Method for evaluating mildness and irritation of cosmetics by using zebra fish embryos

A technology of cosmetics and zebrafish, which is applied in the field of mild irritation evaluation of cosmetics, which can solve the problems of subjective factors, inability to accurately distinguish irritation levels, animal damage, etc.

Inactive Publication Date: 2021-05-28
广州鲁比生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the above method, the eye irritation test of rabbits requires the use of New Zealand rabbits for the eye irritation test, but this method has a long test period and high cost, and there are still some differences between the animal test results and the human body, so it is not suitable for the current cosmetics. The trend and requirements of animal substitution experiments, and the greater harm to animals, this method has received more and more criticism and condemnation; the turbidity experiment of the bull's eye cornea uses isolated organs for experiments, and the use of isolated bovine cornea , directly contact the substance to be tested, and classify the irritation of the substance to be tested by measuring the turbidity and permeability of the bovine cornea and calculating the formula
This method can be used to evaluate whether cosmetics are irritating, but it cannot accurately distinguish the irritating level, and the source of raw materials is often limited when used on a large scale; the chicken chorioallantoic membrane experiment is to use the chicken embryo chorioallantoic membrane and eye Similar characteristics of conjunctival tissue organization, assessed by observing the stimuli response (bleeding, angiolysis, coagulation) of blood vessels after exposure of the allantoic membrane to the test substance
The experiment period is often long, and the experiment results are easily affected by the subjective factors of the experimenter, so it is difficult to quantify, and there are many cases of false positives or false negatives in the experimental results (Wang Beiming, Chen Yuxuan, Zhong Minxian. Research progress on cosmetic irritation evaluation methods[J] .Flavor, Fragrance and Cosmetics, 2017, 000(005):69-74)
At present, there is no efficient method for mild irritation evaluation of cosmetics

Method used

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  • Method for evaluating mildness and irritation of cosmetics by using zebra fish embryos
  • Method for evaluating mildness and irritation of cosmetics by using zebra fish embryos
  • Method for evaluating mildness and irritation of cosmetics by using zebra fish embryos

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039]The Basic Method of Mild Stimulus Experiment

[0040] Fish embryo collection: Put the zebrafish into the fish embryo spawning box 24 hours in advance for mating and spawning. After fertilization, fish embryos are collected with a fish embryo collector, placed in a glass plate, placed in a biochemical incubator, and set aside.

[0041] Exposure test: take out 24hpf, 48hpf, 72hpf fish embryos, use 6-well plate or 24-well plate as the test container, put 5-50 fish embryos into each well, and add 0.5-5mL of the solution to be tested. A total of 3 parallel groups were repeated. Zebrafish system culture water or distilled water or PBS was used as blank control, and SDS (sodium dodecyl sulfate) or AES (sodium fatty alcohol polyoxyethylene ether sulfate) or sodium hydroxide was used as positive control. Observe under a stereomicroscope. In 0.5 ~ 10min, observe and video record.

[0042] Data collection and processing: fish embryo death or coagulation rate, based on which the...

Embodiment 2

[0045] Mild irritation test on cosmetic raw materials (SDS strong irritation)

[0046] 1. Experimental method: AB line zebrafish embryos with normal development of 24 hours were selected for the experiment. A 24-well plate was used as the experimental container, 20 embryos were placed in each well, and 1 mL of the test solution was added. A total of 3 parallel groups were repeated.

[0047] 2. Data statistics: After adding the test solution, observe the stimulation response of fish embryos within 5 minutes and 10 minutes under a stereomicroscope, and record them.

[0048] 3. Experimental results and judgment of the strength of irritation (Table 2-1 and Table 2-2):

[0049] Table 2-1 Experimental Results and Stimulant Judgment

[0050]

[0051] Note: The number of fish embryo death (coagulation) refers to the total sum of fish embryo death and coagulation.

[0052] Table 2-2 Experimental results and the judgment of the intensity of irritation

[0053]

[0054]

[0...

Embodiment 3

[0060] Mild irritation test on cosmetic raw materials (AES strong irritation)

[0061] 1. Experimental method: AB line zebrafish embryos with normal development of 24 hours were selected for the experiment. A 24-well plate was used as the experimental container, 20 embryos were placed in each well, and 1 mL of the test solution was added. A total of 3 parallel groups were repeated.

[0062] 2. Data statistics: After adding the test solution, observe the stimulation response of fish embryos within 5 minutes and 10 minutes under a stereomicroscope, and record them.

[0063] 3. Experimental results and judgment of the strength of irritation (Table 3-1 and Table 3-2):

[0064] Table 3-1 Experimental Results and Judgment of Stimulant Strength

[0065]

[0066]

[0067] Note: The number of fish embryo death (coagulation) refers to the total sum of fish embryo death and coagulation.

[0068] Table 3-2 Experimental Results and Judgment of Irritant Strength

[0069]

[007...

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Abstract

The invention relates to a method for evaluating mildness and irritation of cosmetics by applying zebra fish embryos, and belongs to the technical field of cosmetic evaluation. According to the method, cosmetics to be detected and zebra fish embryos of 20-72 hpf are mixed, the stimulation condition of the fish embryos is observed and counted, and mild stimulation evaluation of the cosmetics is achieved. The observation indexes comprise fish embryo fishtailing and fish embryo death or coagulation. According to the method, the zebra fish embryos are used as experimental carriers, and the zebra fish embryos are transparent, so that observation is facilitated; the egg laying amount is large, the number of experimental samples is large, and repeatability is high; and rapid response to external irritation can be made. According to the method, the mildness and irritation of the zebra fish embryos to cosmetics are evaluated, experimental samples can be screened quickly and scientifically with high throughput, and the research and development process of cosmetics related enterprises is accelerated. And meanwhile, more evaluation choices are provided for cosmetic-related enterprises and consumers to select safe cosmetic raw materials and cosmetics.

Description

technical field [0001] The invention relates to the technical field of cosmetic evaluation, in particular to a method for evaluating mild irritation of cosmetics using zebrafish embryos. Background technique [0002] The mild irritation of cosmetics is of great significance to the formulation research, production and marketing of cosmetics. At present, the main methods used include rabbit eye test, bull's eye test and chicken embryo allantoic membrane test to evaluate the mild irritation of cosmetics. "Chicken chorioallantoic membrane test for in vitro eye irritation evaluation of cosmetics" reported the evaluation method of using chicken chorioallantoic membrane to evaluate the eye irritation of cosmetics in vitro. "TSHRH 011-2018 Cosmetics Eye Irritation Test Chicken Chorioallantoic Membrane Test" adopts the methods of chicken chorioallantoic membrane test and eye irritation test. In the above method, the eye irritation test of rabbits requires the use of New Zealand rabb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/00
CPCG01N33/00G01N2033/0078
Inventor 刘云啸周淑媛黄晓婷
Owner 广州鲁比生物科技有限公司
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