Rapid evaluation method for oil control efficacy of cosmetic raw materials

Abstract

The invention relates to the technical field of cosmetic detection, and discloses a rapid evaluation method for oil control efficacy of cosmetic raw materials, which comprises the following steps: 1) selecting a plurality of zebra fishes, and dividing the zebra fishes into a blank group and at least one sample group; incubating the blank group by using culture water, and incubating the sample group by using a sample solution containing a cosmetic raw material to be detected; 2) after the incubation is finished, respectively measuring the activity of 5 alpha-reductase, the content of bilayer methyl protein and the content of hyaluronic acid in each group of zebrafish bodies; 3, the oil control effect of the cosmetic raw material to be tested is scored according to the test result in the step 2. The oil control effect of the cosmetic raw material can be rapidly, comprehensively, objectively and accurately evaluated through scoring by taking the activity of 5 alpha-reductase, the content of bixymethyl protein and the content of hyaluronic acid in the zebra fish body as indexes based on the zebra fish model.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetics detection, and in particular to a method for quickly evaluating the oil-controlling efficacy of cosmetic raw materials. Background Art

[0002] With the changes in the pace of life and eating habits, excessive oil secretion has gradually become one of the skin problems that trouble people. Excessive oil secretion can cause a variety of skin problems such as enlarged pores, acne, acne, dermatitis, etc. Surveys and studies have shown that oily skin can have a significant negative impact on people's quality of life, and the market demand for oil-control cosmetics has therefore remained high. Therefore, the development of cosmetic raw materials with oil-control effects has always been a research hotspot in the cosmetics industry.

[0003] Existing non-human evaluation methods for oil control efficacy mainly include cell and animal models. Among them, the accuracy of the results of cell models is low due to the lack of organ or tissue interaction; animal model experiments are costly, have long test cycles, and are restricted by animal protection laws. As a small model organism, zebrafish has the characteristics of being highly similar to human genes, complete physiological system structure and function, fast development process and short test cycle, easy gene editing through in vitro fertilization, large egg production suitable for high-throughput screening, and transparent embryos that are easy to observe microscopic morphology. It is currently a popular evaluation model.

[0004] The group standard discloses an oil control efficacy evaluation method based on a zebrafish model, "Evaluation Method for Oil Control Efficacy of Cosmetics Based on Zebrafish Model" (T / QLMZ 7-2023). This method uses a zebrafish model to detect changes in zebrafish 5α-reductase (5α-reductase) related gene expression through fluorescent real-time quantitative PCR technology to evaluate the oil control efficacy of cosmetics. This method only targets the problem of sebaceous gland dysfunction caused by abnormal androgen hormones, resulting in excessive oil secretion; however, the present invention team has found through research that, in fact, skin oil secretion is not only related to sebaceous gland dysfunction, but also to impaired epidermal barrier function. If the epidermal barrier is damaged, the epidermis's ability to lock in water will deteriorate, and the sebaceous glands will increase oil secretion to protect the skin. Therefore, the evaluation indicators of the above method are relatively single and one-sided. At the same time, the operation steps of the fluorescent real-time quantitative PCR test are cumbersome, and the operating level of the test personnel is required to be high. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a rapid evaluation method for the oil control efficacy of cosmetic raw materials. The present invention is based on a zebrafish model, using 5α-reductase activity, loricrin content and hyaluronic acid content in zebrafish as indicators, and can quickly, comprehensively, objectively and accurately evaluate the oil control effect of cosmetic raw materials through scoring.

[0006] The specific technical scheme of the present invention is: a rapid evaluation method for the oil control efficacy of cosmetic raw materials, comprising the following steps: 1) selecting a number of zebrafish and dividing them into a blank group and at least one sample group; the blank group is incubated with aquaculture water, and the sample group is incubated with a sample solution containing the cosmetic raw material to be tested; 2) After incubation, the 5α-reductase activity, loricrin content, and hyaluronic acid content in zebrafish of each group were determined by ELISA; 3) Score the oil control efficacy of each tested cosmetic raw material based on the test results of 2).

[0007] The present invention team found in the previous study that abnormal male hormones and abnormal epidermal barrier function can cause vigorous skin oil secretion. Therefore, compared with the prior art that only uses 5α-reductase to characterize the function of sebaceous glands as an indicator, the present invention selects 5α-reductase, loricrin and hyaluronic acid as indicators to comprehensively evaluate the oil control effect of cosmetic raw materials, among which 5α-reductase is related to male hormone secretion, 5α-reductase can irreversibly convert testosterone into dihydrotestosterone, and dihydrotestosterone can induce excessive oil secretion of sebaceous glands. Loricrin and hyaluronic acid are related to epidermal barrier function. Loricrin is the main component of keratinocytes, accounting for about 80% of the total amount of keratinocytes, mainly present in the epidermal granular layer, and plays an important role in the barrier function of the epidermis; hyaluronic acid is the largest component of the extracellular matrix and has a very strong water binding ability.

[0008] It should also be emphasized that not all possible combinations of indicators that may be related to androgen abnormalities and epidermal barrier function abnormalities can accurately evaluate the oil-control efficacy of cosmetic raw materials. Different indicators have different degrees of correlation with different concentrations of cosmetic raw materials to be tested. Therefore, as an ideal indicator, it must be required to have a good correlation with the concentrations of most common cosmetic raw materials. Finally, the present invention has found through a large number of studies that only the combination of "5α-reductase activity + loricrin content + hyaluronic acid content" has a high correlation with most cosmetic raw materials within the maximum safe concentration range, and can objectively and accurately evaluate the oil-control efficacy.

[0009] Preferably, in 3), the oil control efficacy scoring criteria are as follows: Test value Scoring 5α-reductase activity ≤29.00 U / mL 4 >29.00 U / mL and ≤33.00 U / mL 2 >33.00 U / mL 0 Loricrin content ≥6.00 ng / mL 2 ≥5.50 ng / mL and <6.00 ng / mL 1 <5.50 ng / mL 0 Hyaluronic acid content ≥63.00 ng / mL 2 ≥58.00 ng / mL and <63.00 ng / mL 1 <58.00ng / mL 0 Among the three selected evaluation indicators, according to the different pathways of action, 5α-reductase activity is the most relevant indicator for male hormone secretion, so the full score of 5α-reductase activity is 4 points (the highest weight), and 0 points, 2 points and 4 points are given according to different test results; loricrin content and hyaluronic acid content are both indicators related to epidermal barrier function, so the full score of both is 2 points, and 0 points, 1 point and 2 points are given according to different test results. The sum of the three scores is recorded as the total score, 6 to 8 points are judged as good oil control effect, 3 to 5 points are judged as general oil control effect, and 2 points and below are judged as no oil control effect.

[0010] In the above table, the critical values ​​of the three indicators are determined based on the experience accumulated by the present invention team and a large amount of previous test data, and are highly scientific. Specifically, based on the repeated data of the blank group, the value with no significant difference from the blank group is used as the critical value for assigning a score of 0; through the test, multiple batches of cosmetic raw materials that are recognized to have significant oil control effects are measured, and based on the actual test data, 80%*actual test value is used as the critical point for assigning a full score.

[0011] Preferably, in 1), the incubation temperature is 25-30°C, fresh aquaculture water and sample solution are replaced every 20-30 hours, and the incubation time is 40-60 hours.

[0012] In order to more accurately reflect the 5α-reductase activity, loricrin content and hyaluronic acid content in zebrafish, it is necessary to further control the incubation time. The present invention has found through experiments that the 5α-reductase activity, loricrin content and hyaluronic acid content in zebrafish will show a significant change trend over time. Specifically: 5α-reductase activity showed a significant decrease on the 2nd day, which was significantly different from the 0th day (p<0.001), and the 5α-reductase activity continued to decrease on the 3rd and 4th days; while the loricrin content and hyaluronic acid content showed a significant increase trend on the 1st and 2nd days, and a slight decrease occurred on the 3rd and 4th days. According to the changes of the above 3 evaluation indicators at different times, taking into account the test cost, test efficiency and other factors, it is finally determined that the preferred time is about 2 days.

[0013] Preferably, in 1), the zebrafish is a 72 hpf wild-type AB strain zebrafish fry with normal development and consistent status.

[0014] The experiment chose zebrafish fry that developed to 72hpf because after developing to 72hpf, zebrafish fry can completely shed the egg membrane and become normal independent zebrafish fry individuals; at the same time, zebrafish within 5 days are not animals and can be regarded as animal alternative experiments.

[0015] Preferably, in 1), the method for obtaining the 72hpf wild-type AB strain zebrafish fry is: sexually mature wild-type AB strain zebrafish of different sexes are raised in separate tanks in a zebrafish circulation system, with a water temperature of 26±2°C; a pH value of 7-7.5; an electrical conductivity of 500-550μs / cm: a light / dark cycle of (12-16)h / (8-12)h, and a light intensity of 80-100W; the males and females are paired at a ratio of 1:(1.5-2.5) one day before mating, and natural mating and spawning are carried out, and the eggs are transferred into breeding water to cultivate 72hpf wild-type AB strain zebrafish fry.

[0016] Preferably, in 1), the number of zebrafish in each group is 20 to 30, and each group is provided with multiple parallel groups.

[0017] Preferably, in 1), the concentration of the cosmetic raw material to be tested in the sample solution is not higher than its maximum safe concentration, and the amount of each group of sample solutions is 3-5 mL.

[0018] Preferably, in 1), the standard for the maximum safe concentration of the cosmetic raw material to be tested in the sample solution is: the highest concentration at which more than 80% of the zebrafish fry after 3 days of cultivation show no mortality (no heartbeat) and other toxic effects (pericardial edema, trunk bending, no response to mechanical stimulation, unclear muscle texture, etc.) is the maximum safe concentration of the cosmetic raw material to be tested.

[0019] Preferably, in 1), the preparation method of the aquaculture water is: take 17-18g sodium chloride, 0.7-0.8g potassium chloride, 2.5-3.5g calcium chloride, 4.5-5g magnesium sulfate, dissolve them in water and make up to 1000mL, which is the aquaculture water reserve solution; take 16.67mL of the above aquaculture water reserve solution, and then dilute it to 1000mL with water; the amount of aquaculture water used in each group is 3-5mL.

[0020] Preferably, 2) specifically comprises: freezing the zebrafish, cutting into pieces, adding pre-cooled PBS, fully homogenizing, centrifuging the obtained homogenate, collecting the supernatant, and freezing and storing; and using ELISA to determine the 5α-reductase activity, loricrin content and hyaluronic acid content in each group of zebrafish.

[0021] Compared with the prior art, the beneficial effects of the present invention are: (1) Comprehensive evaluation indicators: The present invention conducts a comprehensive evaluation from two aspects: male hormones and epidermal barrier, which are significantly related to skin oil secretion. Three evaluation indicators, namely 5α-reductase, loricrin and hyaluronic acid, are selected, which is more comprehensive than the existing evaluation scheme. In addition, the scoring criteria of the present invention have the advantages of being scientific, reasonable and objective.

[0022] (2) The test efficiency is high. The method of the present invention only takes 5 days from collecting zebrafish eggs to the end of the test, which is shorter than other animal tests.

[0023] (3) Easy to operate: The sample processing method and index testing method of the present invention are relatively simple and do not require high technical skills from the experimenter.

[0024] (4) The present invention uses zebrafish as a model. Zebrafish lays a large number of eggs, is suitable for high-throughput screening, and has a low experimental cost. DETAILED DESCRIPTION

[0025] The present invention will be further described below in conjunction with the embodiments.

[0026] Overall embodiment A rapid evaluation method for the oil control efficacy of cosmetic raw materials, which specifically comprises the following steps: 1) Select a number of zebrafish and divide them into a blank group and at least one sample group; the blank group is incubated with aquaculture water, and the sample group is incubated with a sample solution containing the cosmetic raw material to be tested.

[0027] In some preferred implementation cases, the incubation temperature is 25-30° C., fresh aquaculture water and sample solution are replaced every 20-30 hours, and the incubation time is 40 to 60 hours.

[0028] In some preferred implementation cases, the zebrafish is a 72hpf wild-type AB strain zebrafish fry with normal development and consistent status, and the method for obtaining the zebrafish is as follows: sexually mature wild-type AB strain zebrafish are raised in separate tanks in a zebrafish circulation system with a water temperature of 26±2°C; a pH value of 7-7.5; an electrical conductivity of 500-550μs / cm: a light / dark cycle of (12-16)h / (8-12)h, and a light intensity of 80-100W; the males and females are paired at a ratio of 1:(1.5-2.5) the day before mating, and natural mating and spawning are carried out, and the eggs are transferred into the breeding water to cultivate 72hpf wild-type AB strain zebrafish fry.

[0029] In some preferred implementation cases, the number of zebrafish in each group is 20 to 30, and each group is provided with multiple parallel groups.

[0030] In some preferred implementation cases, the preparation method of the aquaculture water is: take 17-18g sodium chloride, 0.7-0.8g potassium chloride, 2.5-3.5g calcium chloride, 4.5-5g magnesium sulfate, dissolve in water and dilute to 1000mL, which is the aquaculture water reserve solution; take 16.67mL of the above aquaculture water reserve solution, and dilute to 1000mL with water; the amount of aquaculture water in each group is 3-5mL; the amount of each group is 3-5mL. The concentration of the cosmetic raw material to be tested in the sample solution is not higher than its maximum safe concentration, and the amount of each sample solution is 3-5mL. Among them, the standard of the maximum safe concentration of the cosmetic raw material to be tested in the sample solution is: the highest concentration of more than 80% of the zebrafish fry after 3 days of cultivation without any death (no heartbeat) and other toxic effects (pericardial edema, trunk bending, no response to mechanical stimulation, unclear muscle texture, etc.) is the maximum safe concentration of the cosmetic raw material to be tested.

[0031] 2) After incubation, ELISA was used to determine the 5α-reductase activity, loricrin content, and hyaluronic acid content in zebrafish of each group.

[0032] In some preferred implementation cases, 2) specifically includes: freezing the zebrafish, cutting into pieces, adding pre-cooled PBS, fully homogenizing, centrifuging the resulting homogenate, collecting the supernatant, and freezing and storing; using ELISA to determine the 5α-reductase activity, loricrin content, and hyaluronic acid content in each group of zebrafish.

[0033] 3) Score the oil control effect of each tested cosmetic raw material according to the test results of 2). The oil control effect scoring criteria are as follows: Test value Scoring 5α-reductase activity ≤29.00 U / mL 4 >29.00 U / mL and ≤33.00 U / mL 2 >33.00 U / mL 0 Loricrin content ≥6.00 ng / mL 2 ≥5.50 ng / mL and <6.00 ng / mL 1 <5.50 ng / mL 0 Hyaluronic acid content ≥63.00 ng / mL 2 ≥58.00 ng / mL and <63.00 ng / mL 1 <58.00ng / mL 0 The sum of the three scores is recorded as the total score. 6-8 points are considered to be good oil control effect, 3-5 points are considered to be fair oil control effect, and 2 points or less are considered to be no oil control effect. Specific embodiments The present invention will be further described in detail below in conjunction with specific examples. The following examples are not intended to limit the present invention, but are only intended to illustrate the present invention. The experimental methods used in the following examples are generally conventional, unless otherwise specified, and the materials, reagents, etc. used in the following examples are commercially available, unless otherwise specified.

[0035] Zebrafish 5α-reductase ELISA kit, zebrafish loricrin (LOR) ELISA kit, and zebrafish hyaluronic acid (HA) ELISA kit were purchased from Wuhan Aidikang Biotechnology Co., Ltd.

[0036] The fish used in the experiment were wild-type AB strain zebrafish purchased from Nanjing Yishulihua Biotechnology Co., Ltd.

[0037] The preparation method of aquaculture water is as follows: take 17.2g sodium chloride, 0.76g potassium chloride, 2.9g calcium chloride, and 4.9g magnesium sulfate, dissolve them in water and make up to 1000mL, which is the aquaculture water stock solution. Take 16.67mL of the above aquaculture water stock solution and dilute it to 1000mL with water.

[0038] Example 1: Acquisition of zebrafish embryos Sexually mature wild-type AB strain zebrafish were raised in separate tanks in a zebrafish circulation system. The water temperature was 26±2℃; the pH value was 7.2; the conductivity was 520μs / cm; the light / dark cycle was 14h / 10h, and the light intensity was 90W; the male and female were paired in a 1:2 ratio the day before mating, and natural mating and spawning were carried out. The eggs were transferred to the culture water.

[0039] Example 2: Selection of experimental fish The experiment chose zebrafish fry that developed to 72hpf because after developing to 72hpf, zebrafish fry can completely shed the egg membrane and become normal independent zebrafish fry individuals; at the same time, zebrafish within 5 days are not animals and can be regarded as animal alternative experiments.

[0040] Example 3: Determination of the maximum safe concentration of the test substance. Select zebrafish fry that have grown to 72 hpf and place them in a six-well plate. The six-well plate is divided into three groups: a blank group and a test sample group (3 to 5 appropriate concentrations are configured according to the characteristics of the test substance itself). Each concentration contains 30 zebrafish fry, and 10 mL of solution is added to each well. After 3 days, the hatching rate and mortality rate are observed. The maximum safe concentration of the test substance is the highest concentration at which more than 80% of the zebrafish fry do not show any death (no heartbeat) and other toxic effects (pericardial edema, trunk bending, no response to mechanical stimulation, unclear muscle texture, etc.) after 3 days.

[0041] Example 4: Selection of evaluation indicators Abnormal male hormones and abnormal epidermal barrier function can cause vigorous skin oil secretion. The present invention selected 8 related indicators for determination, tested tannic acid, salicylic acid and zinc pyrrolidone carboxylate, three raw materials recognized as having oil control effects, and analyzed the correlation between the concentration of each test sample and the 8 indicators, and selected the indicators with the top three correlation coefficients as the best evaluation indicators of the method. The results are shown in Tables 1, 2, 3, 4, 5 and 6.

[0042] Table 1: Tannic acid test results Table 2: Correlation analysis between tannic acid concentration and various test indicators Table 3: Salicylic acid test results Table 4: Correlation analysis between salicylic acid concentration and various test indicators Table 5: Test results of zinc pyrrolidone carboxylate Table 6: Correlation analysis between zinc pyrrolidone carboxylate concentration and various test indicators In order to find the most relevant index for evaluating the oil control effect, the inventor selected 8 indexes that may be related to abnormal male hormones and abnormal epidermal barrier function for testing, namely 5α-reductase activity, testosterone content, loricrin content, filamentous protein content, transglutaminase, aquaporin 3 content, hyaluronic acid content, and type I collagen content. And tannic acid, salicylic acid and zinc pyrrolidone carboxylate 3 kinds of raw materials recognized to have oil control effect were tested, and the correlation coefficient between the test index and the test sample concentration was analyzed. It can be seen from Tables 1 to 6 that the correlation coefficients between the 8 indexes and the test sample concentration are different, among which 5α-reductase activity, loricrin content and hyaluronic acid content have the highest correlation coefficient with the concentration of the test sample, and the absolute value of r exceeds 0.9, among which 5α-reductase activity is negatively correlated with the test sample concentration, and loricrin content and hyaluronic acid content are positively correlated with the concentration of the test sample. Therefore, it is determined to select 5α-reductase activity, loricrin content and hyaluronic acid content as indicators for evaluating the oil control effect.

[0043] Example 5: Determination of administration time The steps for obtaining zebrafish fry are the same as those in Example 1.

[0044] Healthy wild-type AB strain zebrafish larvae developed to 72 hpf were selected for experimental research and placed in a 6-well cell culture plate. 30 zebrafish were placed in each well and divided into a blank group and a positive control group (10 μg / mL tannic acid), with 3 parallels in each group. 5 mL of ordinary culture water was added to the blank group wells, and 5 mL of 10 μg / mL tannic acid solution was added to the positive control group wells; the cells were placed in a 28°C incubator for incubation, and fresh culture water and sample solution were replaced every 24 hours. The incubation time was 1, 2, 3, and 4 days, respectively. After the administration, the experimental zebrafish samples were processed according to the following method: the zebrafish were frozen at -20°C, weighed, cut into pieces for later use, and an appropriate amount of pre-cooled PBS was added at a weight-to-volume ratio of 1 g: 9 mL. The homogenate was fully homogenized in an ice bath using a glass homogenizer, and the homogenate was aspirated into a centrifuge tube, centrifuged at 5000×g for 5 minutes at 5°C, and the supernatant was collected and stored at -20°C. According to the instructions of the ELISA kit, the 5α-reductase activity, loricrin content and hyaluronic acid content in zebrafish were determined. The results are shown in Tables 7-9.

[0045] Table 7: 5α-reductase activity Table 8: Loricrin content Table 9: Hyaluronic acid content As can be seen from Tables 7 to 9, after zebrafish fry were treated with the positive control drug tannic acid (10 μg / mL), the 5α-reductase activity, loricrin content and hyaluronic acid content in the body showed a significant change trend. The 5α-reductase activity in zebrafish showed a significant decrease on the second day of administration, which was significantly different from that on the 0th day (p<0.001). The 5α-reductase activity continued to decrease on the third and fourth days of administration. The loricrin content and hyaluronic acid content in zebrafish showed a significant increase trend on the first and second days of administration, and a slight decrease on the third and fourth days of administration. According to the changes of the above three evaluation indicators at different administration times, considering the test cost, test efficiency and other factors, the administration time was selected as 2 days.

[0046] Example 6: Oil control efficacy grading evaluation method Among the three selected evaluation indicators, according to the different pathways of action, 5α-reductase activity is the most relevant indicator for male hormone secretion, so the full score of 5α-reductase activity is 4 points, and 0 points, 2 points and 4 points are given according to different test results; loricrin content and hyaluronic acid content are both indicators related to epidermal barrier function, so the full score of both is 2 points, and 0 points, 1 point and 2 points are given according to different test results. Specifically, according to Table 9, the three scores are added together as the total score, and the total score is 8 points. 6-8 points are judged as good oil control effect, 3-5 points are judged as general oil control effect, and 2 points or less are judged as no oil control effect.

[0047] Table 10: Oil control efficacy grading system In Table 10, the critical values ​​of the three indicators are determined based on the experience accumulated by the present invention team and a large amount of previous test data, and are highly scientific. Specifically, based on the repeated data of the blank group, the value with no significant difference from the blank group is used as the critical value for assigning a score of 0; through the test, multiple batches of cosmetic raw materials recognized to have significant oil control effects are measured, and based on the actual test data, 80%*actual test value is used as the critical point for assigning a full score.

[0048] Example 7: Evaluation of the oil control efficacy of niacinamide, tea polyphenols and vitamin C The steps for obtaining zebrafish fry are the same as those in Example 1.

[0049] The steps for determining the maximum safe concentration of the test substance are the same as in Example 3.

[0050] (1) Blank group: Healthy wild-type AB strain zebrafish fry developed to 72 hpf were selected for experimental research and placed in a 6-well cell culture plate, with 30 zebrafish fry placed in each well, and incubated with normal culture water.

[0051] (2) Nicotinamide sample group: Healthy wild-type AB strain zebrafish larvae that had developed to 72 hpf were selected for experimental research and placed in a 6-well cell culture plate, with 30 zebrafish placed in each well, and incubated with 0.5 mg / mL nicotinamide solution.

[0052] (3) Tea polyphenol sample group: Wild-type AB strain zebrafish larvae that developed to 72 hpf were selected for experimental research and placed in a 6-well cell culture plate with 30 zebrafish placed in each well. They were incubated with 1.0 mg / mL tea polyphenol solution.

[0053] (4) Vitamin C sample group: Healthy AB zebrafish larvae that had developed to 72 hpf were selected for experimental research and placed in a 6-well cell culture plate, with 30 zebrafish placed in each well, and incubated with 0.1 mg / mL vitamin C solution.

[0054] (5) Fresh culture water and sample solution were replaced every 24 hours, and the administration period was 2 days.

[0055] (6) After the administration period, the experimental zebrafish samples were processed as follows: the zebrafish were frozen at -20°C, weighed and cut into pieces for later use, an appropriate amount of pre-cooled PBS was added at a weight-to-volume ratio of 1 g:9 mL, and the mixture was thoroughly homogenized with a glass homogenizer in an ice bath. The homogenate was aspirated into a centrifuge tube, centrifuged at 5000 × g for 5 min at 5°C, the supernatant was collected, and stored at -20°C.

[0056] (7) According to the instructions of the ELISA kit, the 5α-reductase activity, loricrin content, and hyaluronic acid content in zebrafish were determined.

[0057] The results are shown in Table 11. Niacinamide, tea polyphenols and vitamin C were selected, and their oil control effects were evaluated by this method. The results showed that niacinamide had a comprehensive score of 7 points, which was determined to have a good oil control effect. The comprehensive score of tea polyphenols was 4 points, which was determined to have a general oil control effect. The comprehensive score of vitamin C was 0 points, which was determined to have no oil control effect.

[0058] Table 11: Sample test results Sample name Niacinamide Tea polyphenols Vitamin C Sample concentration (mg / mL) 0.5 1.0 0.1 5α-reductase activity (U / mL) 27.46 31.55 36.52 Scoring 4 2 0 Loricrin content (ng / mL) 6.14 6.23 5.29 Scoring 2 2 0 Hyaluronic acid content (ng / mL) 60.23 56.17 54.26 Scoring 1 0 0 Total score 7 4 0 Oil control effect good generally none The raw materials and equipment used in the present invention, unless otherwise specified, are all commonly used raw materials and equipment in the art; the methods used in the present invention, unless otherwise specified, are all conventional methods in the art.

[0059] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent transformation made to the above embodiment based on the technical essence of the present invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. A rapid evaluation method for the oil control efficacy of cosmetic raw materials, characterized in that include: 1) Selecting a number of zebrafish and dividing them into a blank group and at least one sample group; the blank group is incubated with aquaculture water, and the sample group is incubated with a sample solution containing the cosmetic raw material to be tested; 2) After the incubation, the 5α-reductase activity, loricrin content, and hyaluronic acid content in zebrafish of each group were measured; 3) Score the oil control efficacy of the tested cosmetic raw materials based on the test results in 2).

2. The rapid evaluation method according to claim 1, characterized in that: 3) In the oil control effect scoring criteria, the oil control effect scoring criteria are as follows: The sum of the three scores is recorded as the total score. 6-8 points are considered to be good oil control effect, 3-5 points are considered to be fair oil control effect, and 2 points or less are considered to be no oil control effect.

3. The rapid evaluation method according to claim 1, characterized in that: 1), the incubation temperature is 25-30°C, fresh aquaculture water and sample solution are replaced every 20-30 hours, and the incubation time is 40-60 hours.

4. The rapid evaluation method according to any one of claims 1 to 3, characterized in that: 1), the zebrafish is a 72 hpf wild-type AB strain zebrafish fry with normal development and consistent status.

5. The rapid evaluation method according to claim 4, characterized in that: 1), the method for obtaining the 72hpf wild-type AB strain zebrafish fry is as follows: sexually mature wild-type AB strain zebrafish are raised in separate tanks in a zebrafish circulation system, with a water temperature of 26±2°C; a pH value of 7-7.5; an electrical conductivity of 500-550μs / cm; a light / dark cycle of (12-16)h / (8-12)h, and a light intensity of 80-100W; the males and females are paired at a ratio of 1:(1.5-2.5) one day before mating, and natural mating and spawning are carried out, and the eggs are transferred into culture water to cultivate 72hpf wild-type AB strain zebrafish fry.

6. The rapid evaluation method according to any one of claims 1 to 3, characterized in that: 1), the number of zebrafish in each group was 20 to 30, and multiple parallel groups were set up in each group.

7. The rapid evaluation method according to any one of claims 1 to 3, characterized in that: 1), the concentration of the cosmetic raw material to be tested in the sample solution is not higher than its maximum safe concentration, and the amount of each group of sample solutions is 3 to 5 mL.

8. The rapid evaluation method according to claim 7, characterized in that: 1), the standard of the maximum safe concentration of the cosmetic raw material to be tested in the sample solution is: the highest concentration at which more than 80% of the zebrafish fry after 3 days of cultivation do not show any mortality or other toxic effects.

9. The rapid evaluation method according to any one of claims 1 to 3, characterized in that: 1), the amount of the aquaculture water used in each group is 3-5 mL.

10. The rapid evaluation method according to any one of claims 1 to 3, characterized in that: 2) Specifically comprising: freezing the zebrafish, cutting them into pieces, adding pre-cooled PBS, fully homogenizing, centrifuging the resulting homogenate, collecting the supernatant, and freezing and storing; using ELISA to determine the 5α-reductase activity, loricrin content, and hyaluronic acid content in each group of zebrafish.

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