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.
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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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