5% sodium nitrite (0.12 mL) were added in turn. After incubation for 5 min, aluminum nitrate (0.12 mL) was added. After reaction for 6 min, 1 mol / L sodium hydroxide (0.8 mL) and water (0.96 mL) were added in turn. After reaction for 10 min, the absorbance was measured at 510 nm. The solvent was used as a blank control and rutin was used as a reference substance to draw a standard curve. The total flavonoid content was expressed as how many milligrams of rutin per gram of sample. The linear regression equation of the standard curve of total flavonoid content is: y=0.5943x-0.0089, R 2 =0.9998, which has a good linear correlation, so this equation can be used to calculate the total flavonoids content in the sample.

[0041] The total flavonoid content in the Pyracantha fortuneana fruit extract of Example 1 was calculated by the linear regression equation of the total flavonoid standard curve to be (1.698±0.021) mg / mL.

[0042] Example 2 Preparation of Pyracantha fortuneana fruit extract (1) Preparation of supramolecular solvents betaine, 1,3-propylene glycol and water were mixed to obtain a mixture, which was heated and stirred in a water bath at 80° C. until a uniform liquid was formed in a beaker, and the mixture was cooled to obtain a supramolecular solvent, which was stored away from light; The mass ratio of betaine to 1,3-propylene glycol is 3:3; the volume percentage of water in the mixture is 50%; (2) Pretreatment and enzymatic hydrolysis of Pyracantha fruit The pyracantha fruit was dried at 45°C to a moisture content of 5%, and crushed through a 500-mesh sieve to obtain pyracantha fruit powder; the pyracantha fruit powder was mixed with water at a weight ratio of 1:8, and a composite enzyme was added, and enzymolysis was performed at 45°C and a pH value of 7.5 for 2 hours, and the enzyme was inactivated at 85°C for 15 minutes to obtain an enzymolysis solution; The composite enzyme is preferably composed of pectate lyase and nattokinase in a weight ratio of 2:2, the pectate lyase activity is 150,000 U / g, the nattokinase activity is 80,000 U / g, and the added amount of the composite enzyme is 5‰ of the weight of the Pyracantha fortuneana fruit powder; (3) Fermentation Inoculating composite microorganisms into the enzymatic hydrolysate, fermenting at 37°C and pH 7 for 3 days, filtering out microbial cells, and spray-drying the fermentation liquid at 75°C to obtain baking powder; The composite microorganisms are composed of Bifidobacterium adolensentis and Lactobacillus johnsonii in a mass ratio of 3:1, and Bifidobacterium adolensentis is Bifidobacterium adolensentis CICC. ® 6178, bacterial activity is 5×10 9 CFU / g, Lactobacillus johnsonii is Lactobacillus johnsonii MH-68, the deposit number is CCTCC NO: M2011128, and the bacterial activity is 3×10 9 CFU / g, the inoculum amount of composite microorganisms was 5% of the weight of Pyracantha fortuneana fruit powder; (4) Ultrasonic extraction of supramolecular solvents The fermentation powder is mixed with the supramolecular solvent of step (1) at a solid-liquid ratio of 1:8 g / mL, ultrasonic extraction is performed at 45° C. and 350 W for 25 min, centrifugation is performed at 2000 rpm for 10 min, and the supernatant is taken to obtain the Pyracantha fortuneana fruit extract.

[0043] Example 3 Preparation of Pyracantha fortuneana fruit extract (1) Preparation of supramolecular solvents betaine, 1,3-propylene glycol and water were mixed to obtain a mixture, which was heated and stirred in a water bath at 75° C. until a uniform liquid was formed in a beaker, and the mixture was cooled to obtain a supramolecular solvent, which was stored away from light; The mass ratio of betaine to 1,3-propylene glycol is 1:5; the volume percentage of water in the mixture is 30%; (2) Pretreatment and enzymatic hydrolysis of Pyracantha fruit The pyracantha fruit was dried at 40°C to a moisture content of 7%, and crushed through a 400-mesh sieve to obtain pyracantha fruit powder; the pyracantha fruit powder was mixed with water at a weight ratio of 1:5, and a composite enzyme was added to perform enzymolysis at 40°C and pH 7 for 3 hours, and the enzyme was inactivated at 80°C for 20 minutes to obtain an enzymolysis solution; The composite enzyme is preferably composed of pectate lyase and nattokinase in a weight ratio of 1:3, the pectate lyase activity is 150,000 U / g, the nattokinase activity is 80,000 U / g, and the added amount of the composite enzyme is 3‰ of the weight of the Pyracantha fortuneana fruit powder; (3) Fermentation Inoculating composite microorganisms into the enzymatic hydrolysate, fermenting at 35°C and pH 6.5 for 5 days, filtering out the microbial cells, and spray-drying the fermentation liquid at 70°C to obtain baking powder; The composite microorganisms are composed of Bifidobacterium adolensentis and Lactobacillus johnsonii in a mass ratio of 2:1, and Bifidobacterium adolensentis is Bifidobacterium adolensentis CICC. ® 6178, bacterial activity is 5×10 8 CFU / g, Lactobacillus johnsonii is Lactobacillus johnsonii MH-68, the preservation number is CCTCC NO: M2011128, and the bacterial activity is 8×10 8 The inoculum amount of CFU / g composite microorganisms was 3% of the weight of Pyracantha fruit powder; (4) Ultrasonic extraction of supramolecular solvents The fermentation powder is mixed with the supramolecular solvent of step (1) at a solid-liquid ratio of 1:5 g / mL, ultrasonically extracted at 40° C. and 300 W for 30 min, centrifuged at 1500 rpm for 20 min, and the supernatant is collected to obtain the Pyracantha fortuneana fruit extract.

[0044] Example 4 Preparation of Pyracantha fortuneana fruit extract (1) Preparation of supramolecular solvents betaine, 1,3-propylene glycol and water were mixed to obtain a mixture, which was heated and stirred in a water bath at 85° C. until a uniform liquid was formed in a beaker, and the mixture was cooled to obtain a supramolecular solvent, which was stored away from light; The mass ratio of betaine to 1,3-propylene glycol is 5:1; the volume percentage of water in the mixture is 90%; (2) Pretreatment and enzymatic hydrolysis of Pyracantha fruit The pyracantha fruit was dried at 50°C to a moisture content of 3%, and crushed through a 600-mesh sieve to obtain pyracantha fruit powder; the pyracantha fruit powder was mixed with water at a weight ratio of 1:10, and a composite enzyme was added, and enzymolysis was performed at 50°C and pH 8 for 1 hour, and the enzyme was inactivated at 90°C for 10 minutes to obtain an enzymolysis solution; The composite enzyme is preferably composed of pectate lyase and nattokinase in a weight ratio of 3:1, the pectate lyase activity is 150,000 U / g, the nattokinase activity is 80,000 U / g, and the added amount of the composite enzyme is 8‰ of the weight of the Pyracantha fortuneana fruit powder; (3) Fermentation Inoculating composite microorganisms into the enzymatic hydrolysate, fermenting at 40°C and pH 7.5 for 2 days, filtering out microbial cells, and spray-drying the fermented liquid at 80°C to obtain baking powder; The composite microorganisms are composed of Bifidobacterium adolensentis and Lactobacillus johnsonii in a mass ratio of 5:1. Bifidobacterium adolensentis CICC ® 6178, bacterial activity is 8×10 9 CFU / g, Lactobacillus johnsonii is Lactobacillus johnsonii MH-68, the preservation number is CCTCC NO: M2011128, and the bacterial activity is 5×10 9 CFU / g, the inoculum amount of composite microorganisms was 5% of the weight of Pyracantha fortuneana fruit powder; (4) Ultrasonic extraction of supramolecular solvents The fermentation powder is mixed with the supramolecular solvent of step (1) at a solid-liquid ratio of 1:10 g / mL, ultrasonically extracted at 50° C. and 400 W for 20 min, centrifuged at 2000 rpm for 8 min, and the supernatant is collected to obtain the Pyracantha fortuneana fruit extract.

[0045] Comparative Example 1 The specific implementation method is the same as Example 2, except that the composite enzyme is composed of cellulase and snail protease, preferably in a weight ratio of 2:2, the cellulase activity is 150,000 U / g, the snail protease activity is 80,000 U / g, the added amount of the composite enzyme is 5‰ of the weight of the Pyracantha fruit powder, and the enzymatic hydrolysis conditions are 45°C, pH 7.5, and time is 2h.

[0046] Comparative Example 2 The specific implementation method is the same as that of Example 2, except that the composite microorganism is composed of Bifidobacterium longum and Lactobacillus plantarum in a mass ratio of 3:1, and Bifidobacterium longum is Bifidobacterium longum CICC ® 6209, bacterial activity is 5×10 9 CFU / g, Lactobacillus plantarum CICC ® 24936, bacterial activity is 3×10 9 CFU / g, the inoculation amount of the composite microorganisms was 5% of the weight of the Pyracantha fruit powder, and the fermentation parameters were 37°C, pH 7, and time for 3 days.

[0047] Comparative Example 3 The specific implementation method is the same as that of Example 2, except that the mass ratio of betaine to 1,3-propylene glycol in the supramolecular solvent is 1:7.

[0048] Comparative Example 4 The specific implementation method is the same as that of Example 2, except that the mass ratio of betaine to 1,3-propylene glycol in the supramolecular solvent is 7:1.

[0049] Comparative Example 5 The specific implementation is the same as that of Example 2, except that 1,3-propanediol in the supramolecular solvent is replaced by n-butanol.

[0050] Test Example 2 Performance Test of Pyracantha Fruit Extract The test Pyracantha fortuneana fruit extract solution was diluted with PBS solution.

[0051] Take 1% Pyracantha fortuneana fruit extract solution as an example: mix the Pyracantha fortuneana fruit extract and PBS solution evenly, wherein the volume ratio of the Pyracantha fortuneana fruit extract to the PBS solution is 1:99.

[0052] (1) Antioxidant properties The DPPH free radical scavenging ability of the Pyracantha fortuneana fruit extract solutions of Examples 2-4 and Comparative Examples 1-5 at different volume concentrations of 0.5%, 1% and 3% was tested, with 1 mg / mL vitamin C as a positive control.

[0053] The ABTS free radical scavenging ability of the Pyracantha fortuneana fruit extract solutions of Examples 2-4 and Comparative Examples 1-5 at different volume concentrations of 2%, 5% and 10% was tested, with 1 mg / mL vitamin C as a positive control.

[0054] The effects of different volume concentrations of 2%, 5% and 10% of the extracts of Example 2-4 and Comparative Example 1-5 on Fe in 1 mM / L ferric chloride solution were detected. 3+ restoration capability.

[0055] The experiments were repeated three times. The specific results of DPPH free radical scavenging ability, ABTS free radical scavenging ability and iron ion reducing ability of different Pyracantha fortuneana fruit extracts are shown in Table 1.

[0056] Table 1 Effects of different Pyracantha fruit extracts on DPPH free radical scavenging rate (%)

[0057] The scavenging rate of DPPH free radicals by 1 mg / mL vitamin C was 90.02%.

[0058] Table 2 Effects of different Pyracantha fruit extracts on ABTS free radical scavenging rate (%)

[0059] The scavenging rate of 1 mg / mL vitamin C on ABTS free radicals was 96.32%.

[0060] Table 3 Fe in different Pyracantha fruit extracts 3+ The impact of reduction ability

[0061] From the data in Tables 1 to 3, it can be seen that different Pyracantha fruit extracts have different degrees of inhibitory effects on DPPH free radicals and ABTS free radicals, and on Fe 3+ The reducing abilities are different. It can be seen from the data of Example 2 and Comparative Examples 1-2 that different composite enzymes and different composite microorganisms have an effect on the antioxidant properties of the Pyracantha fruit extract. It can be seen from the data of Example 2 and Comparative Examples 3-5 that the ratios of different betaine and 1,3-propylene glycol in the supramolecular solvent and the types of different supramolecular solvents have an effect on the antioxidant properties of the Pyracantha fruit extract. The Pyracantha fruit extract prepared by the preparation method of the present invention has excellent free radical scavenging ability, that is, it has excellent antioxidant ability.

[0062] (2) Inhibition of non-enzymatic glycosylation Non-enzymatic glycosylation (also known as glycation reaction) is a reaction between the aldehyde or ketone groups of sugars and the free amino groups of macromolecules such as proteins, lipids or nucleic acids without the participation of enzymes, forming irreversible advanced glycation end products. Non-enzymatic glycosylation is one of the important factors leading to skin aging. The determination of inhibition rate can directly reflect the degree of inhibition of the active ingredients in cosmetics on the non-enzymatic glycosylation process, providing a quantitative scientific basis for the anti-glycation effect claimed by the product.

[0063] For the specific experimental process, please refer to "Inhibitory effect of Bifidobacterium longum extracellular polysaccharides on non-enzymatic glycosylation" (Xia Junying, Yu Haixia, Qi Shimei, et al. Inhibitory effect of Bifidobacterium longum extracellular polysaccharides on non-enzymatic glycosylation [J]. Journal of Biology, 2024, 41(05): 6-13+47) 1.2.2 Establishment of MGO+BSA non-enzymatic glycosylation model and detection of characteristic products (2) Determination method of dicarbonyl compound content.

[0064] The content of dicarbonyl compounds in the Pyracantha fortuneana fruit extract solutions of different volume concentrations of 10% and 20% were detected, and the inhibition rate was calculated. PBS solution was used as a blank control, and 1 mg / ml aminoguanidine hydrochloride was used as a positive control. The specific results are shown in Table 4.

[0065] Table 4 Effects of different Pyracantha fruit extracts on inhibition rate of dicarbonyl compounds (%)

[0066] The inhibition rate of dicarbonyl compounds of 1 mg / ml aminoguanidine hydrochloride was 66.93%.

[0067] It can be seen from the data in Table 4 that Examples 2-4 of the present invention have the strongest inhibition rate on dicarbonyl compounds. It can be seen from the data in Example 2 and Comparative Examples 1-2 that different composite enzymes and different composite microorganisms have an effect on the inhibition rate of dicarbonyl compounds in Pyracantha fruit extract. It can be seen from the data in Example 2 and Comparative Examples 3-5 that the ratios of different betaine and 1,3-propylene glycol in the supramolecular solvent and the types of different supramolecular solvents have an effect on the inhibition rate of dicarbonyl compounds in Pyracantha fruit extract. The Pyracantha fruit extract prepared by the preparation method of the present invention has an inhibitory effect on saccharification reaction and can be used for saccharification and anti-aging.

[0068] (3) AAPH-induced erythrocyte hemolysis rate and hemolysis inhibition rate AAPH is a free radical initiator commonly used in the erythrocyte oxidative hemolysis model. At 37°C, AAPH can decompose to produce alkyl free radicals, which can be oxidized into hydroperoxyl free radicals. Hydroperoxyl free radicals react with phospholipids on the phospholipid bilayer on the cell membrane, causing lipid peroxidation, thereby destroying the cell membrane and causing cell hemolysis. The AAPH-induced erythrocyte hemolysis model was used to explore the antioxidant activity of the samples at the cellular level and their potential mechanisms of action.

[0069] The specific experimental process refers to "Study on Ultrasonic-Assisted Enzymatic Extraction of Mulberry Leaf Flavonoids and Their Antioxidant Activity" (Zheng Qinhua. Study on Ultrasonic-Assisted Enzymatic Extraction of Mulberry Leaf Flavonoids and Their Antioxidant Activity [D]. Jilin University, 2023. DOI: 10.27162 / d.cnki.gjlin.2023.006866.) The experimental process of 4.3.5-4.3.6 was used to detect the inhibition rate of 2%, 5% and 10% of the pyracantha fruit extract solutions of Examples 1-3 and Comparative Examples 1-5 on oxidative damage and hemolysis of red blood cells. PBS solution was used as a blank control, and AAPH oxidatively damaged cells were used as a positive control. The specific results are shown in Table 5.

[0070] Table 5 Effects of different Pyracantha fortuneana fruit extracts on the inhibition rate of erythrocyte oxidative damage hemolysis (%)

[0071] The inhibition rate of AAPH oxidative damage cells on erythrocyte oxidative damage hemolysis was 48.43%, and the inhibition rate of PBS blank control erythrocyte oxidative damage hemolysis was 83.26%.

[0072] It can be seen from the data in Table 5 that the inhibition rate of hemolysis of oxidative damage to red blood cells in Examples 2-4 of the present invention is stronger than that in the comparative example, which also indicates that the hemolysis irritation to red blood cells is lower. It can be seen from the data in Example 2 and Comparative Examples 1-2 that different composite enzymes and different composite microorganisms have an effect on the inhibition rate and irritation of hemolysis of oxidative damage to red blood cells of the Pyracantha fruit extract. It can be seen from the data in Example 2 and Comparative Examples 3-5 that the ratios of different betaine and 1,3-propylene glycol in the supramolecular solvent and the types of different supramolecular solvents have an effect on the inhibition rate and irritation of hemolysis of oxidative damage to red blood cells of the Pyracantha fruit extract. The Pyracantha fruit extract prepared by the preparation method of the present invention can scavenge alkyl free radicals, protect cells from damage, and has antioxidant and soothing effects.

[0073] (4) Elastase inhibition rate Elastase is one of the most important enzymes in the matrix metalloproteinase family. It is mainly synthesized and secreted by fibroblasts. It can degrade elastin in the skin and activate the activity of other matrix metalloproteinases, accelerating the degradation of the extracellular matrix in the dermis, thus causing serious damage to the normal structure and function of the dermis of human skin, causing obvious wrinkles and loss of elasticity. Therefore, inhibiting the activity of elastase can slow down skin aging to a certain extent.

[0074] The specific experimental process refers to the elastase inhibition test in paragraph

[0030] of the invention patent CN 103385917 A to determine the inhibition rates of the elastase by the pyracantha fruit extract solutions of Examples 2-4 and Comparative Examples 1-5 at different volume concentrations of 1%, 5% and 10%. The specific results are shown in Table 6.

[0075] Table 6 Inhibition rate of different Pyracantha fortuneana fruit extracts on elastase (%)

[0076] It can be seen from the data in Table 6 that as the concentration of the Pyracantha fruit extract solution increases, the inhibition rate of elastase increases, and the Pyracantha fruit extract solutions of Examples 2-4 of the present invention have higher inhibition ability on elastase than Comparative Examples 1-5. It can be seen from the data of Example 2 and Comparative Examples 1-2 that different composite enzymes and different composite microorganisms have an effect on the elastase inhibition rate of the Pyracantha fruit extract. It can be seen from the data of Example 2 and Comparative Examples 3-5 that the ratios of different betaines and 1,3-propylene glycol in the supramolecular solvent and the types of different supramolecular solvents have an effect on the elastase inhibition ability of the dicarbonyl compounds of the Pyracantha fruit extract. The Pyracantha fruit extract prepared by the preparation method of the present invention has good anti-aging effect.

[0077] (5) Tyrosinase inhibition rate Tyrosinase is a key enzyme in melanin anabolism and plays an important role in the melanin synthesis process. The activity of tyrosinase directly determines the amount of melanin formed.

[0078] The specific experimental process refers to the detection method of half inhibition rate in paragraph [0020-0023] of invention patent CN 107822996 A, and the half inhibition rate (IC50) concentration of the Pyracantha fortuneana fruit extracts of Examples 2-4 and Comparative Examples 1-5 is measured. The specific results are shown in Table 7, where the half inhibition rate is the mass concentration.

[0079] Table 7 Half inhibition rate of tyrosinase of different Pyracantha fortuneana fruit extracts (%)

[0080] It can be seen from the data in Table 7 that the half inhibition rate of tyrosinase in the Pyracantha fruit extract of the present invention is lower than that in the mass concentration of Comparative Examples 1-5. It can be seen from the data in Example 2 and Comparative Examples 1-2 that different composite enzymes and different composite microorganisms have an effect on the elastase inhibition rate of the Pyracantha fruit extract. It can be seen from the data in Example 2 and Comparative Examples 3-5 that the ratio of different betaine and 1,3-propylene glycol in the supramolecular solvent and the types of different supramolecular solvents have an effect on the mass concentration of the half inhibition rate of the Pyracantha fruit extract. The Pyracantha fruit extract prepared by the preparation method of the present invention has a good whitening effect, can reduce facial pigmentation, and brighten the skin.

[0081] Example 5 Preparation of anti-aging and whitening composition of pyracantha fruit extract Weigh 0.4 parts by weight of the pyracantha fruit extract of Example 2, 3 parts by weight of the hawkthorn bean flower extract, 3 parts by weight of betaine, 8 parts by weight of propylene glycol, 2 parts by weight of 1,2-hexanediol, 0.3 parts by weight of p-hydroxyacetophenone and 83.3 parts by weight of deionized water; Deionized water is placed in a stirring container, the speed is set to 300 r / min, betaine is placed in the deionized water until the betaine is completely dissolved to obtain a betaine solution; after propylene glycol and 1,2-hexanediol are mixed, hydroxyacetophenone is slowly added at a speed of 300 r / min until the hydroxyacetophenone is dissolved, the mixture is poured into the betaine solution, and stirred at a speed of 300 r / min for 15 minutes to obtain a mixed solution; pyracantha fruit extract and hawkthorn bean flower extract are slowly added to the mixed solution, homogenized at 3000 r / min for 10 minutes, and sterilized to obtain an anti-aging and whitening composition.

[0082] Example 6 Anti-aging and whitening composition of pyracantha fruit extract Weigh 0.5 parts by weight of the pyracantha fruit extract of Example 3, 2 parts by weight of the hawk claw bean flower extract, 1 part by weight of betaine, 6 parts by weight of propylene glycol, 1 part by weight of 1,2-hexanediol, 0.1 parts by weight of p-hydroxyacetophenone and 89.4 parts by weight of deionized water; Deionized water is put into a stirring container, the speed is set to 200 r / min, betaine is put into the deionized water until the betaine is completely dissolved to obtain a betaine solution; after propylene glycol and 1,2-hexanediol are mixed, hydroxyacetophenone is slowly added at a speed of 200 r / min until the hydroxyacetophenone is dissolved, and the mixture is poured into the betaine solution, and stirred at a speed of 200 r / min for 20 minutes to obtain a mixed solution; Pyracantha fruit extract and hawkthorn bean flower extract are slowly added to the mixed solution, homogenized at 2000 r / min for 20 minutes, and sterilized to obtain an anti-aging and whitening composition.

[0083] Example 7 Anti-aging and whitening composition of pyracantha fruit extract Weigh 0.3 parts by weight of the pyracantha fruit extract of Example 4, 4 parts by weight of the hawk claw bean flower extract, 5 parts by weight of betaine, 10 parts by weight of propylene glycol, 3 parts by weight of 1,2-hexanediol, 0.4 parts by weight of p-hydroxyacetophenone and 76.1 parts by weight of deionized water; Deionized water is placed in a stirring container, the speed is set to 400 r / min, betaine is placed in the deionized water until the betaine is completely dissolved to obtain a betaine solution; after propylene glycol and 1,2-hexanediol are mixed, hydroxyacetophenone is slowly added at a speed of 400 r / min until the hydroxyacetophenone is dissolved, and the mixture is poured into the betaine solution, and stirred at a speed of 400 r / min for 10 minutes to obtain a mixed solution; pyracantha fruit extract and hawkthorn bean flower extract are slowly added to the mixed solution, homogenized at 4000 r / min for 6 minutes, and sterilized to obtain an anti-aging and whitening composition.

[0084] Comparative Example 6 The specific implementation is the same as that of Example 5, except that the Pyracantha fortuneana fruit extract is removed and the weight portion of the pyracantha flower extract is 3.4 weight portions.

[0085] Comparative Example 7 The specific implementation is the same as that of Example 5, except that the hawkthorn bean flower extract is removed and the weight portion of the pyracantha fruit extract is 3.4 weight portions.

[0086] Comparative Example 8 The specific implementation is the same as that of Example 5, except that the Pyracantha fortuneana fruit extract is the Pyracantha fortuneana fruit extract prepared in Comparative Example 1.

[0087] Comparative Example 9 The specific implementation is the same as that of Example 5, except that the Pyracantha fortuneana fruit extract is the Pyracantha fortuneana fruit extract prepared in Comparative Example 2.

[0088] Comparative Example 10 The specific implementation is the same as that of Example 5, except that the Pyracantha fortuneana fruit extract is the Pyracantha fortuneana fruit extract prepared in Comparative Example 3.

[0089] Comparative Example 11 The specific implementation is the same as that of Example 5, except that the Pyracantha fortuneana fruit extract is the Pyracantha fortuneana fruit extract prepared in Comparative Example 4.

[0090] Comparative Example 12 The specific implementation is the same as that of Example 5, except that the Pyracantha fortuneana fruit extract is the Pyracantha fortuneana fruit extract prepared in Comparative Example 5.

[0091] Test Example 3 The subjects were aged 35-50 years old, with a male-female ratio of 1:1, without chronic diseases, major diseases and allergic constitution, and did not take any medicine. The subjects were divided into 10 groups, 30 people in each group, and there was no significant difference between each group. The anti-aging and whitening compositions of Examples 5-7 and Comparative Examples 6-12 were used respectively. The method of using the anti-aging and whitening composition is: after cleaning the face, take an appropriate amount of cleansing, spray and rub it evenly on the face, pat until absorbed, use it in the morning and evening, and use it continuously for 8 weeks. The skin brightness and skin elasticity of each group at the forehead were detected on the 0th day, the 28th day and the 56th day, and the skin brightness improvement rate and wrinkle improvement rate on the 28th day and the 56th day were calculated. The specific results are shown in Table 8.

[0092] After using the anti-aging and whitening composition for 30 minutes, the skin brightness and skin elasticity were measured using a skin analyzer in an environment with a temperature of 25±0.5°C and a relative humidity of 50±1%.

[0093] Skin brightness improvement rate (%) = (skin brightness on day n - skin brightness on day 0) ÷ skin brightness on day 0 × 100%; Skin wrinkle improvement rate (%) = (skin elasticity on day n - skin elasticity on day 0) ÷ skin elasticity on day 0 × 100%.

[0094] Table 8 Skin brightness improvement rate and wrinkle improvement rate of different anti-aging and whitening compositions (%)

[0095] It can be seen from the data in Table 8 that the skin brightness improvement rate and wrinkle improvement rate of the anti-aging and whitening composition of the present invention are the best, and it has excellent anti-aging and whitening effects. From the comparison of the data of Example 5 and Comparative Examples 6-7, it can be seen that the pyracantha fruit extract and the hawkthorn bean flower extract are used synergistically to improve the anti-aging and whitening effects. From the comparison of the data of Example 5 and Comparative Examples 8-12, it can be seen that different complex enzymes, complex microorganisms, and different supramolecular solvent types and proportions have an effect on the anti-aging and whitening effects of the composition. The pyracantha fruit extract obtained by the preparation method of the present invention combined with the hawkthorn bean flower extract can give the composition excellent anti-aging and whitening effects.

[0096] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing an anti-aging and whitening Pyracantha fortuneana fruit extract using a supramolecular solvent, characterized in that: include: Betaine, 1,3-propylene glycol and water are mixed and heated to obtain a supramolecular solvent; Pyracantha fruit is dried and crushed to obtain Pyracantha fruit powder, which is mixed with the supramolecular solvent, subjected to hot reflux extraction, filtered, allowed to stand, and the liquid is centrifuged to obtain the supernatant, which is the Pyracantha fruit extract; the volume ratio of betaine to 1,3-propylene glycol is (1-5):(1-5).

2. The method according to claim 1, characterized in that The solid-liquid ratio of the pyracantha fruit powder to the supramolecular solvent is 1:5-50 g / mL, the temperature of the hot reflux extraction is 95-105° C., the time is 0.5-2 h, the centrifugal speed is 35000-4500 r / min, and the centrifugal time is 8-12 min.

3. The Pyracantha fortuneana fruit extract prepared by the method of claim 1 or 2.

4. A method for preparing an anti-aging and whitening Pyracantha fortuneana fruit extract using a supramolecular solvent, characterized in that: include: Betaine, 1,3-propylene glycol and water are mixed and heated to obtain a supramolecular solvent; Pyracantha fruit is dried, crushed and mixed with water, and then subjected to composite enzyme hydrolysis and composite microbial fermentation in sequence, and the fermentation liquid is dried to obtain baking powder, and ultrasonic extraction is performed with the supramolecular solvent, and after centrifugation, the supernatant is dried to obtain the Pyracantha fruit extract; the volume ratio of betaine and 1,3-propylene glycol is (1-5):(1-5).

5. The method according to claim 4, characterized in that The mass ratio of the pyracantha fruit to water is 1:5-10.

6. The method according to claim 5, characterized in that The complex enzyme is composed of pectate lyase and nattokinase in a weight ratio of 1-3:1-3, and the added amount of the complex enzyme is 3‰-8‰ of the mass of the pyracantha fruit; the temperature of the enzymolysis is 40-50°C, the pH value is 7-8, and the time is 1-3h.

7. The method according to claim 4, characterized in that The composite microorganism is composed of Bifidobacterium adolescentis and Lactobacillus johnsonii in a weight ratio of 2-5:1, and the added amount of the composite microorganism is 3%-8% of the weight of the pyracantha fruit; the fermentation temperature is 35-40°C, the pH value is 6.5-7.5, and the fermentation time is 2-5 days.

8. The method according to claim 4, characterized in that The solid-liquid ratio of the baking powder to the supramolecular solvent is 1:5-10 g / mL; the temperature of the supramolecular solvent ultrasonic extraction is 40-50° C., the power is 300-400 W, and the time is 20-30 min.

9. The Pyracantha fortuneana fruit extract obtained by the preparation method according to any one of claims 4 to 8.

10. Use of the Pyracantha fortuneana fruit extract according to claim 3 or the Pyracantha fortuneana fruit extract according to claim 9 in the preparation of anti-aging and whitening products.

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