A whitening antioxidant composition base and its preparation method and application

Abstract

The present application relates to a kind of whitening antioxidant composition base and its preparation method and application, belong to antioxidant technical field.The whitening antioxidant composition base includes the following raw materials according to mass fraction: 20-30 parts of antarctic krill peptide, 1-10 parts of perilla seed peptide, 0.5-10 parts of flaxseed peptide, 1-15 parts of acai berry peptide, 0.05-5 parts of dendrobium extract, 15-25 parts of tea flower ferment.The present application is compounded by antarctic krill peptide, perilla seed peptide, flaxseed peptide, acai berry peptide, dendrobium extract and tea flower ferment, so that the composition base of the present application has excellent antioxidant property, reduces melanin, and inhibits tyrosinase function.

Description

Technical Field

[0001] This invention belongs to the field of antioxidant technology, specifically relating to a whitening and antioxidant composition base material, its preparation method, and its application. Background Technology

[0002] The skin, the largest organ in the human body, plays vital roles in barrier protection, thermoregulation, immune defense, sensory perception, and excretion. Skin aging is caused by a variety of internal and external factors, primarily manifesting as loss of skin elasticity, roughness, wrinkles, dullness, and pigmentation. Ultraviolet radiation is a major external factor contributing to skin aging, not only inducing the production of large amounts of reactive oxygen species (ROS) and accelerating the aging process, but also promoting melanocyte proliferation and stimulating melanin production. Melanin is synthesized within melanocytes, requiring several basic substances: tyrosine, tyrosinase, oxygen, and dopa. Tyrosine is the main raw material for melanin production, tyrosinase plays a crucial role in converting tyrosine into melanin, and oxygen is essential for dopa oxidation. With continuous social development and progress, people's quality of life has gradually improved, and skincare products have become increasingly common in households, with a growing demand for skincare products that offer whitening and anti-free radical benefits. Currently, whitening products on the market mainly contain chemically synthesized substances with whitening and antioxidant effects, such as arbutin, hydroquinone, kojic acid, BHT, vitamin C and its derivatives. Although these chemically synthesized compounds have significant effects, they have gradually revealed problems such as high irritation, instability and single function during use. Therefore, the development of safe, gentle, stable and multifunctional skin care products has become a hot topic of concern for research institutions and manufacturers in recent years. Summary of the Invention

[0003] The first objective of this invention is to provide a whitening and antioxidant composition base material to solve the technical problems of chemically synthesized substances being highly irritating and having limited effects.

[0004] The second objective of this invention is to provide a method for preparing a whitening and antioxidant composition base.

[0005] A third objective of this invention is to provide an application of a whitening and antioxidant composition base material.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] A whitening and antioxidant composition base material, calculated by weight, includes the following raw materials: 20-30 parts Antarctic krill peptide, 1-10 parts perilla seed peptide, 0.5-10 parts flaxseed peptide, 1-15 parts sacha indica peptide, 0.05-5 parts Dendrobium officinale extract, and 15-25 parts tea tree flower ferment.

[0008] Furthermore, the preparation method of the tea flower ferment includes the following steps:

[0009] S1: Add sterile water to tea tree pollen, sterilize and cool to obtain a primary fermentation substrate; inoculate the primary fermentation substrate with Schizophyllum commune for primary fermentation to obtain a primary fermentation product;

[0010] S2: After sterilizing and cooling the primary fermentation material, a secondary fermentation substrate is obtained. A mixture of yeast and Bacillus subtilis is introduced into the secondary fermentation substrate for secondary fermentation to obtain a secondary fermentation product. After sterilizing, centrifuging, and drying the secondary fermentation product, tea flower fermentation product is obtained.

[0011] Further, in S1, 20-50 mL of sterile water is added for every gram of tea pollen; the pH of the primary fermentation substrate is adjusted to 5.5-6.5 before inoculation with the *Schizophyllum commune* for fermentation; during the primary fermentation, the amount of *Schizophyllum commune* inoculated accounts for 5-8% of the volume of the primary fermentation substrate; the fermentation temperature for the primary fermentation is 25-30 ℃, the time is 3-5 days, and the rotation speed is 150-180 rpm; the viable count of the *Schizophyllum commune* is 1×10⁻⁶. 8 -10 10 CFU / g.

[0012] Furthermore, in step S2, the amount of the mixed yeast and Bacillus subtilis inoculated is 5-10% of the volume of the secondary fermentation substrate, and the viable count of the yeast is 5 × 10⁻⁶. 7 -5×10 9 CFU / g, the viable count of the Bacillus subtilis is 5 × 10⁻⁶. 7 -5×10 9 CFU / g; the mass ratio of yeast to Bacillus subtilis is 1:2-4; the temperature of the secondary fermentation is 28-35 ℃, the time is 24-72 h, and the rotation speed is 100-300 rpm.

[0013] Furthermore, the sterilization temperature in S1 and S2 is 110-121 ℃, and the sterilization time is 15-20 min; the primary fermentation substrate is obtained by cooling S1 to 25-30 ℃, and the secondary fermentation substrate is obtained by cooling S2 to 28-35 ℃.

[0014] Furthermore, Dendrobium officinale extracts can be prepared by water extraction, alcohol extraction, or other methods; the Dendrobium officinale extracts obtained by water extraction and alcohol extraction can be used in the cosmetics field as well as the food field.

[0015] In the cosmetics field, the preparation method of the Dendrobium officinale extract includes the following steps: adding Dendrobium officinale powder to an extraction solution, ultrasonically extracting to obtain a crude extract, centrifuging the crude extract and taking the supernatant, heating the supernatant, and drying the upper layer solution to obtain the final product; the extraction solution is an aqueous solution of a eutectic solvent, and the mass fraction of the eutectic solvent in the extraction solution is 60-70%; the eutectic solvent is composed of capric acid and tetracaine in a molar ratio of 1-2:1.

[0016] Furthermore, 20-30 mL of extraction solution is added for each gram of Dendrobium officinale powder; the ultrasonic extraction power is 50-100 W, the ultrasonic extraction temperature is 70-80 ℃, and the ultrasonic extraction time is 30-60 min; the supernatant is heated to a temperature of 60-70 ℃.

[0017] Furthermore, the Antarctic krill peptides contain >90% peptides with a molecular weight ≤5000 Da, ≥75% peptide content, and ≥80% protein content; the perilla seed peptides contain >80% peptides with a molecular weight ≤10000 Da, ≥40% peptide content, and ≥50% protein content; the flaxseed peptides contain >80% peptides with a molecular weight ≤10000 Da, ≥50% peptide content, and ≥60% protein content; and the sacha inchi peptides contain >85% peptides with a molecular weight ≤3000 Da, ≥65% peptide content, and ≥75% protein content.

[0018] A method for preparing a whitening and antioxidant composition base includes the following steps: mixing 50% by weight of Antarctic krill peptide, all by weight of Sacha indica peptide, all by weight of Perilla frutescens seed peptide, all by weight of flaxseed peptide, all by weight of Dendrobium officinale extract, all by weight of tea tree flower ferment, and the remaining 50% of Antarctic krill peptide in sequence, and mixing at 50-60 r / min for 20-30 min to obtain the product.

[0019] Application of a whitening and antioxidant composition base material in the preparation of whitening and antioxidant products.

[0020] The beneficial effects of this invention are:

[0021] Antarctic krill peptides, perilla seed peptides, flaxseed peptides, and sacha indica peptides possess excellent free radical scavenging abilities and superior antioxidant properties, reducing oxidative stress damage to the skin and minimizing UV damage. Dendrobium officinale polysaccharides have powerful free radical scavenging effects; the Dendrobium officinale extract prepared in this invention contains over 55% Dendrobium officinale polysaccharides, significantly enhancing its free radical scavenging ability. This invention, through the fermentation of tea tree flowers, enriches the variety of active ingredients in the tea tree flowers, increases the tyrosinase activity inhibition rate of the tea tree flowers, and significantly enhances the whitening efficacy of the whitening and antioxidant composition base of this invention. This invention, through the compounding of Antarctic krill peptides, perilla seed peptides, flaxseed peptides, sacha indica peptides, Dendrobium officinale extract, and tea tree flower fermentation products, gives the composition base of this invention excellent antioxidant properties, melanin reduction, and tyrosinase inhibition functions. The whitening and antioxidant composition base of this invention can be used not only in skincare products but also in antioxidant foods.

[0022] This invention discovered that tea flower fermentation, which first uses Schizophyllum commune for fermentation and then uses yeast and Bacillus subtilis for co-fermentation, has the optimal tyrosinase activity inhibition rate. Through single-factor experiments, it was found that when the amount of Schizophyllum commune inoculated was 7% (v / v), the amount of yeast and Bacillus subtilis mixed inoculated was 8% (v / v), and the weight ratio of yeast to Bacillus subtilis was 1:4, the tyrosinase activity inhibition rate of the tea flower fermentation could reach as high as 61.35%, which significantly improved the whitening effect of tea flower. Detailed Implementation

[0023] The following will provide further details with reference to embodiments of the present invention.

[0024] In this invention, the Antarctic krill peptides contain >90% peptides with a molecular weight ≤5000 Da, ≥75% peptide content, and ≥80% protein content; the perilla seed peptides contain >80% peptides with a molecular weight ≤10000 Da, ≥40% peptide content, and ≥50% protein content; the flaxseed peptides contain >80% peptides with a molecular weight ≤10000 Da, ≥50% peptide content, and ≥60% protein content; and the sacha inchi peptides contain >85% peptides with a molecular weight ≤3000 Da, ≥65% peptide content, and ≥75% protein content.

[0025] The preparation method of Antarctic krill peptides includes: heading and shelling fresh Antarctic krill, followed by mincing using a meat grinder to obtain minced meat. Water is added at a ratio of 1 g: 10-20 mL to form a homogenate. The mixture is then hydrolyzed with protease at 55 ℃ for 3-6 h, followed by enzyme inactivation at 90-95 ℃ for 20 min. The mixture is filtered through a plate and frame filter to obtain a filtrate. This filtrate is then concentrated, sterilized, and spray-dried to obtain the Antarctic krill peptides. The amount of protease added is 4% of the mass of the minced meat.

[0026] The preparation method of perilla seed peptides includes: pulverizing perilla seed meal into fine perilla seed powder, adding water at a material-to-liquid ratio of 1 g: 10-15 mL, homogenizing, and simultaneously adding cellulase, pectinase, and lipase at 45-55 ℃ for 1-4 h, followed by enzyme inactivation at 95-100 ℃ for 20 min, cooling to 50-60 ℃, adding protease for 3-6 h, and inactivating the enzyme at 90-95 ℃ for 20 min. The solution is then filtered through a plate and frame filter to obtain the filtrate, which is concentrated, sterilized, and spray-dried to obtain perilla seed peptides. The mass ratio of cellulase, pectinase, and lipase is 3:2:0.5. The total mass of cellulase, pectinase, and lipase is 0.8-1.2% of the total mass of perilla seed meal. The amount of protease added is 0.5% of the total mass of the fine perilla seed powder.

[0027] The preparation method of flaxseed peptides includes: pulverizing flaxseed meal into fine flaxseed powder, adding water at a material-to-liquid ratio of 1 g: 10-15 mL, homogenizing, and simultaneously adding cellulase, pectinase, and lipase at 45-55 ℃ for 1-4 h, followed by enzyme inactivation at 95-100 ℃ for 20 min, cooling to 50-60 ℃, adding protease for 3-6 h, and inactivating the enzyme at 90-95 ℃ for 20 min. The mixture is then filtered through a plate and frame filter to obtain the filtrate, which is concentrated, sterilized, and spray-dried to obtain flaxseed peptides. The mass ratio of cellulase, pectinase, and lipase is 3:2:0.5. The total mass of cellulase, pectinase, and lipase is 0.8-1.2% of the total mass of the fine flaxseed powder. The amount of protease added is 0.5% of the total mass of the fine flaxseed powder.

[0028] The preparation method of Sacha indica peptides includes: crushing Sacha indica pulp into fine powder, adding water at a material-to-liquid ratio of 1 g: 10-15 mL, homogenizing, and simultaneously adding cellulase, pectinase, and lipase at 45-55 ℃ for 1-4 h, followed by enzyme inactivation at 95-100 ℃ for 20 min, cooling to 50-60 ℃, adding protease for 3-6 h, and inactivating the enzyme at 90-95 ℃ for 20 min. The mixture is then filtered through a plate and frame filter to obtain the filtrate, which is concentrated, sterilized, and spray-dried to obtain Sacha indica peptides. The mass ratio of cellulase, pectinase, and lipase is 3:2:0.5. The total mass of cellulase, pectinase, and lipase is 0.8-1.2% of the total mass of the fine Sacha indica powder. The amount of protease added is 0.5% of the total mass of the fine Sacha indica powder.

[0029] In this invention, the Schizophyllum commune has the accession number CGMCC No. 40388, the yeast has the accession number CCTCC NO:M2019324, and the Bacillus subtilis has the accession number CCTCC NO:M 20221156. Schizophyllum commune, yeast, and Bacillus subtilis have all been disclosed.

[0030] Example 1

[0031] The whitening and antioxidant composition base of Example 1 includes the following raw materials: 20 kg of Antarctic krill peptide, 5 kg of perilla seed peptide, 5 kg of flaxseed peptide, 8 kg of sacha indica peptide, 1 kg of Dendrobium officinale extract, and 15 kg of tea tree flower ferment.

[0032] The preparation method of Dendrobium officinale extract includes the following steps: 2 kg of Dendrobium officinale powder is added to 40 L of extraction solution, and ultrasonic extraction is performed at 50 W and 70 ℃ for 30 min to obtain a crude extract. The crude extract is centrifuged, and the supernatant is collected. The supernatant is heated at 60 ℃ to obtain an upper layer solution containing Dendrobium officinale extract and a lower layer solution containing the extract. The upper layer solution is freeze-dried to obtain the final extract. The extraction solution is an aqueous solution of a eutectic solvent, and the mass fraction of the eutectic solvent in the extraction solution is 60%. The eutectic solvent is composed of capric acid and tetracaine in a molar ratio of 1:1. The content of Dendrobium officinale polysaccharide in the Dendrobium officinale extract is 58%.

[0033] The preparation method of tea flower fermentation product includes the following steps: 20 kg of tea flower pollen is added to 400 L of sterile water, sterilized at 121 ℃ for 20 min, and cooled to 30 ℃ to obtain the primary fermentation substrate. The pH of the primary fermentation substrate is adjusted to 6, and then 1×10⁻⁶ viable bacteria are inoculated at a volume ratio of 7%. 8 The primary fermentation product was obtained by primary fermentation of *Schizophyllum commune* at CFU / g at 30 ℃ for 3 days at 150 rpm. The primary fermentation product was then sterilized at 121 ℃ for 20 min and cooled to 28 ℃ to obtain the secondary fermentation substrate. A mixed culture of yeast and *Bacillus subtilis* was inoculated into the secondary fermentation substrate at a volume ratio of 8% for secondary fermentation to obtain the secondary fermentation product. This secondary fermentation product was sterilized at 121 ℃ for 20 min, centrifuged, and freeze-dried to obtain the tea flower fermentation product. The viable yeast count was 5 × 10⁻⁶. 7 CFU / g, viable count of Bacillus subtilis is 5 × 10⁻⁶. 7 The yeast to Bacillus subtilis mass ratio was 1:4 (CFU / g). The secondary fermentation was carried out at 28 °C for 72 h at a rotation speed of 300 rpm.

[0034] The preparation method of the whitening and antioxidant composition base material in Example 1 includes the following steps: 10 kg of Antarctic krill peptide, 8 kg of Sacha indica peptide, 5 kg of flaxseed peptide, 5 kg of perilla seed peptide, 1 kg of Dendrobium officinale extract, 15 kg of tea flower ferment and the remaining 10 kg of Antarctic krill peptide are put into a mixer in sequence and mixed at 60 r / min for 30 min to obtain the product.

[0035] Example 2

[0036] The whitening and antioxidant composition base of Example 2 includes the following raw materials: 30 kg of Antarctic krill peptide, 10 kg of perilla seed peptide, 0.5 kg of flaxseed peptide, 1 kg of sacha indica peptide, 5 kg of Dendrobium officinale extract, and 25 kg of tea tree flower ferment.

[0037] The preparation method of Dendrobium officinale extract includes the following steps: 8 kg of Dendrobium officinale powder is added to 240 L of extraction solution, and ultrasonic extraction is performed at 100 W and 80 ℃ for 50 min to obtain a crude extract. The crude extract is centrifuged, and the supernatant is collected. The supernatant is heated at 70 ℃ to obtain an upper layer solution containing Dendrobium officinale extract and a lower layer solution containing the extract. The upper layer solution is freeze-dried to obtain the final extract. The extraction solution is an aqueous solution of a eutectic solvent, and the mass fraction of the eutectic solvent in the extraction solution is 68%. The eutectic solvent is composed of capric acid and tetracaine in a molar ratio of 1.5:1. The content of Dendrobium officinale polysaccharides in the Dendrobium officinale extract is 60%.

[0038] The preparation method of tea flower fermentation product includes the following steps: 40 kg of tea flower pollen is added to 1200 L of sterile water, sterilized at 121 ℃ for 20 min, and cooled to 25 ℃ to obtain the primary fermentation substrate. The pH of the primary fermentation substrate is adjusted to 6.5, and then 8% by volume is inoculated with a viable count of 1×10⁻⁶ cells / mL. 10 The primary fermentation product was obtained by primary fermentation of *Schizophyllum commune* at CFU / g at 25 ℃ for 5 days at 180 rpm. The primary fermentation product was then sterilized at 121 ℃ for 20 min and cooled to 30 ℃ to obtain the secondary fermentation substrate. A mixed culture of yeast and *Bacillus subtilis* was inoculated into the secondary fermentation substrate at a volume ratio of 5% for secondary fermentation to obtain the secondary fermentation product. This secondary fermentation product was sterilized at 121 ℃ for 20 min, centrifuged, and freeze-dried to obtain the tea flower fermentation product. The viable yeast count was 5 × 10⁻⁶. 8 CFU / g, viable count of Bacillus subtilis is 5 × 10⁻⁶. 8 The yeast to Bacillus subtilis mass ratio was 1:3 (CFU / g). The secondary fermentation was carried out at 30 °C for 24 h at a rotation speed of 100 rpm.

[0039] The preparation method of the whitening and antioxidant composition base material in Example 2 includes the following steps: 15 kg of Antarctic krill peptide, 1 kg of Sacha indica peptide, 0.5 kg of flaxseed peptide, 10 kg of perilla seed peptide, 5 kg of Dendrobium officinale extract, 25 kg of tea flower fermentation product and the remaining 15 kg of Antarctic krill peptide are put into a mixer in sequence and mixed at 50 r / min for 30 min to obtain the product.

[0040] Example 3

[0041] The whitening and antioxidant composition base of Example 3 includes the following raw materials: 28 kg of Antarctic krill peptide, 1 kg of perilla seed peptide, 10 kg of flaxseed peptide, 15 kg of sacha indica peptide, 0.05 kg of Dendrobium officinale extract, and 20 kg of tea tree flower ferment.

[0042] The preparation method of Dendrobium officinale extract includes the following steps: 1 kg of Dendrobium officinale powder is added to 30 L of extraction solution, and ultrasonic extraction is performed at 80 W and 75 ℃ for 60 min to obtain a crude extract. The crude extract is centrifuged, and the supernatant is collected. The supernatant is heated at 70 ℃ to obtain an upper layer solution containing Dendrobium officinale extract and a lower layer solution containing the extract. The upper layer solution is freeze-dried to obtain the final extract. The extraction solution is an aqueous solution of a eutectic solvent, and the mass fraction of the eutectic solvent in the extraction solution is 70%. The eutectic solvent is composed of capric acid and tetracaine in a molar ratio of 2:1. The content of Dendrobium officinale polysaccharides in the Dendrobium officinale extract is 62%.

[0043] The preparation method of tea flower fermentation product includes the following steps: 40 kg of tea flower pollen is added to 2000 L of sterile water, sterilized at 121 ℃ for 20 min, and cooled to 28 ℃ to obtain the primary fermentation substrate. The pH of the primary fermentation substrate is adjusted to 5.5, and then 1×10⁻⁶ viable bacteria are inoculated at a volume ratio of 5%. 9 The primary fermentation product was obtained by primary fermentation of *Schizophyllum commune* at CFU / g at 28 ℃ for 4 days at 160 rpm. The primary fermentation product was then sterilized at 121 ℃ for 20 min and cooled to 35 ℃ to obtain the secondary fermentation substrate. A mixed culture of yeast and *Bacillus subtilis* was inoculated into the secondary fermentation substrate at a volume ratio of 10% for secondary fermentation to obtain the secondary fermentation product. This secondary fermentation product was sterilized at 121 ℃ for 20 min, centrifuged, and freeze-dried to obtain the tea flower fermentation product. The viable yeast count was 5 × 10⁻⁶. 9 CFU / g, viable count of Bacillus subtilis is 5 × 10⁻⁶. 9 The yeast to Bacillus subtilis volume ratio was 1:2 (CFU / g). The secondary fermentation was carried out at 35 °C for 36 h at a rotation speed of 200 rpm.

[0044] The preparation method of the whitening and antioxidant composition base material in Example 3 includes the following steps: 14 kg of Antarctic krill peptide, 15 kg of Sacha indica peptide, 10 kg of flaxseed peptide, 1 kg of perilla seed peptide, 0.05 kg of Dendrobium officinale extract, 20 kg of tea tree flower fermentation product and the remaining 14 kg of Antarctic krill peptide are sequentially put into a mixer and mixed at 50 r / min for 30 min to obtain the product.

[0045] Comparative Example 1

[0046] The whitening and antioxidant composition base of Comparative Example 1 is largely the same as that of Example 1. The difference between the whitening and antioxidant composition base of Comparative Example 1 and Example 1 is that the whitening and antioxidant composition base of Comparative Example 1 removes the tea flower fermentation product and supplements the mass according to the mass ratio of each substance in Example 1.

[0047] Comparative Example 2

[0048] The whitening and antioxidant composition base of Comparative Example 2 is roughly the same as that of Example 1. The difference between the whitening and antioxidant composition base of Comparative Example 2 and Example 1 is that the whitening and antioxidant composition base of Comparative Example 1 removes Dendrobium officinale extract and supplements the mass according to the mass ratio of each substance in Example 1.

[0049] Comparative Example 3

[0050] The whitening and antioxidant composition base of Comparative Example 3 is largely the same as that of Example 1. The difference between the whitening and antioxidant composition base of Comparative Example 3 and Example 1 is that the whitening and antioxidant composition base of Comparative Example 3 removes the senna peptide and supplements the mass according to the mass ratio of each substance in Example 1.

[0051] Comparative Example 4

[0052] The whitening and antioxidant composition base of Comparative Example 4 is largely the same as that of Example 1. The difference between the whitening and antioxidant composition base of Comparative Example 4 and Example 1 is that the whitening and antioxidant composition base of Comparative Example 4 removes flaxseed peptides and supplements the mass according to the mass ratio of each substance in Example 1.

[0053] Comparative Example 5

[0054] The whitening and antioxidant composition base of Comparative Example 5 is largely the same as that of Example 1. The difference between the whitening and antioxidant composition base of Comparative Example 5 and Example 1 is that the whitening and antioxidant composition base of Comparative Example 5 removes perilla seed peptide and supplements the mass according to the mass ratio of each substance in Example 1.

[0055] Comparative Example 6

[0056] The whitening and antioxidant composition base of Comparative Example 6 is largely the same as that of Example 1. The difference between the whitening and antioxidant composition base of Comparative Example 6 and Example 1 is that the whitening and antioxidant composition base of Comparative Example 6 removes Antarctic krill peptides and supplements the mass according to the mass ratio of each substance in Example 1.

[0057] Application Example 1

[0058] 3 kg of panthenol, 0.5 kg of glycerol, 2.5 kg of sodium hyaluronate, 1.5 kg of xanthan gum, and 82.48 kg of water were stirred evenly at 45 °C and 300 rpm to obtain phase A. 3 kg of squalane, 3 kg of isononyl isononanoate, 1 kg of cetearyl alcohol, and 0.5 kg of polyglycerol-10 stearate were stirred evenly at 45 °C and 300 rpm to obtain phase B. Phase B was added to phase A, emulsified and homogenized, and cooled to 25 °C. Subsequently, 2.5 kg of the whitening and antioxidant composition base material from Example 1 and 0.02 kg of p-hydroxyacetophenone were added, and the mixture was stirred evenly to obtain a product with whitening and antioxidant properties.

[0059] The whitening and antioxidant composition bases of Examples 2-3 and Comparative Examples 1-6 were prepared according to the preparation method of Application Example 1 to produce whitening and antioxidant products of Application Examples 2-3 and Comparative Examples 1-6, respectively.

[0060] Example of effect

[0061] 1. Skin irritation test

[0062] Test Method: 270 volunteers were selected for testing, divided into 9 groups of 30 each, corresponding to the whitening and antioxidant products used in Examples 1-3 and Comparative Examples 1-6, respectively. Each participant used the inner sides of their left and right arms as controls. 0.3 g of the whitening and antioxidant products used in Examples 1-3 and Comparative Examples 1-6 were applied to the inner side of the left arm each time, while the right arm was left untreated as a control. This was done twice daily, and skin reactions were recorded after 24 hours and 72 hours.

[0063] The results showed that no skin redness or erythema or other allergic reactions were observed after 24 h and 72 h, indicating that the whitening and antioxidant composition base of the present invention is safe and has no adverse effects.

[0064] 2. DPPH removal rate

[0065] Test method: A 0.2 mmol / L DPPH solution was prepared by dissolving DPPH in anhydrous ethanol. Sample solutions of Examples 1 and Comparative Examples 1-6 with a concentration of 0.5 mg / mL were prepared. The DPPH solution and sample solutions were mixed in equal volumes and reacted at 25°C in the dark for 30 min. The absorbance at 517 nm was measured. The test was performed three times, and the average value was taken. The DPPH radical scavenging rate (%) = [1-(A1-A2) / A0]×100%. A0: No sample solution added, DPPH added; A1: Sample solution added, DPPH added; A2: Sample solution added, no DPPH added.

[0066] 3. Hydroxyl radical scavenging rate

[0067] Test method: Prepare sample solutions with a concentration of 0.5 mg / mL as in Examples 1 and Comparative Examples 1-6. Take 2 mL of the sample solution and 2 mL of 2.5 mmol / L FeSO4 solution, 2 mL of 2.5 mmol / L salicylic acid solution, and 2 mL of 2.5 mmol / L H2O2 solution. Stir in the dark at 37 ℃ and let stand for 30 min. Measure the absorbance at 510 nm as A1. Perform three tests and take the average value. Use Vc as a positive control. Hydroxyl radical scavenging rate (%) = [A0 - (A1 - A2)] / A0 × 100%, where A0: no sample added; A1: sample added; A2: H2O2 replaced with distilled water.

[0068] 4. Superoxide anion scavenging rate

[0069] To prepare sample solutions with a concentration of 0.5 mg / mL for Examples 1 and Comparative Examples 1-6, take 2 mL of the sample solution and add 2.25 mL of Tris. The reaction was terminated by adding 1 drop of 10 mol / L hydrochloric acid at 4 min after immersion in HCl buffer solution (pH 8.36) and 0.25 mL of 10 mol / L pyrogallol solution in a 25 ℃ water bath. The absorbance was measured at 420 nm using a UV spectrophotometer in three separate measurements, and the average value was taken. Superoxide anion scavenging rate (%) = [1] [(S1-S2) / (A1-A2)]×100%, where A1 is the spontaneous oxidation of pyrogallol in Tris-HCl solution to produce O2. - A1 is the absorbance value of the Tris-HCl solution; A2 is the absorbance value of the O2 removal solution; S1 is the absorbance value of the solution after O2 removal. - The absorbance value of the reaction; S2 is the absorbance value of the sample solution.

[0070] 5. Tyrosinase activity inhibition rate

[0071] Prepare 100 U / mL tyrosinase solution, 0.015 mol / L L-tyrosine solution, and 0.5 mg / mL sample solutions from Examples 1 and Comparative Examples 1-6 using 100 mM phosphate buffer solution at pH 6.8. Then, add 1.0 mL of sample solution, 0.5 mL of tyrosinase solution, and 2.5 mL of 100 mM phosphate buffer solution at pH 6.8 to a test tube and mix thoroughly. Heat in a 37 ℃ water bath for 10 min, then add 1.0 mL of L-tyrosine solution and continue heating in a 37 ℃ water bath for another 10 min. Measure the absorbance at a wavelength of 475 nm. Test 3 groups and take the average value. Tyrosinase activity inhibition rate (%) = [1 [(S1-S2) / (A1-A2)]×100%. Where, A1: absorbance without sample solution and with tyrosinase solution; A2: absorbance without sample solution and without tyrosinase solution; S1: absorbance with sample solution and with tyrosinase solution; S2: absorbance with sample solution and without tyrosinase solution.

[0072] 6. Whitening Test

[0073] Twenty-five participants, aged 25-40 years, were selected for the study. An ultraviolet-induced human skin melanization model was used to evaluate the skin whitening efficacy. A xenon arc lamp sunlight simulator (290-400 nm) with continuous spectral radiation capable of generating UVA+UVB wavelengths was employed. Seven skin areas (of the same size and region) on the back were marked for irradiation. After the irradiation experiment, the products from Application Example 1 and Control Application Examples 1-6 were applied to the test areas for 28 consecutive days. A skin color analyzer was used to detect and record the percentage decrease in melanin on days 7, 14, 21, and 28. The test results are shown in Table 2.

[0074] Table 1. Comparison of the effects of the base materials in Example 1 and Comparative Examples 1-6

[0075]

[0076] Table 2. Percentage decrease in melanin

[0077]

[0078] Table 1 shows that when tea flower fermentation is combined with other substances, the tyrosinase activity inhibition rate of the composition is significantly higher than that of tea flower fermentation alone, indicating that the substances in this invention have a compounding effect. Tables 1-2 show that the substances in this invention have a good synergistic effect; when all substances are present, the whitening and antioxidant composition base of this invention exhibits the strongest antioxidant, whitening, and melanin-reducing properties.

[0079] Experimental Example 1

[0080] The effects of different fermentation methods on tea flower fermentation products

[0081] Table 3. Effects of different fermentation methods on tea flower fermentation products

[0082]

[0083] As can be seen from groups 1-4, compared to simultaneous fermentation and fermentation using yeast + Bacillus subtilis followed by fermentation using Schizophyllum commune, fermentation using Schizophyllum commune first resulted in a higher tyrosinase activity inhibition rate. Therefore, this invention uses Schizophyllum commune for fermentation first. As can be seen from groups 3-5, fermentation using Schizophyllum commune alone first resulted in the highest tyrosinase activity inhibition rate. As can be seen from groups 5-7, compared to three-step fermentation, fermentation using Schizophyllum commune first, followed by co-fermentation with yeast and Bacillus subtilis, yielded the best results. Therefore, this invention uses the fermentation method of group 5.

[0084] Experimental Example 2

[0085] Effects of fermentation conditions on tea flower fermentation products

[0086] 1. Schizophyllum commune inoculation amount: The inoculation amount of the mixed yeast and Bacillus subtilis was controlled to be 2% of the volume of the secondary fermentation substrate, and the mass ratio of yeast to Bacillus subtilis was 1:1. The effect of Schizophyllum commune inoculation amount on tea flower fermentation product was observed according to 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10% (v / v). The results are shown in Table 4.

[0087] Table 4. Effects of Schizophyllum commune inoculation amount on tea flower fermentation products

[0088]

[0089] As shown in Table 4, with the increase of the amount of Schizophyllum commune inoculated, the inhibition of tyrosinase activity first increased, then gradually leveled off, and finally decreased. The highest inhibition rate of tyrosinase activity was observed when the amount of Schizophyllum commune inoculated was 7% (v / v) of the primary fermentation substrate.

[0090] 2. Mixed inoculation amount of yeast and Bacillus subtilis: The inoculation amount of Schizophyllum commune was controlled at 7% (v / v) of the primary fermentation substrate, and the mass ratio of yeast to Bacillus subtilis was 1:1. The effect of mixed inoculation amount on tea flower fermentation product was observed according to the mixed inoculation amount of 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10% (v / v). The results are shown in Table 5.

[0091] Table 5. Effect of mixed inoculum size on tea flower fermentation products

[0092]

[0093] As shown in Table 5, the inhibition of tyrosinase activity in tea flower fermentation material first increased with the increase of mixed bacteria inoculation, and then gradually slowed down. The inhibition rate of tyrosinase activity was highest when the mixed bacteria inoculation accounted for 8% of the total volume of the secondary fermentation substrate.

[0094] 3. Ratio of yeast to Bacillus subtilis: The amount of Schizophyllum commune inoculated was controlled at 7% (v / v) of the primary fermentation substrate, and the amount of mixed bacteria inoculated was 8% (v / v) of the secondary fermentation substrate. The yeast to Bacillus subtilis weight ratios were 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, and 1:4, respectively. The effect of the yeast to Bacillus subtilis weight ratio on the tea flower fermentation product was observed, and the results are shown in Table 6.

[0095] Table 6. Effect of the ratio of yeast to Bacillus subtilis on tea flower fermentation products.

[0096]

[0097] As shown in Table 6, when the weight ratio of yeast to Bacillus subtilis is 1:2-4, the tea flower fermentation product inhibits the activity of tyrosinase by more than 55%.

[0098] Other embodiments of this disclosure will readily occur to those skilled in the art upon consideration of the specification and practice of the disclosure herein. This disclosure is intended to cover any variations, uses, or adaptations of this disclosure that follow the general principles of this disclosure and include common knowledge or customary techniques in the art not disclosed herein. The specification and examples are to be considered exemplary only, and the true scope and spirit of this disclosure are indicated by the claims.

[0099] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. The scope of patent protection of the present invention is determined by the claims. Similarly, any equivalent structural changes made based on the content of the present invention's specification should also be included within the scope of protection of the present invention.

Claims

1. A whitening and antioxidant composition base, characterized in that, The following raw materials are included in the weight percentage calculation: Antarctic krill peptide 20-30 parts, perilla seed peptide 1-10 parts, flaxseed peptide 0.5-10 parts, sacha indica peptide 1-15 parts, Dendrobium officinale extract 0.05-5 parts, tea tree flower ferment 15-25 parts; The preparation method of the tea flower ferment includes the following steps: S1: Add sterile water to tea tree pollen, sterilize and cool to obtain a primary fermentation substrate; inoculate the primary fermentation substrate with Schizophyllum commune for primary fermentation to obtain a primary fermentation product; S2: After sterilizing and cooling the primary fermentation material, a secondary fermentation substrate is obtained. A mixture of yeast and Bacillus subtilis is introduced into the secondary fermentation substrate for secondary fermentation to obtain a secondary fermentation product. After sterilizing, centrifuging, and drying the secondary fermentation product, tea flower fermentation product is obtained.

2. The whitening and antioxidant composition base according to claim 1, characterized in that, For each gram of tea pollen in S1, add 20-50 mL of sterile water; adjust the pH of the primary fermentation substrate to 5.5-6.5 before inoculating with the *Schizophyllum commune* for fermentation; during the primary fermentation, the amount of *Schizophyllum commune* inoculated accounts for 5-8% of the volume of the primary fermentation substrate; the primary fermentation temperature is 25-30 ℃, the time is 3-5 days, and the rotation speed is 150-180 rpm; the viable count of the *Schizophyllum commune* is 1×10⁻⁶. 8 -10 10 CFU / g.

3. The whitening and antioxidant composition base according to claim 1, characterized in that, The amount of the mixed yeast and Bacillus subtilis strain introduced in S2 accounts for 5-10% of the volume of the secondary fermentation substrate, and the viable count of the yeast is 5 × 10⁻⁶. 7 -5×10 9 CFU / g, the viable count of the Bacillus subtilis is 5 × 10⁻⁶. 7 -5×10 9 CFU / g; the mass ratio of yeast to Bacillus subtilis is 1:2-4; the temperature of the secondary fermentation is 28-35 ℃, the time is 24-72 h, and the rotation speed is 100-300 rpm.

4. The whitening and antioxidant composition base according to claim 1, characterized in that, The sterilization temperature in S1 and S2 is 110-121 ℃, and the sterilization time is 15-20 min. After cooling to 25-30 ℃ in S1, the primary fermentation substrate is obtained, and after cooling to 28-35 ℃ in S2, the secondary fermentation substrate is obtained.

5. The whitening and antioxidant composition base according to claim 1, characterized in that, The preparation method of the Dendrobium officinale extract includes the following steps: adding Dendrobium officinale powder to the extraction solution, ultrasonically extracting to obtain a crude extract, centrifuging the crude extract and taking the supernatant, heating the supernatant and drying the upper layer solution to obtain the extract; the extraction solution is an aqueous solution of a eutectic solvent, and the mass fraction of the eutectic solvent in the extraction solution is 60-70%; the eutectic solvent is composed of capric acid and tetracaine in a molar ratio of 1-2:

1.

6. The whitening and antioxidant composition base according to claim 5, characterized in that, For every gram of Dendrobium officinale powder, add 20-30 mL of extraction solution; the ultrasonic extraction power is 50-100 W, the ultrasonic extraction temperature is 70-80 ℃, and the ultrasonic extraction time is 30-60 min; the supernatant is heated to a temperature of 60-70 ℃.

7. The whitening and antioxidant composition base according to claim 1, characterized in that, The Antarctic krill peptides contain >90% peptides with a molecular weight ≤5000 Da, ≥75% peptide content, and ≥80% protein content; the perilla seed peptides contain >80% peptides with a molecular weight ≤10000 Da, ≥40% peptide content, and ≥50% protein content; the flaxseed peptides contain >80% peptides with a molecular weight ≤10000 Da, ≥50% peptide content, and ≥60% protein content; and the sacha inchi peptides contain >85% peptides with a molecular weight ≤3000 Da, ≥65% peptide content, and ≥75% protein content.

8. A method for preparing the whitening and antioxidant composition base as described in claim 1, characterized in that, The process includes the following steps: mixing 50% by weight of Antarctic krill peptide, all by weight of Sacha indica peptide, all by weight of Perilla frutescens seed peptide, all by weight of flaxseed peptide, all by weight of Dendrobium officinale extract, all by weight of tea tree flower ferment, and the remaining 50% by weight of Antarctic krill peptide in sequence, and mixing at 50-60 r / min for 20-30 min to obtain the final product.

9. The use of the whitening and antioxidant composition base material as described in claim 1 in the preparation of skin care products with whitening and antioxidant properties.

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