Whitening effect evaluation method based on small fish embryos
A technology of whitening effect and evaluation method, which is applied in biochemical equipment and methods, microbial determination/inspection, color/spectral property measurement, etc. Whitening effect and other issues, to achieve the effect of short experimental period, high-efficiency raw material efficacy screening and activity evaluation, and low cost
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Embodiment 1
[0045] The determination of embodiment 1 detection wavelength
[0046] (1) Select healthy zebrafish fertilized eggs under a microscope and place them in a petri dish containing 10 mL of Holtfreter’s Solution, with no more than 300 embryos per dish and a culture density of no more than 10 embryos / cm 2 , to avoid death caused by too high density, and then cultured in a constant temperature incubator at 28°C to 24hpf;
[0047] (2) Transfer the healthy embryos developed to 24hpf in step (1) to a 24-well plate containing 500 μL Holtfreter's Solution, 20 embryos per well; then add the positive compound 1-benzene to each well of the 24-well plate Base-2-thiourea (Phenylthiourea, PTU) (final concentration: 30 μg / mL), incubating at 28°C in a constant temperature incubator until 48hpf, using no positive compound as the blank group;
[0048] (3) Transfer 20 embryos cultured to 48hpf in step (2) to a centrifuge tube and process in a tissue homogenizer at 45Hz for 30s to obtain a homogena...
Embodiment 2
[0051] Example 2 Evaluation of the whitening effect of α-arbutin
[0052] (1) At about 3:30 pm, select 4 pairs of healthy zebrafish and place them in mating boxes, with 2 females and 2 males in each box, and separate the males and females with partitions; at 9:00 am the next day, remove the partitions , collect the embryos after 40 minutes;
[0053] (2) Select healthy fertilized eggs under a microscope and place them in a petri dish containing 10mL Holtfreter’s Solution, with no more than 300 embryos per dish and a culture density of no more than 10 embryos / cm 2 , to avoid death caused by too high density, and then cultured in a constant temperature incubator at 28°C to 24hpf;
[0054] (3) Transfer the healthy embryos developed to 24hpf in step (2) to a 24-well plate containing 500 μL Holtfreter'sSolution, 30 embryos per well; then add α-arbutin to each well of the 24-well plate, Set the final concentrations to 100mM, 10mM and 1mM respectively, set up 3 replicate wells for e...
Embodiment 3
[0058] Example 3 Evaluation of the whitening efficacy of phenylethyl resorcinol (Phenylethyl Resorcinol, PHR) and ascorbic acid ethyl ether (3-O-Ethyl Ascorbicacid, VCE)
[0059] (1) At about 3:30 pm, select 4 pairs of healthy zebrafish and place them in mating boxes, with 2 females and 2 males in each box, and separate the males and females with partitions; at 9:00 am the next day, remove the partitions , collect the embryos after 40 minutes;
[0060] (2) Select healthy fertilized eggs under a microscope and place them in a petri dish containing 10mL Holtfreter’s Solution, with no more than 300 embryos per dish and a culture density of no more than 10 embryos / cm 2 , to avoid death caused by too high density, and then cultured in a constant temperature incubator at 28°C to 24hpf;
[0061] (3) Transfer the healthy embryos developed to 24hpf in step (2) to a 24-well plate containing 600 μL Holtfreter's Solution, 30 embryos per well; then in each well of the 24-well plate, add P...
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