Composition with tightening, anti-wrinkle and whitening effects as well as preparation method and application thereof

Abstract

The invention discloses a composition with firming, anti-wrinkle and whitening effects, the composition is prepared from the following components in parts by weight: 10-50 parts of a composite micelle carrier, 0.1-10 parts of dihydromyricetin and 0.1-50 parts of panthenol, and meanwhile, the invention also provides a preparation method and application of the composition. The solubility and bioavailability of the composition are improved through a process technology, the biological activity of the composition can be effectively protected, and the composition has good firming, anti-wrinkle and whitening effects and has a wide market prospect.

Description

Technical Field

[0001] The present invention discloses a composition, a preparation method and an application thereof, and specifically, a composition with firming, anti-wrinkle and whitening effects, a preparation method and an application thereof, belonging to the technical field of cosmetics. Background Art

[0002] Throughout the development of personal care products, consumer demand for a comprehensive range of firming, anti-wrinkle, and whitening products continues to rise. Traditional firming and anti-wrinkle products often rely on ingredients such as retinol and peptides. While retinol can promote collagen production, it is highly irritating and can easily cause adverse reactions such as redness and peeling, and has little effect on skin whitening. Peptide-based products also have limited anti-wrinkle effects and may lead to skin tolerance with long-term use, failing to meet consumer demand for long-term anti-wrinkle effects. Common whitening products often use hydroquinone and kojic acid as their main ingredients. Hydroquinone has potential cytotoxicity, posing a safety risk with long-term use, while kojic acid is unstable and easily ineffective due to light and temperature. Furthermore, both ingredients have little effect on firming and anti-wrinkle.

[0003] The application of plant extracts in this field is increasing, but the synergy between ingredients is poor. For example, while common grape seed extract has some antioxidant potential, its benefits in terms of skin firming, wrinkle reduction, and whitening are relatively limited, making it difficult to achieve a multi-benefit combination. Furthermore, the complex composition of plant extracts often leads to compatibility issues with other ingredients in product formulations, resulting in inefficient delivery of active ingredients and significantly reduced stability. Some plant-derived antioxidants, due to their unstable chemical structures, are susceptible to oxidative decomposition during storage, significantly reducing the product's effectiveness. Dihydromyricetin, a natural flavonoid, possesses potent antioxidant, anti-inflammatory, and multiple biological activities. However, in-depth research and application of effective firming, anti-wrinkle, and whitening compositions is extremely scarce. Currently, there is no mature technology to rationally combine dihydromyricetin with other active ingredients to fully realize its potential in improving skin elasticity, reducing wrinkles, and brightening the skin.

[0004] Therefore, it is very necessary to develop a composition that rationally combines copper chlorophyllin with plant oils and fats, improves their solubility and bioavailability through specific process technologies, effectively protects their biological activity, and has good firming, anti-wrinkle and whitening effects. Summary of the Invention

[0005] In response to the above-mentioned deficiencies, the purpose of the present invention is to provide a composition that rationally combines copper chlorophyllin with plant oils and fats, and improves their solubility and bioavailability through specific process technologies, thereby effectively protecting their biological activity and having good firming, anti-wrinkle and whitening effects. A preparation method and application of the composition are also provided.

[0006] To this end, the first technical solution provided by the present invention is as follows:

[0007] A composition with firming, anti-wrinkle and whitening effects consists of the following components in parts by weight: 10-50 parts of a composite micelle carrier, 0.1-10 parts of dihydromyricetin and 0.1-50 parts of panthenol.

[0008] Furthermore, the composite micelle carrier consists of active substances, glycerol (ethylhexanoate), silk fibroin, glycerol and water.

[0009] Furthermore, the active ingredient consists of copper chlorophyllin and sunflower seed oil.

[0010] Furthermore, the mass ratio of copper chlorophyllin to sunflower seed oil in the active ingredient is 3:17.

[0011] Furthermore, the mass ratio of the active substance, glycerol (ethylhexanoate), silk fibroin, glycerol and water in the composite micelle carrier is 1:2:2:(20-80):(20-80).

[0012] Furthermore, the mass ratio of dihydromyricetin to panthenol is 1:(1-5).

[0013] The second technical solution provided by the invention is: a method for preparing a composition with firming, anti-wrinkle and whitening effects, comprising the following steps:

[0014] A. Stir copper chlorophyllin, sunflower seed oil, and glyceryl (ethylhexanoate) at 45-50° C. using a stirrer at 500-800 rpm for 2-5 min until completely dissolved and clear, to obtain an oil phase;

[0015] B. Stir water, glycerol, and fibroin at 55-60° C. with a stirrer at 500-800 rpm for 5-10 minutes until completely dissolved to obtain an aqueous phase;

[0016] C. Cool the aqueous phase obtained in step B to 45-50° C., slowly add the oil phase obtained in step A to the oil phase obtained in step B while stirring, switch to homogenization and shearing at a speed of 8000-15000 rpm, and then perform high-pressure homogenization in an ultra-high pressure microfluidizer at a homogenization pressure of 20000-28000 PSI, circulate homogenization 2-5 times, and control the discharge temperature at 5-15° C. to obtain a composite micellar carrier;

[0017] D. In step C, the speed of adding the oil phase to the water phase is 5-10 mL / min, and the stirring speed is 500-800 rpm;

[0018] E. The obtained composite micelle carrier is stirred at room temperature at a speed of 500-800 rpm for 2-5 minutes, and is mixed evenly with dihydromyricetin and panthenol to obtain a composition with firming, anti-wrinkle and whitening effects.

[0019] The third technical solution provided by the invention is: an application of the above composition, wherein the composition is used in the preparation of cosmetics.

[0020] Furthermore, the cosmetic is a skin external preparation prepared by using the composition with anti-photodamage efficacy as an active component and adding conventional pharmaceutical or cosmetic excipients or auxiliary ingredients.

[0021] Furthermore, the composition is added in an amount of 0.01-20% in cosmetics.

[0022] Furthermore, the composition is added to cosmetics in an amount of 0.1-15%.

[0023] Furthermore, it is characterized in that the cosmetics are one or more of lotion, essence, cream, mask, and gel.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. This invention utilizes ultra-high pressure microfluidization technology to create a composite micellar carrier by combining copper chlorophyllin, sunflower seed oil, glyceryl (ethylhexanoate), silk fibroin, dihydromyricetin, and glycerol. This technology addresses the difficulty in mixing oil-soluble copper chlorophyllin with dihydromyricetin, panthenol, and water. It not only stabilizes the ingredients but also facilitates their efficient combination, achieving the excellent performance of the composite.

[0026] 2. The composite micellar carrier of the present invention is compounded with panthenol in a specific ratio, and the synergistic effect is significant. Dihydromyricetin and copper chlorophyllin are innovatively combined. Dihydromyricetin, with its powerful antioxidant and anti-inflammatory capabilities, can effectively inhibit tyrosinase activity, reduce melanin production, and achieve whitening effects. At the same time, its antioxidant properties can scavenge free radicals, delay skin aging, and help anti-wrinkle. The unique structure of copper chlorophyllin enhances the barrier function of the skin and synergizes with dihydromyricetin to tighten the skin. At the same time, dihydromyricetin, copper chlorophyllin, silk fibroin, and panthenol cooperate with each other. Silk fibroin is rich in various amino acids and can promote collagen synthesis and enhance skin elasticity. Panthenol penetrates into the bottom layer of the skin, replenishes moisture, and makes the skin hydrated and plump. The four work together to keep the skin hydrated and smooth while achieving firming, anti-wrinkle and whitening. The overall formula is mild, with good biocompatibility, greatly reducing the risk of allergies, and providing all-round care for the skin.

[0027] 3. This invention precisely controls the discharge temperature of the composite micelle carrier and ultrahigh-pressure microfluidizer technology to below 15°C, effectively avoiding the damage to the activity of copper chlorophyllin caused by high temperatures. Furthermore, this encapsulation process significantly enhances the stability of copper chlorophyllin, vegetable oils, and silk fibroin, ensuring their long-lasting activity during mixing and guaranteeing product quality and efficacy.

[0028] 4. The present invention abandons organic reagents such as ethanol and chloroform that are commonly used in traditional encapsulation technologies, which not only avoids harm to the human body and pollution to the environment, but also conforms to the concept of sustainable development. In addition, because the process is suitable for continuous scale-up production, it can significantly improve the production efficiency of traditional nanocarriers, while being environmentally friendly, helping the company achieve the goal of reducing costs and increasing efficiency. DETAILED DESCRIPTION

[0029] The following further describes the claims of the present invention in detail in conjunction with specific implementation methods, but does not constitute any limitation to the present invention. Any limited modifications made by anyone within the scope of protection of the claims of the present invention are still within the scope of protection of the claims of the present invention.

[0030] Example 1

[0031] A firming, anti-wrinkle and whitening composition containing dihydromyricetin is prepared from the following raw materials in percentage by weight: 0.3g copper chlorophyllin, 1.7g sunflower seed oil, 4g glyceryl (ethylhexanoate), 4g silk fibroin, 4g dihydromyricetin, 50g glycerol, 31g water and 5g panthenol.

[0032] The composition of this embodiment is prepared by the following preparation method:

[0033] Step 1: stirring copper chlorophyllin, sunflower seed oil, and glyceryl (ethylhexanoate) at 45° C. and 800 rpm for 5 minutes until completely dissolved and clear, to obtain an oil phase;

[0034] Step 2: Stir glycerol, dihydromyricetin, and fibroin at 55° C. and 800 rpm for 10 min until completely dissolved to obtain an alcohol phase;

[0035] Step 3: Cool the alcohol phase obtained in step 2 to 45°C, slowly add the oil phase in step 1 to the alcohol phase obtained in step 2 while stirring, the speed of adding the oil phase to the alcohol phase is 10 mL / min, the stirring speed is 800 rpm, switch to homogenization shearing at a speed of 15000 rpm, and then perform high-pressure homogenization through an ultra-high pressure microfluidizer with a homogenization pressure of 25000 PSI. The mixture is circulated and homogenized 5 times, and the discharge temperature is controlled at 10°C to obtain a composite micelle carrier.

[0036] Step 4: Stir the obtained composite micelle carrier at 800 rpm for 5 minutes at room temperature, and mix it with water and panthenol to obtain a firming, anti-wrinkle and whitening composition containing dihydromyricetin.

[0037] Example 2

[0038] A firming, anti-wrinkle and whitening composition containing dihydromyricetin is prepared from the following raw materials in percentage by weight: 0.2g of copper chlorophyllin, 1.13g of sunflower seed oil, 2.66g of glycerol (ethylhexanoate), 2.66g of fibroin, 2.66g of dihydromyricetin, 20g of glycerol, 65.69g of water and 5g of panthenol.

[0039] The composition of this embodiment is prepared by the following preparation method:

[0040] Step 1: stirring copper chlorophyllin, sunflower seed oil, and glyceryl (ethylhexanoate) at 45° C. and 800 rpm for 5 minutes until completely dissolved and clear, to obtain an oil phase;

[0041] Step 2: Stir glycerol, dihydromyricetin, and fibroin at 60° C. and 500 rpm for 8 minutes until completely dissolved to obtain an alcohol phase;

[0042] Step 3: Cool the alcohol phase obtained in step 2 to 50°C, slowly add the oil phase in step 1 to the alcohol phase obtained in step 2 while stirring, add the oil phase to the alcohol phase at a speed of 8 mL / min, stir at a speed of 500 rpm, switch to homogenization shearing at a speed of 8000 rpm, and then perform high-pressure homogenization by an ultra-high pressure microfluidizer at a homogenization pressure of 28000 PSI, cycle homogenization twice, and control the discharge temperature at 15°C to obtain a composite micelle carrier.

[0043] Step 4: Stir the obtained composite micelle carrier at 500 rpm for 3 minutes at room temperature, and mix it with water and panthenol to obtain a firming, anti-wrinkle and whitening composition containing dihydromyricetin.

[0044] Example 3

[0045] A dihydromyricetin-containing firming, anti-wrinkle and whitening composition is prepared from the following raw materials in percentage by weight: 0.1g copper chlorophyllin, 0.565g sunflower seed oil, 1.33g glyceryl (ethylhexanoate), 1.33g silk fibroin, 1.33g dihydromyricetin, 62.675g glycerol, 29.67g water and 3g panthenol.

[0046] The composition of this embodiment is prepared by the following preparation method:

[0047] Step 1: stirring copper chlorophyllin, sunflower seed oil, and glyceryl (ethylhexanoate) at 48° C. and 700 rpm for 2 min until completely dissolved and clear, to obtain an oil phase;

[0048] Step 2: Stir glycerol, dihydromyricetin, and fibroin at 58° C. and 700 rpm for 5 minutes until completely dissolved to obtain an alcohol phase;

[0049] Step 3: Cool the alcohol phase obtained in step 2 to 48°C, slowly add the oil phase in step 1 to the alcohol phase obtained in step 2 while stirring, add the oil phase to the alcohol phase at a speed of 5 mL / min, stir at a speed of 700 rpm, switch to homogenization shearing at a speed of 12000 rpm, and then perform high-pressure homogenization by an ultra-high pressure microfluidizer at a homogenization pressure of 20000 PSI, cycle homogenization 4 times, and control the discharge temperature at 5°C to obtain a composite micelle carrier.

[0050] Step 4: Stir the obtained composite micelle carrier at 700 rpm for 2 minutes at room temperature, and mix it with water and panthenol to obtain a firming, anti-wrinkle and whitening composition containing dihydromyricetin.

[0051] Comparative Example 1

[0052] In this comparative example, the ultrahigh pressure microfluidizer treatment in Example 1 was replaced with homogenous stirring using conventional technology, while other aspects remained unchanged.

[0053] Comparative Example 2

[0054] In this comparative example, the copper chlorophyllin in Example 2 was removed and the remaining amount was supplemented with water, while other conditions remained unchanged.

[0055] Comparative Example 3

[0056] In this comparative example, the dihydromyricetin in Example 2 was removed and the remaining amount was supplemented with water, while other contents remained unchanged.

[0057] Comparative Example 4

[0058] In this comparative example, the discharge temperature of the material treated by the ultrahigh pressure microfluidizer in Example 1 was not controlled, and other parameters were kept unchanged.

[0059] Comparative Example 5

[0060] In this comparative example, the silk fibroin in Example 2 was replaced with polysorbate 80, a relatively common emulsifier in cosmetics, while other ingredients remained unchanged.

[0061] Table 1. Composition of Examples and Comparative Examples

[0062]

[0063] In order to better illustrate the advantages of the present invention, the verification scheme provided by the present invention is given below:

[0064] Test Example 1: Accelerated stability performance test

[0065] This test is a one-month accelerated stability performance test of the compositions obtained in Examples 1-3 and Comparative Examples 1-5.

[0066] The composition prepared above was subjected to stability testing under normal temperature (room temperature), illumination (natural light), low temperature (-15°C), high temperature (45°C), high-low temperature cycling (45°C / -15°C for 24 hours each), and refrigeration (5°C). The data are recorded in Table 2.

[0067] Table 2. One-month accelerated stability performance test results of the composition

[0068]

[0069]

[0070] As can be seen from Table 2, there are no abnormalities in the accelerated stability of Examples 1-3 and Comparative Examples 2-3, while Comparative Examples 1, 4 and 5 all show unstable phenomena such as precipitation and fading stratification. The results show that with the help of ultra-high pressure micro-jet homogenization technology, silk fibroin is selected as the membrane material, and the composite micelle carrier prepared by strictly controlling the discharge temperature can effectively solve the stability problem faced when oil-soluble copper chlorophyllin is compounded with dihydromyricetin, panthenol and water. At the same time, this technical solution has a significant effect on delaying the fading problem of copper chlorophyllin. In addition, this experimental result further confirms that silk fibroin and copper chlorophyllin can only play their full role when organically combined with a specific ultra-high pressure micro-jet homogenization technology process, effectively solving the stability problem in the process of compounding copper chlorophyllin and dihydromyricetin, and providing a solid guarantee for the quality and performance of the product.

[0071] Test Example 2: Human skin patch test

[0072] This test involved the compositions obtained in Examples 1-3 and Comparative Examples 1-5, prepared as 10% aqueous solutions. Following the human skin patch test specified in the 2022 Cosmetic Safety Technical Specifications, skin reactions were observed at 30 minutes (after the indentation disappeared), 24 hours, and 48 hours, and the results were recorded. The data are summarized in Table 3.

[0073] Table 3. Human skin patch test results of the composition

[0074] serial number 30min 24h 48h Example 1 Level 0, 30 people Level 0, 30 people Level 0, 30 people Example 2 Level 0, 30 people Level 0, 30 people Level 0, 30 people Example 3 Level 0, 30 people Level 0, 30 people Level 0, 30 people Comparative Example 1 Level 0, 30 people Level 0, 30 people Level 0, 30 people Comparative Example 2 Level 0, 30 people Level 0, 30 people Level 0, 30 people Comparative Example 3 Level 0, 30 people Level 0, 30 people Level 0, 30 people Comparative Example 4 Level 0, 30 people Level 0, 30 people Level 0, 30 people Comparative Example 5 Level 0, 30 people Level 0, 30 people Level 0, 30 people

[0075] As shown in Table 3, the composition is non-irritating when added in an amount of up to 10%, and can be used safely within this range.

[0076] Test Example 3: Elastase Inhibition Test and Anti-wrinkle and Firming Efficacy Test

[0077] 3.1 Test Purpose and Principle

[0078] Anti-wrinkle and firming efficacy is primarily characterized by evaluating the test sample's inhibition rate against elastase. Elastase, primarily synthesized and secreted by fibroblasts, degrades elastin in the skin, contributing to skin aging. The experimental principle for elastase inhibition is a catalytic reaction between porcine pancreatic elastase and the enzyme substrate. Adding the active substance results in a change in absorbance, which reflects the inhibition rate of the elastase inhibitor.

[0079] This test refers to the laboratory method (HMC-WI-028 elastase inhibition rate) and compares the elastase inhibition rate test results of the test sample with that of the negative control. If the inhibition rate of the test sample is higher than that of the negative control and the difference is significant (P < 0.05), it can be considered that the test sample has certain anti-wrinkle and firming effects.

[0080] 3.2 Test indicators

[0081] Test indicator: elastase inhibition rate.

[0082] Judgment standard: If the elastase inhibition rate of the sample is higher than that of the negative control and there is a significant difference (p < 0.05), it can be considered that the test sample has certain anti-wrinkle and firming effects.

[0083] 3.3 Experimental materials and methods

[0084] Instruments: BSA224S analytical balance; Infinite200P20 microplate reader (Tecan)

[0085] Reagents: Elastase (porcine pancreas), BR; N-succinyl-L-alanyl-L-alanyl-L-alanine, 98%; Epigallocatechin gallate (EGCG), 98%

[0086] 3.4 Test method:

[0087] (1) Treatment of control and test samples

[0088] The compositions in Examples 1-3 and Comparative Examples 1-5 were diluted with pure water to a sample mass concentration of 2%; the positive control group (EGCG) was diluted with water to a positive control concentration of 0.1%; the negative control group was diluted with pure water.

[0089] (2) Test operation steps

[0090] Set up sample group, sample background group, solvent group and solvent background group. Each group needs to set up 3 parallels. Add different reagent solutions into the 96-well plate, shake gently, incubate at 25℃ for 15 minutes, place it in the microplate reader, and measure the absorbance at 410nm.

[0091] (3) Calculation formula

[0092] Elastase inhibition rate (%) = (1-(CD) / (AB))*100

[0093] Wherein: A is the absorbance of the reaction solution without sample; B is the absorbance of the reaction solution without sample and enzyme; C is the absorbance of the reaction solution with sample and enzyme; D is the absorbance of the reaction solution with sample and no enzyme.

[0094] (4) Data Analysis

[0095] Statistical analysis was performed using SPSS software. Comparisons of elastase inhibition rates between the test samples, positive controls, and negative controls were performed using independent sample t-tests. All statistical analyses were two-tailed, with a significance level of α = 0.05. A P > 0.05 indicated no significant difference between the two groups; a P < 0.05 indicated a significant difference between the two groups.

[0096] 3.5 Test results:

[0097] See Table 4.

[0098] Table 4. Elastase inhibition rate test results of the composition

[0099]

[0100]

[0101] As shown in Table 4, the anti-wrinkle and firming efficacy of the samples were tested in a laboratory method according to the laboratory method (HMC-WI-028 elastase inhibition rate); the test results show that the positive control elastase inhibition rate is greater than 50%, the reaction system is effective, and the elastase inhibition rate is significantly different from the negative control (p < 0.05), indicating that the samples have certain anti-wrinkle and firming efficacy. The elastase inhibition rates of the composition samples of Examples 1-3 are all above 60%, while the elastase inhibition rates of the composition samples of Comparative Examples 1-5 are all below 45%. The results show that by using ultra-high pressure microfluidization technology, selecting silk fibroin as the membrane material, and strictly controlling the discharge temperature, the composite micelle carrier prepared can fully release the anti-wrinkle and firming efficacy of the combination of copper chlorophyllin, dihydromyricetin and panthenol. This further confirms that only when silk fibroin and copper chlorophyllin are processed in a specific ratio and relying on a specific technical process can they exert a good anti-wrinkle and firming synergistic effect with panthenol.

[0102] Test Example 4: DPPH free radical scavenging rate and anti-wrinkle efficacy test

[0103] 4.1 Test Purpose and Principle

[0104] Excessive free radical production can lead to natural aging and photoaging of the skin, resulting in wrinkles. Therefore, the ability to scavenge free radicals is one of the important indicators for evaluating anti-aging and antioxidant cosmetics (raw materials).

[0105] This test refers to the laboratory method (HMC-WI-030 DPPH free radical scavenging rate) and compares the DPPH free radical scavenging rate test results of the test sample with that of the negative control. If the scavenging rate of the test sample is higher than that of the negative control and the difference is significant (P < 0.05), it can be considered that the test sample has a certain antioxidant effect.

[0106] 4.2 Test indicators

[0107] Test index: DPPH free radical scavenging rate

[0108] Judgment standard: If the DPPH free radical scavenging rate of the sample is higher than that of the negative control and there is a significant difference (p < 0.5), it can be considered that the test sample has a certain antioxidant effect.

[0109] 4.3 Experimental materials and methods

[0110] Instruments: BSA224S analytical balance; L6s UV spectrophotometer.

[0111] Reagent: DPPH (1,1-diphenyl 2-picrylphosphine), 98%

[0112] 4.4 Test methods

[0113] (1) Treatment of control and test samples

[0114] The compositions in Examples 1-3 and Comparative Examples 1-5 were diluted with pure water to a sample mass concentration of 2%; the positive control group (vitamin E, purity >96%) was diluted with 95% ethanol to a positive control concentration of 0.1%; the negative control group was diluted with pure water.

[0115] (2) Test operation steps

[0116] Set up three parallel tubes for each group: sample tube, sample background tube, DPPH tube, and solvent background tube. Add different reagent solutions to each of the four groups, shake gently, and let stand at room temperature for 5 minutes. Transfer each reaction solution into a 1 cm cuvette and measure the absorbance at 517 nm.

[0117] (3) Calculation formula

[0118] DPPH free radical scavenging rate (%) = (1-(T-T0) / (C-C0))*100%

[0119] In the formula: T-sample tube absorbance, that is, the absorbance of the solution after the sample reacts with DPPH; T0-sample background absorbance; C-DPPH tube absorbance value three times the average value, that is, the absorbance of the DPPH solution when no sample is added; C0-solvent background absorbance.

[0120] (4) Data Analysis

[0121] Statistical analysis was performed using SPSS software. Comparisons of DPPH radical scavenging rates between the test samples, positive controls, and negative controls were performed using independent sample t-tests. All statistical analyses were two-tailed, with a significance level of α = 0.05. A P > 0.05 indicated no significant difference between the two groups; a P < 0.05 indicated a significant difference between the two groups.

[0122] 4.5 Test results:

[0123] See Table 5.

[0124] Table 5. DPPH free radical scavenging rate test results of the composition

[0125] serial number DPPH free radical scavenging rate (%) P-value Example 1 93.18±4.62 ＜0.05 Example 2 88.62±3.54 ＜0.05 Example 3 84.07±3.27 ＜0.05 Comparative Example 1 46.18±2.51 ＜0.05 Comparative Example 2 32.16±1.87 ＜0.05 Comparative Example 3 30.75±2.06 ＜0.05 Comparative Example 4 43.27±2.12 ＜0.05 Comparative Example 5 38.24±2.08 ＜0.05 Negative control group 1.27±0.12 / Positive control group 90.16±4.12 ＜0.05

[0126] As shown in Table 5, the antioxidant efficacy of the samples was tested using a laboratory method (HMC-WI-030 DPPH radical scavenging rate). The test results showed that the positive control DPPH radical scavenging rate was >50%, indicating that the reaction system was effective. The average DPPH radical scavenging rate was significantly different from that of the negative control (p < 0.05), indicating that the product has certain antioxidant efficacy. The elastase inhibition rates of the compositions of Examples 1-3 were all above 80%, while the elastase inhibition rates of the compositions of Comparative Examples 1-5 were all below 50%. The results show that the composite micelle carrier prepared through ultrahigh-pressure microfluidization technology, the use of silk fibroin as the membrane material, and strict control of the discharge temperature can fully release the anti-wrinkle, firming, and antioxidant effects of the combination of copper chlorophyllin, dihydromyricetin, and panthenol. This further confirms that only when silk fibroin and copper chlorophyllin are processed in specific ratios and with specific technical processes can they exert a synergistic anti-wrinkle, firming, and antioxidant effect with panthenol.

[0127] Test Example 5: Tyrosinase Inhibition Rate and Whitening Efficacy Test

[0128] 5.1 Test Purpose and Principle

[0129] Tyrosinase is a key enzyme in the biosynthesis of melanin in the skin, acting on dopa to form dopaquinone, which then undergoes a series of reactions to form melanin. In a phosphate solution at pH 6.8, tyrosinase catalyzes the conversion of dopa to dopaquinone, with absorbance measured at 475nm on a spectrophotometer. Cosmetics that inhibit tyrosinase activity can reduce the conversion of dopa to dopaquinone, thereby lowering absorbance. Changes in absorbance can be used to assess the inhibitory effect of cosmetics on tyrosinase activity.

[0130] This test refers to the laboratory method (HMC-WI-032 tyrosinase inhibition rate), and the test samples and negative controls were tested for tyrosinase inhibition rate.

[0131] 5.2 Test indicators

[0132] Test index tyrosinase inhibition rate

[0133] Judgment standard: If the tyrosinase inhibition rate of the sample is higher than that of the negative control and there is a significant difference (P < 0.05), it can be considered that the test sample has a certain whitening effect.

[0134] 5.3 Test materials and methods

[0135] Instruments: BSA224S analytical balance; RT-6100 enzyme-labeled analyzer

[0136] Reagents: Polyphenol oxidase (mushroom), BR; L-dopa, BR

[0137] 5.4 Test methods

[0138] (1) Treatment of control and test samples

[0139] The compositions in Examples 1-3 and Comparative Examples 1-5 were diluted with pure water to a sample mass concentration of 2%; the positive control group (kojic acid, purity >96%) was diluted with pure water to a positive control concentration of 0.1%; the negative control group was diluted with pure water.

[0140] (2) Test operation steps

[0141] Set up sample tubes, sample background tubes, enzyme reaction tubes, and solvent background tubes. Set up three parallel tubes for each group. Add different reagent solutions to each of the four groups, shake gently, and let stand at room temperature for 5 minutes. Transfer each reaction solution into a 1 cm cuvette and measure the absorbance at 475 nm.

[0142] (3) Calculation formula

[0143] Tyrosinase inhibition rate (%) = (%) = (1-(T-T0) / (C-C0))*100%

[0144] Where: T-sample tube absorbance, i.e., the absorbance of the solution after the sample reacts with tyrosinase; T0-sample background absorbance; C-the average of three absorbance values ​​in the enzyme reaction tube, i.e., the absorbance of the reaction between tyrosinase and DOPA when no sample is added; C0-solvent background absorbance.

[0145] (4) Data Analysis

[0146] Statistical analysis software was SPSS, and the tyrosinase inhibition rates of the test samples, positive controls, and negative controls were compared using independent sample t-tests. All statistical analyses were two-tailed, with a significance level of α = 0.05. A P > 0.05 indicated no significant difference between the two groups, and a P < 0.05 indicated a significant difference between the two groups.

[0147] 5.5 Test results:

[0148] See Table 6.

[0149] Table 6. Tyrosinase inhibition rate test results of the composition

[0150] serial number Tyrosinase inhibition rate (%) P-value Example 1 82.17±3.16 ＜0.05 Example 2 80.23±2.83 ＜0.05 Example 3 78.27±2.66 ＜0.05 Comparative Example 1 26.18±1.72 ＜0.05 Comparative Example 2 19.64±1.14 ＜0.05 Comparative Example 3 17.59±1.35 ＜0.05 Comparative Example 4 23.15±1.64 ＜0.05 Comparative Example 5 21.35±1.27 ＜0.05 Negative control group 0.28±0.21 / Positive control group 88.47±3.69 ＜0.05

[0151] As can be seen from Table 6, the whitening efficacy of the samples was tested in a laboratory method according to the laboratory method (HMC-WI-032 tyrosine inhibition rate); the test results showed that the tyrosinase inhibition rate of the positive control group was >50%, the reaction system was effective, and the average tyrosinase inhibition rate was significantly different from that of the negative control (P < 0.05); indicating that the product has a certain whitening effect. The tyrosinase inhibition rates of the composition samples of Examples 1-3 were all above 75%, while the tyrosinase inhibition rates of the composition samples of Comparative Examples 1-5 were all below 30%. The results show that only by using ultra-high pressure microfluidization homogenization technology, selecting silk fibroin as the membrane material, and strictly controlling the discharge temperature, can the composite micelle carrier prepared fully release the whitening efficacy of the compound of copper chlorophyllin, dihydromyricetin and panthenol. This further confirms that only when silk fibroin and copper chlorophyllin are processed in a specific ratio and with the help of a specific technical process can they exert a good synergistic whitening effect with panthenol. The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A composition with firming, anti-wrinkle and whitening effects, characterized in that: The invention is composed of the following components in parts by weight: 10 to 50 parts of composite micelle carrier, 20 to 80 parts of water and 0.1 to 50 parts of panthenol.

2. The composition according to claim 1, characterized in that The composite micelle carrier consists of active substances, glycerol (ethylhexanoate), silk fibroin, glycerol and dihydromyricetin.

3. The composition according to claim 1, characterized in that The actives consist of copper chlorophyllin and sunflower seed oil.

4. The composition according to claim 1, characterized in that The mass ratio of copper chlorophyllin to sunflower seed oil in the active ingredient is 3:

17.

5. The composition according to claim 1, characterized in that The mass ratio of the active substance, glycerol (ethylhexanoate), silk fibroin, glycerol and water in the composite micelle carrier is 1:2:2:(20-80):(20-80).

6. The composition according to claim 1, characterized in that The mass ratio of the dihydromyricetin to panthenol is 1:(1-5).

7. A method for preparing a composition having firming, anti-wrinkle and whitening effects according to any one of claims 1 to 6, characterized in that: It consists of the following steps: A. Stir copper chlorophyllin, sunflower seed oil, and glyceryl (ethylhexanoate) at 45-50° C. using a stirrer at 500-800 rpm for 2-5 min until completely dissolved and clear, to obtain an oil phase; B. Stir water, glycerol, and fibroin at 55-60° C. with a stirrer at 500-800 rpm for 5-10 minutes until completely dissolved to obtain an aqueous phase; C. Cool the aqueous phase obtained in step B to 45-50° C., slowly add the oil phase obtained in step A to the oil phase obtained in step B while stirring, switch to homogenization and shearing at a speed of 8000-15000 rpm, and then perform high-pressure homogenization in an ultra-high pressure microfluidizer at a homogenization pressure of 20000-28000 PSI, circulate homogenization 2-5 times, and control the discharge temperature at 5-15° C. to obtain a composite micellar carrier; D. In step C, the speed of adding the oil phase to the water phase is 5-10 mL / min, and the stirring speed is 500-800 rpm; E. The obtained composite micelle carrier is stirred at room temperature at a speed of 500-800 rpm for 2-5 minutes, and is mixed evenly with dihydromyricetin and panthenol to obtain a composition with firming, anti-wrinkle and whitening effects.

8. Use of a composition having firming, anti-wrinkle and whitening effects according to any one of claims 1 to 6 in the preparation of cosmetics, characterized in that: The cosmetic is a skin external preparation prepared by taking the composition with firming, anti-wrinkle and whitening effects as an active component and adding conventional pharmaceutical or cosmetic excipients or auxiliary ingredients.

9. The use according to claim 8, characterized in that The composition is added in an amount of 0.01-20% in cosmetics.

10. The use according to claim 8, characterized in that The cosmetics are one or more of lotion, essence, cream, mask, and gel.

Images