Method for evaluating eye stimulation of infant cosmetics by using zebra fish
A zebrafish and cosmetic technology, applied in the field of toxicity evaluation, can solve the problems of non-compliance with animal protection and welfare, subjectivity of test scores, long experimental period, etc., to achieve eye irritation, easy microscopic manipulation, and high accuracy Effect
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Embodiment 1
[0038] The selection of the safety concentration of baby cosmetics to be tested in embodiment 1:
[0039] 1) The present invention selects the test model as the wild type AB;
[0040] 2) Using dimethyl sulfoxide (DMSO) as a solvent to prepare infant cosmetics containing 0.59% acyl glutamate into a high-concentration mother solution with a concentration of 120 mg / mL, and store at -20°C;
[0041]3) Dilute the prepared mother solution with embryo culture solution into working solutions of 10, 20, 40, 60, 80, 100 and 120 μg / mL. The solutions are prepared and used immediately, and the concentration of DMSO is kept less than one thousandth;
[0042] 4) Inject the culture solution containing the baby cosmetics to be tested and the culture solution not containing the sample to be tested into a 6cm petri dish, and control the volume of the culture solution at 8-9mL.;
[0043] 5) Select healthy and well-developed embryos reproduced and developed by zebrafish for exposure experiments, a...
Embodiment 2
[0047] Example 2 In Vivo Observation of Eye Damage of Cosmetic Zebrafish Containing 0.59% Acyl Glutamate Content
[0048] 1) Randomly select 3dpf zebrafish with normal development, 20 in each group;
[0049] 2) Select cosmetics containing 0.59% acyl glutamate content, and other components are mild and non-irritating, with a concentration of 10 μg / mL and 20 μg / mL as the experimental group, and 0.1% DMSO as the control group.
[0050] 3) Place the petri dish in a constant temperature light-controlled incubator, the temperature is controlled at 28±1°C, the light and dark cycle is 14 / 10h, the semi-static culture method is adopted, the culture medium is replaced every 24h, and the exposure time is 72h;
[0051] 4) Observe the morphological changes of the eyes of zebrafish juveniles under a stereomicroscope and take pictures to record;
[0052] 5) Measure iris bandwidth and pupil diameter with Image J software, and calculate the ratio of the two.
[0053] The result is as image ...
Embodiment 3
[0054] Example 3 Analysis of Behavioral Trajectories of Zebrafish After Exposure to Cosmetics Containing 0.59% Acyl Glutamate Content for 72h
[0055] 1) Randomly select 3dpf zebrafish with normal development, 20 in each group;
[0056] 2) Choose a cosmetic containing 0.59% of acyl glutamate, and other components are mild and non-irritating, with a concentration of 10 μg / mL and 20 μg / mL as the experimental group, and 0.1% DMSO as the control group;
[0057] 3) Place the petri dish in a constant temperature light-controlled incubator, the temperature is controlled at 28±1°C, the light and dark cycle is 14 / 10h, the semi-static culture method is adopted, the culture medium is replaced every 24h, and the exposure time is 72h;
[0058] 4) After 72 hours, wash 3 times, place in a 96-well plate, one fish per well, 15 fish per group, put them into the zebrafish behavior track tracking system (zebrafish behavior detector) and let stand for 10 minutes;
[0059] 5) Immediately give ligh...
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