An anti-aging composition, a method for preparing the same and use thereof in the cosmetic field

Abstract

The application belongs to the technical field of cosmetics, and provides an anti-aging composition, a preparation method thereof and application thereof in the cosmetic field.The anti-aging composition comprises rhodiola extract, radix paeoniae extract, ginkgo leaf extract and glutathione; the various plant extracts and the combination of active peptides and plant extracts have significant synergies, and are widely applied in the preparation of cosmetics with anti-aging effects; the composition can effectively improve skin aging problems from multiple aspects; in addition, the anti-aging composition is natural, mild and safe, the preparation method is simple, and the application is conducive to large-scale production.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, and relates to an anti-aging composition, a preparation method thereof and application thereof in the cosmetic field. BACKGROUND

[0002] The manifestations of skin aging on the skin mainly include dryness and roughness, pigmentation and dullness, relaxation and loss of elasticity, and wrinkles. Skin aging is a complex process of multiple factors, and can be divided into endogenous aging and exogenous aging according to different causes of aging. Among them, exogenous aging includes ultraviolet radiation, smoking, emotion and contact with toxic and harmful chemicals; endogenous aging includes DNA damage, cell damage and macromolecular damage, and DNA damage derives programmed aging theory and telomere theory.

[0003] Different anti-aging means are used for different manifestations of skin aging, for example, dryness and roughness can be solved by moisturizing and epidermal renewal; pigmentation and dullness can be solved by inhibiting melanin production; and wrinkles can be improved by stimulating continuous synthesis of collagen to fill the skin that has lost elasticity.

[0004] Chinese patent application CN116158988A discloses a skin care composition with whitening and anti-aging effects and application thereof, which comprises the following functional components: ergothioneine, VC ethyl ether and glutathione. The application first discovers that ergothioneine, VC ethyl ether, nicotinamide and glutathione have a significant synergistic effect on inhibiting tyrosinase activity. The components are scientifically matched to achieve the whitening and anti-aging effects of the product through multiple pathways. The raw materials of the composition can synergistically inhibit tyrosinase activity and scavenge free radicals. VC ethyl ether and glutathione have the effect of inhibiting the photodegradation of ergothioneine, which can effectively achieve the effects of whitening and anti-aging.

[0005] Chinese patent application CN116172901A discloses an anti-aging composition and a preparation method and application thereof. The anti-aging composition comprises the following components: ganoderma lucidum polysaccharide, astragalus polysaccharide, beta-nicotinamide mononucleotide, recombinant human lysozyme, carnosine and glutathione. The anti-aging composition effectively delays skin aging by enhancing cell proliferation capacity, inhibiting the expression of skin cell senescence associated secretory phenotype (SASP) factors, and enhancing cell antioxidant capacity.

[0006] Chinese invention patent application CN116265006A discloses a ginseng spot-removing anti-wrinkle essence, which comprises the following components: ginseng extract, superoxide dismutase, tocopherol, royal jelly, aloe extract, ganoderma extract, rhodiola kirilowii root extract, astragalus extract, honeysuckle flower extract and alcohol; the essence mainly comprises ginseng extract, and combines the effects of superoxide dismutase, tocopherol, royal jelly, aloe extract, ganoderma extract, rhodiola kirilowii root extract, astragalus extract and honeysuckle flower extract, so that the essence has the effects of spot removal, anti-wrinkle, alleviation of skin aging and moisturizing.

[0007] In the related anti-aging compositions disclosed in the prior art, the skin is usually only supplemented with collagen or melanin production is inhibited, the anti-aging path is relatively single, the effect is often poor, and the anti-aging effect cannot reach the expectation; meanwhile, there are problems such as relatively large number of traditional Chinese medicine extract components and high cost; therefore, it is urgent to develop a composition which can realize anti-aging effect from multiple paths such as free radical scavenging, inhibition of melanin production, promotion of collagen production and fine line lightening, so as to achieve synergistic anti-aging effect, prolong the action period of the composition, avoid rebound and comprehensively solve the skin aging problem. SUMMARY

[0008] In order to solve the problems of single anti-aging path, complex components and poor effect of the product in the prior art, the present application provides an anti-aging composition, a preparation method thereof and application thereof in the field of cosmetics, which has the effects of strong moisturizing, free radical scavenging, pigment deposition reduction, skin collagen production promotion, wrinkle lightening and firming, and can delay skin aging from multiple paths, comprehensively repair aging skin, and thus truly solve the skin aging phenomena such as wrinkles, relaxation, dryness, lack of water, dullness and the like.

[0009] One of the technical solutions of the present application is:

[0010] The present application provides an anti-aging composition, which comprises rhodiola extract, radix paeoniae alba extract, ginkgo leaf extract and glutathione.

[0011] The composition comprises, by weight fraction, 1-11 parts of rhodiola extract, 2-8 parts of radix paeoniae alba extract, 1-8 parts of ginkgo leaf extract and 9-12 parts of glutathione.

[0012] Preferably, the composition comprises, by weight fraction, 5-10 parts of rhodiola extract, 2-6 parts of radix paeoniae alba extract, 1-6 parts of ginkgo leaf extract and 9-11 parts of glutathione.

[0013] More preferably, the composition comprises, in parts by weight: 5 parts of Rhodiola extract, 4 parts of White Peony root extract, 1 part of Ginkgo leaf extract and 11 parts of glutathione.

[0014] The efficacy of each component of the anti-aging composition of the present application is as follows:

[0015] The Rhodiola extract contains a large amount of manganese ions, which can promote the activity of polysaccharide polymerase and inhibit the decomposition of collagen in the body. The Rhodioloside contained in the Rhodiola extract has strong antioxidant properties and can scavenge various free radicals, reduce oxidative stress damage, effectively absorb ultraviolet rays, and has a significant sunscreen effect. It can also inhibit the activity of tyrosinase, reduce the production of melanin, promote the proliferation of fibroblasts and the secretion of collagen, thereby effectively slowing down skin aging.

[0016] The White Peony root extract contains a variety of nutrients such as paeoniflorin, terpenes and flavonoids, which can inhibit UVA-induced photodamage, reduce cell apoptosis, reduce the production of ROS and malondialdehyde, and have the dual effects of whitening and delaying skin aging.

[0017] The flavonoids in the Ginkgo leaf extract are free radical scavengers that can protect dermal cells from excessive oxidative stress and reduce wrinkle formation by reducing the expression of MMP-1.

[0018] Glutathione contains imidazole rings and active sulfhydryl groups, which have strong physiological activity, can effectively scavenge free radicals, have antioxidant effects on the skin, and can also inhibit melanin production, thereby achieving the purpose of whitening and beautifying the skin.

[0019] The second technical solution of the present application is:

[0020] A preparation method of the above anti-aging composition is provided, which specifically comprises the following steps:

[0021] (1) Take the Rhodiola extract, White Peony root extract and Ginkgo leaf extract and add them into water, stir and dissolve to obtain a mixed solution;

[0022] (2) Adjust the pH of the mixed solution to 6.0-8.0;

[0023] (3) Add glutathione, stir for 1-2 hours, concentrate and dry to obtain the product.

[0024] The third technical solution of the present application is:

[0025] The above composition is provided for use in the preparation of cosmetics with anti-aging efficacy.

[0026] Further, the cosmetic is any one of toner, skin cream, emulsion, essence, facial mask, hand cream and eye cream.

[0027] The fourth technical solution of the present application is:

[0028] A skin cream is provided, which comprises the above-mentioned composition or the composition prepared by the above-mentioned preparation method and adjuvants.

[0029] The adjuvants comprise emulsifiers, thickening agents, humectants, oils, chelating agents, preservatives.

[0030] Further, the skin cream comprises, in percentage by mass: 0.01-10% of the anti-aging composition, 2-4% of the emulsifier, 0.3-0.5% of the thickening agent, 4-8% of the humectant, 11-20% of the oil, 0.03-0.05% of the chelating agent, 0.4-0.6% of the preservative, and the balance of water.

[0031] Preferably, the skin cream comprises, in percentage by mass: 0.01-2% of the anti-aging composition.

[0032] Further, the emulsifier is at least one of organic olive oil emulsified wax, self-emulsifying monoglyceride, methyl glucoside sesqui-stearate, PEG-20 methyl glucoside sesqui-stearate, and glycerol monostearate.

[0033] Further, the oil is at least one of caprylic capric triglyceride, shea butter, isohexadecane, isopropyl palmitate, dimethyl silicone oil, and cyclopentasiloxane.

[0034] Further, the thickening agent is at least one of carbomer U20, polyacrylate crosspolymer-6, and hydroxyethyl cellulose.

[0035] Further, the humectant is at least one of glycerol, propylene glycol, and sodium hyaluronate.

[0036] Further, the chelating agent is EDTA-2Na.

[0037] Further, the preservative is at least one of phenoxyethanol, hydroxybenzoic acid methyl ester, hydroxybenzoic acid propyl ester, and sodium benzoate.

[0038] The fifth technical solution of the present application is:

[0039] A preparation method of the above-mentioned skin cream is provided, which specifically comprises the following steps:

[0040] (1) accurately weigh the deionized water, thickening agent, humectant, and chelating agent, homogenize them, and keep them at 80-90℃ for 10-20min to obtain phase A;

[0041] (2) accurately weigh the oil and emulsifier, heat them to 80-90℃ and keep them for 10-20min to obtain phase B;

[0042] (3) Add B phase to A phase, homogenize at 8000-10000 rpm for 3-5 min;

[0043] (5) Stir and cool down, after the sample is cooled to 40-50℃, add preservatives and anti-aging composition;

[0044] (6) Continue to stir and cool down, and then cool down to room temperature.

[0045] Compared with the prior art, the present application has the following beneficial effects:

[0046] 1. In the anti-aging composition, the rhodiola extract can promote collagen synthesis and stabilize the collagen network structure of the skin, and has the effect of lightening and tightening; the ginkgo biloba extract has the effects of scavenging free radicals and antioxidation; the radix paeoniae alba extract has the effects of high-efficiency whitening and moisturizing; the glutathione has the effects of scavenging free radicals and high-strength antioxidation; the components synergize and complement each other, and through multiple ways such as moisturizing, scavenging free radicals, reducing pigmentation, promoting collagen production, lightening and anti-wrinkling, the purpose of resisting skin aging is achieved;

[0047] 2. The present application further improves the anti-aging effect of the composition by scientific matching of the dosages of the traditional Chinese medicine extracts and active peptides;

[0048] 3. The anti-aging skin cream contains the anti-aging composition, has multiple skin care effects, has a synergistic effect on skin aging, effectively resists skin aging and pigmentation, and makes the skin white, tender, elastic and lustrous;

[0049] 4. The natural anti-aging composition is mild and non-irritating, has a refreshing skin feel, is safe for long-term use, and the preparation method is simple and convenient, which is beneficial for large-scale production and application. BRIEF DESCRIPTION OF DRAWINGS

[0050] Figure 1 Results of Example 1 composition on HaCaT cell oxidative damage; compared with the blank group: # P<0.05; ## P<0.01; ### P<0.001; #### P<0.0001; compared with the model group: * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001;

[0051] Figure 2The results of the effect of the composition of Example 1 on tyrosinase activity; the model group compared with the blank group: #P<0.05; ##P<0.01; ###P<0.001; ####P<0.0001; the sample group compared with the model group: *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001;

[0052] Figure 3 The results of the effect of the composition of Example 1 on B16 melanoma cells; the model group compared with the blank group: #P<0.05; ##P<0.01; ###P<0.001; ####P<0.0001; the sample group compared with the model group: *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001;

[0053] Figure 4 The results of the effect of the composition of Example 1 on human fibroblasts; the model group compared with the blank group: #P<0.05; ##P<0.01; ###P<0.001; ####P<0.0001; the sample group compared with the model group: *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001;

[0054] Figure 5 The effect diagram of the eye wrinkle lightening after the skin cream of Application Example 1 is used by three subjects. DETAILED DESCRIPTION

[0055] The following non-limiting examples can make those skilled in the art more fully understand the present application, but do not limit the present application in any way. The following content is only an exemplary description of the scope claimed by the present application, and those skilled in the art can make various changes and modifications to the application disclosed, which should also belong to the scope claimed by the present application.

[0056] The raw materials of the present application are as follows:

[0057] The rhodiola extract (item number: YK2022062406), ginkgo leaf extract (item number: YK-SGSCF-YXYF), and white peony root extract (item number: YK-316) are all purchased from Xi'an Yungui Biotechnology Co., Ltd.

[0058] Glutathione (item number: LT5641) is purchased from Xi'an Huage Biotechnology Co., Ltd.

[0059] Example 1

[0060] An anti-aging composition comprises raw materials in the following weight proportions: Rhodiola extract 5 parts, white peony root extract 4 parts, ginkgo leaf extract 1 part, glutathione 11 parts.

[0061] Preparation method:

[0062] (1) Rhodiola extract, white peony root extract and ginkgo leaf extract are added to water, stirred and dissolved to obtain a mixture;

[0063] (2) The pH of the mixture is adjusted to 6.0-8.0;

[0064] (3) Glutathione is added, stirred and reacted for 1-2 h, concentrated and dried to obtain the product.

[0065] Example 2

[0066] An anti-aging composition comprises raw materials in the following weight proportions: Rhodiola extract 1 part, white peony root extract 6 parts, ginkgo leaf extract 8 parts, glutathione 10 parts.

[0067] The preparation method is the same as in Example 1.

[0068] Example 3

[0069] An anti-aging composition comprises raw materials in the following weight proportions: Rhodiola extract 9 parts, white peony root extract 2 parts, ginkgo leaf extract 7 parts, glutathione 12 parts.

[0070] The preparation method is the same as in Example 1.

[0071] Example 4

[0072] An anti-aging composition comprises raw materials in the following weight proportions: Rhodiola extract 10 parts, white peony root extract 8 parts, ginkgo leaf extract 2 parts, glutathione 9 parts.

[0073] The preparation method is the same as in Example 1.

[0074] Comparative Example 1

[0075] The difference from Example 1 is that a single component is used.

[0076] A plant extract comprises raw materials in the following weight proportions: Rhodiola extract 21 parts.

[0077] Preparation method: Rhodiola extract is used as an anti-aging composition.

[0078] Comparative Example 2

[0079] The difference from Example 1 is that a single component is used.

[0080] A plant extract comprises raw materials in the following weight proportions: white peony root extract 21 parts.

[0081] Preparation method: taking the extract of Radix Paeoniae Alba as the anti-aging composition.

[0082] Comparative Example 3

[0083] The difference from Example 1 is that a single component is used;

[0084] A plant extract comprising the following raw materials in parts by weight: Ginkgo biloba extract 21 parts.

[0085] Preparation method: taking the extract of Ginkgo biloba leaves as the anti-aging composition.

[0086] Comparative Example 4

[0087] The difference from Example 1 is that a single component is used;

[0088] A bioactive peptide comprising the following raw materials in parts by weight: glutathione 21 parts.

[0089] Preparation method: taking the bioactive peptide extract as the anti-aging composition.

[0090] Comparative Example 5

[0091] The difference from Example 1 is that it does not contain the extract of Rhodiola rosea;

[0092] An anti-aging composition comprising the following raw materials in parts by weight: extract of Radix Paeoniae Alba 8 parts, extract of Ginkgo biloba leaves 2 parts, glutathione 11 parts.

[0093] Preparation method same as Example 1.

[0094] Comparative Example 6

[0095] The difference from Example 1 is that it does not contain the extract of Radix Paeoniae Alba;

[0096] An anti-aging composition comprising the following raw materials in parts by weight: extract of Rhodiola rosea 8.33 parts, extract of Ginkgo biloba leaves 1.67 parts, glutathione 11 parts.

[0097] Preparation method same as Example 1.

[0098] Comparative Example 7

[0099] The difference from Example 1 is that it does not contain the extract of Ginkgo biloba leaves;

[0100] An anti-aging composition comprising the following raw materials in parts by weight: extract of Rhodiola rosea 5.56 parts, extract of Radix Paeoniae Alba 4.44 parts, glutathione 11 parts.

[0101] Preparation method same as Example 1.

[0102] Comparative Example 8

[0103] The difference from Example 1 is that it does not contain glutathione;

[0104] An anti-aging composition comprises the following raw materials in parts by weight: 5 parts of rhodiola extract, 4 parts of white peony root extract, and 1 part of ginkgo leaf extract.

[0105] Preparation method: directly mix the rhodiola extract, white peony root extract, and ginkgo leaf extract to obtain.

[0106] Comparative Example 9

[0107] The difference from Example 1 is that the rhodiola extract is replaced by ginseng extract with similar efficacy;

[0108] An anti-aging composition comprises the following raw materials in parts by weight: 5 parts of ginseng extract, 4 parts of white peony root extract, 1 part of ginkgo leaf extract, and 11 parts of glutathione.

[0109] The preparation method is the same as Example 1.

[0110] Comparative Example 10

[0111] The difference from Example 1 is that the white peony root extract is replaced by licorice extract with similar efficacy;

[0112] An anti-aging composition comprises the following raw materials in parts by weight: 5 parts of rhodiola extract, 4 parts of licorice extract, 1 part of ginkgo leaf extract, and 11 parts of glutathione.

[0113] The preparation method is the same as Example 1.

[0114] Comparative Example 11

[0115] The difference from Example 1 is that the ginkgo leaf extract is replaced by panax notoginseng extract with similar efficacy;

[0116] An anti-aging composition comprises the following raw materials in parts by weight: 5 parts of rhodiola extract, 4 parts of white peony root extract, 1 part of panax notoginseng extract, and 11 parts of glutathione.

[0117] The preparation method is the same as Example 1.

[0118] Comparative Example 12

[0119] The difference from Example 1 is that the glutathione is replaced by carnosine with similar efficacy;

[0120] An anti-aging composition comprises the following raw materials in parts by weight: 5 parts of rhodiola extract, 4 parts of white peony root extract, 1 part of ginkgo leaf extract, and 11 parts of carnosine.

[0121] The preparation method is the same as Example 1.

[0122] Comparative Example 13

[0123] The difference from Example 1 is that the amounts of the raw materials are different.

[0124] An anti-aging composition comprising the following raw materials in parts by weight: Rhodiola extract 12 parts, White Peony root extract 9 parts, Ginkgo leaf extract 1 part, and Glutathione 9 parts.

[0125] The preparation method is the same as that of Example 1.

[0126] Test Example 1: DPPH free radical scavenging rate test

[0127] The anti-aging composition prepared in each of the above examples and comparative examples is diluted with deionized water to a concentration of 0.25 mg / mL; 100 μL of the sample solution is added to a 96-well plate, and then 100 μL of 0.2 mmol / L DPPH anhydrous ethanol solution is added, and after uniform shaking, the mixture is placed in the dark at 25°C for 30 min, and then the absorbance A1 of the mixture is measured at a wavelength of 517 nm; 100 μL of anhydrous ethanol is used instead of the DPPH anhydrous ethanol solution to perform the same operation; 100 μL of anhydrous ethanol is used instead of the sample to perform the same operation; a 0.02 mg / mL vitamin C solution is used as a positive control, and 3 sets of parallel experiments are set up for each group to calculate the average value, and the DPPH free radical scavenging rate of the anti-aging repair composition in different groups is calculated according to the following formula:

[0128] DPPH scavenging rate = [1-(A1-A2) / A3] x 100%

[0129] In the formula:

[0130] A1 is the absorbance value of 100 μL of DPPH working solution + 100 μL of sample;

[0131] A2 is the absorbance value of 100 μL of anhydrous ethanol + 100 μL of sample;

[0132] A3 is the absorbance value of 100 μL of DPPH working solution + 100 μL of anhydrous ethanol.

[0133] The DPPH free radical scavenging rate results of the anti-aging compositions prepared in each of the examples and comparative examples are shown in Table 1.

[0134] Table 1: DPPH free radical scavenging rate test results

[0135]

[0136]

[0137] As can be seen from Table 1, the compositions obtained in Examples 1-4 all have good DPPH free radical scavenging capacity, and the highest scavenging rate reaches 97.08%, the DPPH free radical scavenging effect of the compositions obtained in Examples 1-4 is significantly better than that of any three components combined in Comparative Examples 1-4 using single component and Comparative Examples 5-8; the compositions obtained in Comparative Examples 9-12, although using components with similar efficacy to replace the corresponding components in the compositions of the application, have much lower DPPH free radical scavenging effect than the samples of Examples 1-4 of the application; the amount of Rhodiola and White Peony Root in the composition of Comparative Example 13 is slightly increased, and its DPPH free radical scavenging effect is lower than that of the composition of Example 1 of the application; it can be seen from this that the selected traditional Chinese medicine extracts and active peptides in the application have a synergistic effect within a certain dosage ratio range, and the four components are indispensable, and any component replaced by a component with the same efficacy cannot achieve the effect of the application; the composition of the application has strong free radical scavenging capacity and can effectively improve the skin aging problems such as skin dullness and yellowing caused by free radicals.

[0138] Test Example 2: Tyrosinase activity inhibition test

[0139] The anti-aging compositions prepared in each of the above examples and comparative examples were diluted with deionized water to a concentration of 0.25 mg / mL; 1 mL of sample solution and 1 mL of L-tyrosine solution were added to a 5 mL centrifuge tube, mixed and incubated at 37°C for 10 min; then 0.5 mL of tyrosinase solution was added and incubated for another 20 min; after incubation, 200 μL was taken to a 96-well plate, and the absorbance value was detected at 475 nm wavelength by an enzyme marker; 0.10 mg / mL arbutin was used as a positive control; the calculation formula of tyrosinase inhibition rate is:

[0140] Tyrosinase inhibition rate = [1-(T1-T2) / (T3-T4)] x 100%

[0141] In the formula:

[0142] T1 is the absorbance value of test sample + tyrosinase + L-tyrosine;

[0143] T2 is the absorbance value of test sample + L-tyrosine without adding tyrosinase;

[0144] T3 is the absorbance value of tyrosinase + L-tyrosine without adding sample;

[0145] T4 is the absorbance value of only L-tyrosine without adding sample and tyrosinase.

[0146] The test results of the tyrosinase activity inhibition rate of the anti-aging compositions prepared in each of the examples and comparative examples are shown in Table 2.

[0147] Table 2 Tyrosinase activity inhibition rate test results

[0148]

[0149]

[0150] As can be seen from Table 2, the compositions obtained in Examples 1-4 all have good tyrosinase activity inhibition ability, with the highest inhibition rate reaching 96.58%, and the tyrosinase activity inhibition effect of the compositions obtained in Examples 1-4 is significantly better than that of the single component used in Comparative Examples 1-4 and any three component combinations of Comparative Examples 5-8; the compositions obtained in Comparative Examples 9-12, although using components with similar efficacy to replace the corresponding components in the compositions of the application, have lower tyrosinase activity inhibition effect than the samples of Examples 1-4 of the application; the amount of Rhodiolae and Radix Paeoniae Alba in the composition of Comparative Example 13 is slightly increased, but its tyrosinase activity inhibition effect is lower than that of the composition of Example 1; it can be seen from this that the selected traditional Chinese medicine extracts and active peptides have a synergistic effect within a certain dosage ratio range, and the four components are indispensable, and any component replaced by a component with the same efficacy cannot achieve the effect of the application; the composition of the application has strong tyrosinase activity inhibition ability and can effectively improve the skin aging problems such as skin pigmentation caused by ultraviolet rays.

[0151] Test Example 3: Elastase activity inhibition test

[0152] The anti-aging compositions prepared in the above examples and comparative examples were diluted with deionized water to a concentration of 5 mg / mL; 1 mL of sample solution and 1 mL of elastase solution were added to a 5 mL centrifuge tube, mixed thoroughly, and then reacted at 37°C for 5 min; then 1 mL of substrate solution (N-methoxysuccinyl-ala-ala-pro-val-p-nitroanilide, MeOSuc-AAPV-pNA) was added, and the reaction was continued at 37°C for 60 min; after incubation, centrifugation was performed at 1000 rpm for 5 min, 200 μL of supernatant was taken to a 96-well plate, and the absorbance was detected at 405 nm wavelength by an enzyme marker, with 50 ng / mL of sivelestat sodium as a positive control. The calculation formula of elastase inhibition rate is as follows:

[0153] Elastase inhibition rate = [1-(Y1-Y2) / (Y3-Y4)] x 100%

[0154] In the formula:

[0155] Y1 is the absorbance value of the test sample + elastase solution + substrate solution;

[0156] Y2 is the absorbance value of the test sample + elastase solution + 10% DMSO;

[0157] Y3 is the absorbance value of the PBS buffer solution + elastase solution + substrate solution;

[0158] Y4 is the absorbance value of the PBS buffer solution + elastase solution + 10% DMSO.

[0159] The results of the elastase activity inhibition rate of the anti-aging composition prepared in each example are shown in Table 3;

[0160] Table 3 Test results of elastase activity inhibition rate

[0161] Group Elastase activity inhibition rate / % Positive control group 56.49 Example 1 66.95 Example 2 42.67 Example 3 52.67 Example 4 47.29 Comparative Example 1 23.57 Comparative Example 2 10.21 Comparative Example 3 12.35 Comparative Example 4 11.84 Comparative Example 5 32.67 Comparative Example 6 28.29 Comparative Example 7 25.64 Comparative Example 8 29.96 Comparative Example 9 36.57 Comparative Example 10 31.21 Comparative Example 11 34.35 Comparative Example 12 33.84 Comparative Example 13 37.69

[0162] As can be seen from Table 3, the compositions obtained in Examples 1-4 all have good elastase activity inhibition capacity, and the highest inhibition rate reaches 66.95%; the elastase activity inhibition effect of the compositions obtained in Examples 1-4 is significantly better than that of single component used in Comparative Examples 1-4 and any three component combinations of Comparative Examples 5-8; the compositions obtained in Comparative Examples 9-12, although using components with similar efficacy to replace a component in the composition of the present application, have lower elastase activity inhibition effect than the samples of Examples 1-4 of the present application; the use amount of Rhodiola and White Peony in the composition of Comparative Example 13 is slightly increased, but the elastase activity inhibition effect is lower than that of the sample of Example 1 of the present application; it can be seen that the selected traditional Chinese medicine extracts and active peptides in the present application have a synergistic effect within a certain dosage ratio range, and the four components are indispensable, and any component replaced by a component with the same efficacy cannot achieve the effect of the present application; the composition of the present application has strong elastase activity inhibition capacity, which can effectively improve the elasticity of elastic skin and delay skin relaxation and sagging and wrinkle formation.

[0163] In summary, the composition of the present application has high DPPH free radical scavenging rate, tyrosinase activity inhibition rate and elastase activity inhibition rate; the traditional Chinese medicine extracts Rhodiola extract, White Peony extract, Ginkgo leaf extract and active peptide glutathione in the composition can have a synergistic effect in inhibiting DPPH free radicals, tyrosinase and elastase activity.

[0164] Test Example 4: Cell activity test

[0165] (1) Determination of the effect of the composition of Example 1 on HaCaT cell oxidative damage:

[0166] HaCaT cells were plated in 96-well plates, divided into blank group, damage group and sample group, 3 parallel holes in each group. The blank group was given 100 μL of DMEM medium, the damage group was added with 100 μL of 250 μM H2O2; the sample group was first added with 100 μL of culture medium of different concentrations of the composition of Example 1 for pre-treatment for 24 h, and then 100 μL of 250 μM H2O2 was added for oxidative damage for 2 h; then the DMEM medium was replaced and incubated for 24 h; finally, the cell proliferation rate was detected by CCK-8, and the effect of different H2O2 concentrations on the cell proliferation rate was calculated; the calculation formula is as follows:

[0167] Cell proliferation rate = (A1-A2) / (A3-A2) x 100%

[0168] In the formula:

[0169] A1: absorbance value containing cells, CCK-8, sample and H2O2;

[0170] A2: absorbance value containing CCK-8, not containing cells, sample and H2O2;

[0171] A3: absorbance value containing cells, CCK-8, not containing sample and H2O2.

[0172] The determination results of the composition of Example 1 on HaCaT cell oxidative damage are shown in Table Figure 1 ; it can be seen from Figure 1 that compared with the blank group, the cell proliferation rate of the H2O2 model group was significantly reduced, with a statistical difference (P<0.0001), indicating that the modeling was successful; compared with the H2O2 model group, the cell proliferation rates of the cells treated with 200 μg / mL, 400 μg / mL and 800 μg / mL of the composition of Example 1 were significantly improved, which were (62.12±1.78) %, (74.73±2.18) % and (86.73±2.70) %, respectively, which were increased by 9.42% (P<0.001), 22.03% (P<0.0001) and 34.04% (P<0.0001) compared with the model group, respectively, with a statistical difference, indicating that the composition of Example 1 had good oxidative damage protection ability.

[0173] (2) Determination of the effect of the composition of Example 1 on B16 mouse melanoma cells:

[0174] The tyrosinase activity in B16 mouse melanoma cells was measured by L-DOPA substrate method. The B16 mouse melanoma cells were plated in 96-well plates according to the above method, and the B16 mouse melanoma cells were divided into a blank control group, a sample group and a positive control group, with 3 parallel holes in each group. The blank control group was added with 100 μL of RPMI1640 medium, the sample group was added with 100 μL of medium containing different concentrations of the composition sample of Example 1, and the positive control group was added with 100 μL of medium containing 0.1% VC. After incubation for 24 h, the cells were collected. The cell density was adjusted to 1.0 × 10 5 cells / 1 mL, and then the cells were placed in EP tubes. After centrifugation, the supernatant was discarded, 1 mL of 0.5% sodium deoxycholate solution was added, and vortex mixing was performed. Then, the cells were sequentially stored at 0°C for 20 min and placed at 37°C for 30 min. Then, the cell lysate was taken to a 96-well plate, 10 μL of 0.5% L-DOPA solution was added to each well, and incubation was performed at 37°C for 30 min. The absorbance value of each well was measured at 475 nm by an enzyme-labeled instrument. The tyrosinase activity in B16 melanoma cells was calculated according to the following formula:

[0175] Tyrosinase activity (%) = (OD sample / OD blank) × 100%

[0176] The relative content of melanin in B16 mouse melanoma cells was detected by NaOH method. The B16 mouse melanoma cells were plated in 96-well plates according to the above method, and the B16 mouse melanoma cells were divided into a blank control group, a sample group and a positive control group, with 3 parallel holes in each group. The blank control group was added with 100 μL of RPMI1640 medium, the sample group was added with 100 μL of medium containing different concentrations of the composition sample of Example 1, and the positive control group was added with 100 μL of medium containing 0.1% VC. Then, the cells were incubated in an incubator for 24 h, and then the supernatant was discarded, washed with PBS for 3 times, and 100 μL of 10% DMSO-containing 1 mol / L NaOH solution was added. The cell clumps were completely dissolved in a water bath at 80°C for 60 min. After cooling to room temperature, centrifugation was performed, and 150 μL of supernatant from each centrifuge tube was taken to a 96-well plate. The absorbance value was detected at 475 nm by an enzyme-labeled instrument, and the relative content of melanin was calculated according to the following formula:

[0177] Melanin relative content (%) = (OD sample / OD blank) × 100%

[0178] The determination results of the effect of the composition of Example 1 on B16 mouse melanoma cells are shown in Figure 2 , and the results of the relative content of melanin are shown in Figure 3 .

[0179] From Figure 2It can be seen that the tyrosinase inhibition rates were 28.18%, 45.19%, and 61.73% respectively compared with the blank group, with statistical difference (P<0.0001). With the increase of the concentration of the composition of Example 1, the activity of inhibiting tyrosinase of B16 melanoma cells was enhanced and showed a dose-dependent manner.

[0180] From Figure 3 It can be seen that the change trend of melanin content was consistent with the change trend of tyrosinase activity; compared with the blank group, it was reduced by 28.80%, 41.85%, and 71.32% respectively, with statistical difference (P<0.0001); with the increase of the concentration of the composition of Example 1, the synthesis of melanin of B16 melanoma cells was enhanced and showed a dose-dependent manner; therefore, the whitening mechanism of the composition of Example 1 was to reduce the content of melanin by inhibiting the activity of tyrosinase, thereby reducing the pigmentation of skin.

[0181] (3) Determination of the effect of the composition of Example 1 on human fibroblasts

[0182] Human fibroblasts were plated in a 96-well plate; divided into a blank group and a sample group, 3 parallel holes were set in each group, 100 μL of culture medium containing different concentrations of the composition of Example 1 was added to the sample group, 3 replicate wells were set for each concentration, the blank group was added with DMEM culture medium, and the positive control group was added with 100 μL of culture medium containing 0.1% VC, after incubation for 24 h, the cell supernatant was collected, and the contents of Collagen-Ⅰ, Elastase, MMP-1, and HA were detected by ELISA method, and the results are shown in Figure 4

[0183] From Figure 4 a、 Figure 4 It can be seen that compared with the blank group, the composition of Example 1 at 200 μg / mL, 400 μg / mL, and 800 μg / mL could significantly promote the synthesis of Collagen-Ⅰ and Elastase in a dose-dependent manner; from Figure 4 c, it can be seen that compared with the blank group, the composition of Example 1 at 200 μg / mL, 400 μg / mL, and 800 μg / mL significantly reduced the synthesis of MMP-1 in a dose-dependent manner; from Figure 4 d, it can be seen that compared with the blank group, the composition of Example 1 at 200 μg / mL, 400 μg / mL, and 800 μg / mL significantly increased the content of HA in a dose-dependent manner.

[0184] ​In summary, the composition of Example 1 can improve the proliferation activity of NHDF cells, increase the content of Collagen-I, Elastase and HA, inhibit the effect of MMP-1, maintain the skin water and elasticity state, and thus delay skin aging.

[0185] Application Example 1

[0186] The anti-aging composition obtained in Example 1 is used to prepare a skin cream, and the formula components of the skin cream are shown in Table 4.

[0187] Table 4 Formula of skin cream

[0188]

[0189]

[0190] The preparation method is as follows:

[0191] (1) accurately weigh each component of phase A, pre-disperse Cabo U20 with deionized water, homogenize uniformly, then add other components, and keep at 85℃ for 10 min;

[0192] (2) accurately weigh each component of phase B, heat to 85℃ and keep for 10 min;

[0193] (3) add phase B to phase A, and homogenize at 8000 rpm for 3 min;

[0194] (4) then, add the components of phase C;

[0195] (5) stir and cool, after the sample is cooled to 45℃, add each component of phase D;

[0196] (6) continue to stir and cool, and reduce to room temperature.

[0197] Comparative Application Example 1

[0198] The difference from Application Example 1 is only that the plant extraction composition of Example 1 is not contained, and other components and contents and the preparation method remain unchanged.

[0199] (1) Safety test:

[0200] The skin creams prepared by Application Example 1 and Comparative Application Example 1 are used as comparative objects, and the safety of the skin creams is verified by human skin patch test;

[0201] Test subjects: a total of 30 subjects, 15 males and 15 females; age distribution 18-55 years old.

[0202] Select an area of 49mm 2The samples prepared in Application Example 1 and Comparative Application Example 1 were respectively taken 0.025 g and put into the patch tester, and a blank control (without any sample) was set. The test samples and the blank control were all pasted on the flexor of the forearm of the testee, and the patch tester was uniformly pasted on the skin by pressing with the palm for 24 h. After the patch tester was removed, the results were recorded at 30 min (after the indentation disappeared), 24 h and 48 h according to the skin reaction grading standard (Table 5) in the Cosmetic Technical Specifications 2015 Edition.

[0203] Table 5 Skin reaction grading standard of skin occlusive patch test

[0204]

[0205]

[0206] The experimental results are shown in Table 6; from Table 6, it can be seen that the anti-wrinkle cosmetic prepared by the technical scheme of the present application has no adverse reaction on the human body, and the skin is negative in the patch test observed for 48 h, indicating that the composition prepared by the technical scheme of the present application is mild and non-irritating.

[0207] Table 6 Number of people with different grade reactions in skin occlusive patch test

[0208]

[0209] (2) Human skin moisture content test:

[0210] The skin care cream prepared in Application Example 1 and Comparative Application Example 1 was used as a comparative object, and the capacitance of the stratum corneum and the trans-epidermal water loss were measured before and after use by using the Torneometer probe and the Tewameter probe to measure the change in the moisturizing property of the skin care cream.

[0211] The change in the skin capacitance value reflects the moisture content (MMV) of the stratum corneum, and the greater the MMV value, the higher the moisture content of the stratum corneum; the trans-epidermal water loss (TEWL) represents a physical parameter of the normal and continuous diffusion of water through the stratum corneum, and is an important indicator reflecting the barrier function of the stratum corneum; skin aging can cause the stratum corneum to become thin, the skin barrier function to weaken, and the water loss to accelerate, and the TEWL value to increase.

[0212] Twenty-four healthy male and female subjects aged 18-65 years with a baseline forearm moisture content between 15-45 (Corneometer Units, CU) and no history of skin diseases or allergies were selected for the test. Before the test, subjects cleaned and dried the inner forearm and sat quietly for 30 minutes in an environment with a room temperature of (20±1)℃ and a relative humidity of (50±10)%. A 3×3cm sample was taken from each forearm. 2 A square experimental area was drawn, with the left arm serving as the test area for the skin cream in Application Example 1 and Control Application Example 1, using a concentration of 0.025 g / cm². 2 Apply the appropriate amount of sample to the test area and gently massage until it is completely absorbed. Use the symmetrical area of ​​the right arm as a blank control. Measure the stratum corneum moisture content (MMV) and transepidermal water loss (TEWL) of each test site at 0, 2, 4, and 8 hours after application. Each test was repeated 3 times, and the data were recorded. The test results are shown in Table 7.

[0213] Table 7. Results of Skin Stratum Corneum Moisture Content and Transepidermal Water Loss Tests

[0214]

[0215] As shown in Table 7, the sample with added Application Example 1 (containing 2% of the anti-aging composition of Example 1) had a much higher moisturizing effect than the blank group and the sample group without the anti-aging composition of Example 1. This indicates that the anti-aging plant extract composition provided by the present invention has a good moisturizing effect and can effectively improve the dryness and dehydration of the skin.

[0216] (3) Human skin whitening efficacy test:

[0217] Using the skin creams prepared in Application Example 1 and Comparative Application Example 1 as comparison objects, the melanin index and individual type angle before and after use were measured using the "CL400" probe and the "MX18" probe to evaluate the whitening effect of the skin creams.

[0218] The melanin index (MI) is a parameter that characterizes the melanin content in the skin by measuring the absorption of a specific wavelength of spectrum on the skin surface. The individual type angle (ITA°) is calculated from the three values ​​of the color system L*, a*, and b*. ITA° is a value that characterizes the skin color. The larger the ITA°, the brighter the skin, and vice versa.

[0219] Select 18-60 years old and the individual type angle ITA° value of the test site is 20-41°, which meets the test requirements. 30 qualified subjects are selected; determine the minimal erythema dose (MED) of each subject's test site through pre-test, establish a skin darkening model on the test site, use a solar simulator to irradiate at the same irradiation point with a dose of 0.75 times MED once a day for 4 consecutive days, and then enter the skin darkening period for 4 days after irradiation. Take one side as a blank control without applying any sample, and the other side as a sample group. After the success of the darkening model, apply 0.025g / cm 2 Apply the sample to the test area twice a day, morning and evening, until the end of the 8th week. After the success of the darkening model, test the skin melanin and colorimeter before the first application of the sample, which is recorded as the baseline value of the 0th week. After the test, apply the corresponding test material to each darkening test area once in the morning and once in the evening. Apply the test sample for 8 consecutive weeks, and test the skin melanin index MI and individual type angle ITA° after 2 weeks, 4 weeks and 8 weeks of application, and record them. The improvement rate of skin melanin index = (MI before use- MI after use) / MI before use, and the improvement rate of individual type angle = (ITA° after use- ITA° before use) / ITA° before use. The test results are shown in Table 8.

[0220] Table 8 Test results of skin melanin index and individual type angle

[0221]

[0222] As can be seen from Table 8, after using the skin cream of application example 1 (containing 2% of the anti-aging composition of example 1) for 8 weeks, the melanin index of the skin of the subjects decreased by more than 26.08%, and the individual type angle of the skin of the subjects increased by 16.40%; while the subjects using the comparative application example 1 product, the melanin index and individual type angle of the skin did not change significantly; further illustrating that the anti-aging composition provided by the present application has good effect on reducing pigmentation and brightening skin color.

[0223] (4) Human anti-wrinkle efficacy test:

[0224] The skin elasticity before and after use was measured using the MPA580 probe (Courage Khazaka, Germany) with the skin cream prepared in Application Example 1 and Comparative Application Example 1 as a comparative object. The skin was sucked into a specific probe under the negative pressure of the instrument, the depth of the sucked skin was determined by a non-contact optical testing system, and the elasticity of the skin was analyzed by MPA software. The ratio of the rebound amount of the skin without negative pressure to the maximum stretching amount of the skin with negative pressure was used for the evaluation of the elasticity of the skin. The closer the ratio is to 1, the better the elasticity of the skin. The anti-wrinkle effect of the skin cream was measured by taking facial images using a facial image analyzer VISIA-CR. The number of wrinkles around the eyes before and after applying the skin cream was analyzed. The fewer the number of wrinkles of the skin, the less or shallower the wrinkles.

[0225] Thirty healthy female subjects aged 35-50 years old were selected, who usually used skin care products and had visible wrinkles or fine lines around the eyes. The test site was cleaned with warm water and standard facial cleanser (laboratory formula) and the volunteers waited for 30 minutes in a specified room before collecting the baseline data of the cheek and eye sites. The volunteers used the product according to the product instructions, once a day in the morning and evening, and the use period was 8 weeks. The retest was conducted at the same time point after 4 weeks and 8 weeks of use. The improvement rate was calculated as follows: skin elasticity improvement rate = (after use - before use) / before use, and eye wrinkle improvement rate = (before use - after use) / before use. The results are shown in Table 9. The effect of eye wrinkles of three subjects using the skin cream of Application Example 1 (containing 2% of the anti-aging composition of Example 1) is shown in Figure 5

[0226] Table 9 Improvement rate of skin elasticity and number of eye wrinkles

[0227]

[0228]

[0229] As can be seen from Table 9, after using the skin cream of Application Example 1 (containing 2% of the anti-aging composition of Example 1) for 8 weeks, the elasticity of the skin of the subjects was improved by more than 39.39% compared with before use. The elasticity of the skin of the subjects using the product of Comparative Application Example 1 did not change significantly, which further indicates that the anti-aging composition provided by the application has a good effect on improving the elasticity of the skin.

[0230] As can be seen from Table 9 and Figure 5 ​It can be seen that the number of eye wrinkles is reduced by 33.01% after 8 weeks of using the skin care cream of application example 1 (containing 2% of the anti-aging composition of example 1), while the number of periorbital wrinkles of the subject using the comparative application example 1 product has no obvious change; in addition, it can be seen that the number of wrinkles of the test site is greatly improved over time, that is, the product provided by the present application can improve the skin wrinkles to a certain extent, and has excellent anti-wrinkle effect.

[0231] In summary, the anti-aging skin care cream of the present application contains an anti-aging composition, has multiple cosmetic skin care effects, has a synergistic effect on skin aging, effectively resists skin aging and color spots, and makes the skin white, tender, elastic and shiny; the natural composition of the anti-aging effect of the present application is mild and non-irritating, the skin feels fresh, and the long-term use is safe; in addition, the preparation method of the anti-aging cosmetic of the present application is simple and convenient, and is beneficial to large-scale production and application.

[0232] Finally, it should be noted that the above content is only used to illustrate the technical solutions of the present application, and is not a limitation on the protection scope of the present application. Simple modifications or equivalent replacements of the technical solutions of the present application made by those skilled in the art do not deviate from the essence and scope of the technical solutions of the present application.

Claims

1. An anti-aging composition characterized in that, The anti-aging composition comprises, by weight fraction, 1-11 parts of Rhodiola extract, 2-8 parts of White Peony root extract, 1-8 parts of Ginkgo biloba leaf extract, and 9-12 parts of glutathione.

2. Anti-aging composition according to claim 1, characterized in that, The anti-aging composition comprises, by weight fraction, 5-10 parts of Rhodiola extract, 2-6 parts of White Peony root extract, 1-6 parts of Ginkgo biloba leaf extract, and 9-11 parts of glutathione.

3. Anti-aging composition according to claim 2, characterized in that, The anti-aging composition comprises, by weight fraction, 5 parts of Rhodiola extract, 4 parts of White Peony root extract, 1 part of Ginkgo biloba leaf extract, and 11 parts of glutathione.

4. Process for the preparation of an anti-aging composition according to any one of claims 1-3, characterized in that, The method comprises the following steps: (1) Rhodiola extract, White Peony root extract, and Ginkgo biloba leaf extract are added to water, stirred and dissolved to obtain a mixture; (2) The pH of the mixture is adjusted to 6.0-8.0; (3) Glutathione is added, stirred and reacted for 1-2 hours, concentrated and dried to obtain the anti-aging composition.

5. Use of the anti-aging composition according to any one of claims 1 to 3 or the anti-aging composition prepared by the preparation method of claim 4 in the preparation of a cosmetic product having an anti-aging effect, characterized in that, The cosmetic is any one of toner, skin cream, emulsion, essence, and facial mask.

6. A skin cream characterized in that, According to mass percentage, the cosmetic comprises 2-4% emulsifier, 0.3-0.5% thickening agent, 4-8% humectant, 11-20% oil, 0.03-0.05% chelating agent, 0.4-0.6% preservative, 0.01-10% anti-aging composition of any one of claims 1-3 or prepared by the method of claim 4, and the balance of water.

7. The skin cream of claim 6, wherein, According to mass percentage, the anti-aging composition is 0.01-2%.

8. The skin cream according to claim 6 or 7, characterized in that, The emulsifier is at least one of organic olive oil emulsified wax, self-emulsifying monoglyceride, methyl gluceth-10-hydroxypropyl dimonium chloride, PEG-20 methyl gluceth-10-hydroxypropyl dimonium chloride, and glyceryl monostearate; The thickening agent is at least one of carbomer U20, polyacrylate crosspolymer-6, and hydroxyethyl cellulose; The humectant is at least one of glycerin, propylene glycol, and sodium hyaluronate; The oil is at least one of caprylic capric triglyceride, shea butter, isohexadecane, isopropyl palmitate, dimethicone, and cyclopentasiloxane; The chelating agent is EDTA-2Na; The preservative is at least one of phenoxyethanol, methylparaben, propylparaben, and sodium benzoate.

9. A method of making a skin cream as claimed in any one of claims 6 to 8, characterised in that, The method comprises the following steps: (1) Accurately weigh water, thickening agent, humectant, and chelating agent, homogenize, and keep at 80-90 ℃ for 10-20 min to obtain phase A; (2) Accurately weigh oil and emulsifier, heat to 80-90 ℃ and keep for 10-20 min to obtain phase B; (3) Add phase B to phase A and homogenize at 8000-10000 rpm for 3-5 min; (5) Stir and cool, and after the sample is cooled to 40-50 ℃, add preservative and anti-aging composition; (6) Continue to stir and cool, and after the temperature is reduced to room temperature, the cosmetic is obtained.

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