Method for evaluating skin irritation of cosmetics

Abstract

The invention relates to the technical field of cosmetic safety evaluation, and discloses a method for evaluating skin irritation of cosmetics aiming at the problem of high subjectivity of evaluationindexes of a cosmetic irritation evaluation method. The method comprises the following steps of setting a normal control group and a test group, and adding the to-be-tested cosmetics into the test group in a water-soluble manner, taking zebra fishes in a normal control group and a test group, placing the zebra fishes under a fluorescence microscope for photographing, storing pictures, counting thenumber of neutrophils on the skin of the zebra fishes, and performing statistical analysis. According to the evaluation method, evaluation index selection, obtaining and processing analysis are moreobjective, the evaluation result can be quantified, whether a single cosmetic has irritation or not can be evaluated, and the irritation strength of different cosmetics can be compared; due to the characteristics of low zebra fish feeding cost, small size, short development cycle and high single spawning frequency, multiple cosmetics can be evaluated at the same time, the evaluation efficiency ishigh, the evaluation cost is low, and laboratory evaluation is facilitated.

Description

technical field

[0001] The invention relates to the technical field of cosmetic safety evaluation, in particular to a method for evaluating skin irritation of cosmetics. Background technique

[0002] At present, the common methods for evaluating the skin irritation of cosmetics include closed patch test on human body, animal experiment method, chicken chorioallantoic membrane test method and three-dimensional reconstructed skin model, etc.

[0003] Closed patch test on human body: Select 30 eligible subjects aged 18-60, all without skin inflammation and other diseases or conditions that are not suitable for the test. The test substance was prepared into two concentration levels of 1% and 5% mass fraction with deionized water. Add 0.020-0.025mL test sample dropwise on the filter paper attached to the spot tester, place the filter paper in the spot tester, set solvent holes and blank control holes. Stick the spot tester containing the sample on the subject's back with non-ir...

Examples

Embodiment 1

[0037] A method for evaluating skin irritation of cosmetics, wherein the mass content of glycerin in the cosmetics is 10%, and other components are mild and non-irritating, comprising the following steps:

[0038] (1) Determination of safe concentration: Place the transgenic neutrophil green fluorescent zebrafish 2 days after fertilization in a water-containing orifice plate with a water capacity of 3ml and 30 zebrafish, and add different concentrations of zebrafish to different orifice plates in a water-soluble manner. The concentration of the cosmetic to be tested is 1.39, 2.78, 5.56, 11.1, 22.2, 44.4 μL / mL. After 18 hours of culture, the death and organ damage of the zebrafish are counted. The results are shown in Table 1, so as to determine the safe concentration of the cosmetic The upper limit is 11.1 μL / mL;

[0039] (2) Set up a normal control group: 30 transgenic neutrophil green fluorescent zebrafish 2 days after fertilization were used as test subjects, placed in an o...

Embodiment 2

[0046] A method for evaluating the skin irritation of cosmetics, the content of methylisothiazolinone in the cosmetics is 70ppm, and the other components are all mild and non-irritating, comprising the following steps:

[0047] (1) Determination of safe concentration: Place the transgenic neutrophil green fluorescent zebrafish 2 days after fertilization in a water-containing orifice plate with a water capacity of 3ml and 30 zebrafish, and add different concentrations to different orifice plates in a water-soluble manner. The concentrations of the cosmetics to be tested were 1.39, 2.78, 5.56, 11.1, 22.2, and 44.4 μL / mL in turn. After 18 hours of culture, the death and organ damage of the zebrafish were counted. The results are shown in Table 1, so as to determine the safety of the cosmetics The upper limit of concentration is 5.56 μL / mL;

[0048] (2) Set up a normal control group: 30 transgenic neutrophil green fluorescent zebrafish 2 days after fertilization were used as test ...

Embodiment 3

[0055] A method for evaluating the skin irritation of cosmetics, the content of ethanol in the cosmetics is 3%, and the other components are mild and non-irritating, comprising the following steps:

[0056] (1) Determination of safe concentration: Place the transgenic neutrophil green fluorescent zebrafish 2 days after fertilization in a water-containing orifice plate with a water capacity of 3ml, 30 zebrafish, and add different concentrations to different orifice plates in a water-soluble manner. The concentrations of the cosmetics to be tested were 1.39, 2.78, 5.56, 11.1, 22.2, and 44.4 μL / mL in turn. After 18 hours of culture, the death and organ damage of the zebrafish were counted. The results are shown in Table 1, so as to determine the safety of the test. The upper limit of concentration is 22.2 μL / mL;

[0057] (2) Set up a normal control group: 30 transgenic neutrophil green fluorescent zebrafish 2 days after fertilization were used as test subjects, placed in an orifi...

technical field

[0001] The invention relates to the technical field of cosmetic safety evaluation, in particular to a method for evaluating skin irritation of cosmetics. Background technique

[0002] At present, the common methods for evaluating the skin irritation of cosmetics include closed patch test on human body, animal experiment method, chicken chorioallantoic membrane test method and three-dimensional reconstructed skin model, etc.

[0003] Closed patch test on human body: Select 30 eligible subjects aged 18-60, all without skin inflammation and other diseases or conditions that are not suitable for the test. The test substance was prepared into two concentration levels of 1% and 5% mass fraction with deionized water. Add 0.020-0.025mL test sample dropwise on the filter paper attached to the spot tester, place the filter paper in the spot tester, set solvent holes and blank control holes. Stick the spot tester containing the sample on the subject's back with non-ir...

Examples

Embodiment 1

[0037] A method for evaluating skin irritation of cosmetics, wherein the mass content of glycerin in the cosmetics is 10%, and other components are mild and non-irritating, comprising the following steps:

[0038] (1) Determination of safe concentration: Place the transgenic neutrophil green fluorescent zebrafish 2 days after fertilization in a water-containing orifice plate with a water capacity of 3ml and 30 zebrafish, and add different concentrations of zebrafish to different orifice plates in a water-soluble manner. The concentration of the cosmetic to be tested is 1.39, 2.78, 5.56, 11.1, 22.2, 44.4 μL / mL. After 18 hours of culture, the death and organ damage of the zebrafish are counted. The results are shown in Table 1, so as to determine the safe concentration of the cosmetic The upper limit is 11.1 μL / mL;

[0039] (2) Set up a normal control group: 30 transgenic neutrophil green fluorescent zebrafish 2 days after fertilization were used as test subjects, placed in an o...

Embodiment 2

[0046] A method for evaluating the skin irritation of cosmetics, the content of methylisothiazolinone in the cosmetics is 70ppm, and the other components are all mild and non-irritating, comprising the following steps:

[0047] (1) Determination of safe concentration: Place the transgenic neutrophil green fluorescent zebrafish 2 days after fertilization in a water-containing orifice plate with a water capacity of 3ml and 30 zebrafish, and add different concentrations to different orifice plates in a water-soluble manner. The concentrations of the cosmetics to be tested were 1.39, 2.78, 5.56, 11.1, 22.2, and 44.4 μL / mL in turn. After 18 hours of culture, the death and organ damage of the zebrafish were counted. The results are shown in Table 1, so as to determine the safety of the cosmetics The upper limit of concentration is 5.56 μL / mL;

[0048] (2) Set up a normal control group: 30 transgenic neutrophil green fluorescent zebrafish 2 days after fertilization were used as test ...

Embodiment 3

[0055] A method for evaluating the skin irritation of cosmetics, the content of ethanol in the cosmetics is 3%, and the other components are mild and non-irritating, comprising the following steps:

[0056] (1) Determination of safe concentration: Place the transgenic neutrophil green fluorescent zebrafish 2 days after fertilization in a water-containing orifice plate with a water capacity of 3ml, 30 zebrafish, and add different concentrations to different orifice plates in a water-soluble manner. The concentrations of the cosmetics to be tested were 1.39, 2.78, 5.56, 11.1, 22.2, and 44.4 μL / mL in turn. After 18 hours of culture, the death and organ damage of the zebrafish were counted. The results are shown in Table 1, so as to determine the safety of the test. The upper limit of concentration is 22.2 μL / mL;

[0057] (2) Set up a normal control group: 30 transgenic neutrophil green fluorescent zebrafish 2 days after fertilization were used as test subjects, placed in an orifi...

Images