Antioxidant with function of inhibiting melanin production as well as preparation method and use of antioxidant
A technology for inhibiting melanin and anti-oxidants, applied in skin care preparations, functions and applications of food ingredients, etc., to achieve broad application prospects, maintain activity, and simple steps
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Embodiment 1
[0033]In the present embodiment, the preparation method of an antioxidant capable of inhibiting melanin production is as follows:
[0034] 1. Pulverize the Morchella fruiting bodies and pass through an 80-mesh sieve to obtain powder;
[0035] 2. Add 100 g of the obtained powder in sequence to petroleum ether and 95vt% ethanol and reflux at 60°C for 2 hours, repeat three times, and dry the obtained precipitate for later use;
[0036] 3. Take 100 g of the precipitate obtained in step 2, add it into distilled water at a liquid-to-solid ratio of 22 mL / g, and conduct ultrasonic-assisted extraction. The ultrasonic power is 300 W, and the ultrasonic time is 30 min. Repeat the extraction 3-4 times, and combine the extracts;
[0037] 4. Concentrate the extract obtained in step 3 to 200mL, add four times the volume of 95vt% ethanol, let stand at 4°C for 10h, centrifuge, and collect the precipitate;
[0038] 5. Dissolve the precipitate obtained in step 4 with distilled water, then add a...
Embodiment 2
[0041] Embodiment 2: In vitro free radical scavenging ability test
[0042] 1. Hydroxyl radical scavenging experiment
[0043] Accurately prepare morel polysaccharide FMP solution (25, 50, 100, 200, 300 and 400μg / mL), 6mM ferrous sulfate solution, 2.4mM hydrogen peroxide solution, 6mM ethanol-salicylic acid solution, each take 1mL sulfuric acid Ferrous solution, 1mL ethanol-salicylic acid solution, add 1mL polysaccharide sample solution, oscillate and mix well and let stand for 10min. Add 1 mL of hydrogen peroxide solution and react for 30 min. Ascorbic acid was used as a positive control, and the reaction was carried out in a constant temperature water bath at 37°C for 30 minutes and protected from light. Absorbance at 510nm with a UV-Vis spectrophotometer. Deionized water was used instead of polysaccharide solution as a blank group, and instead of salicylic acid as a control group. Three parallel groups were set up for each experiment. Calculation of hydroxyl radical sca...
Embodiment 3
[0048] Embodiment 3: In vitro inhibition of melanin activity test
[0049] 1. Determination of FMP polysaccharides on the proliferation of B16F10 cells
[0050] MTT method was used to measure the effect of samples on the proliferation of B16F10 cells, and the concentration of passaged cells was adjusted to 5×10 3 After each well, add 96-well culture plate, 100 μL per well; culture for 24 hours, absorb the culture medium after the cells adhere to the wall, replace the medium once with fresh culture medium treated with different drug concentrations, set up 8 replicate wells in each group, and place At 37°C, 5% CO 2 Cultivate under conditions for 48 hours; add MTT solution (5 mg / mL, 20 μL / well) 4 hours before the end; discard the supernatant after 48 hours, add 100 μL dimethyl sulfoxide to each well, shake gently for 5 minutes to fully dissolve the crystals; then The 96-well plate was placed in a microplate reader, and the absorbance was measured at a wavelength of 570 nm. The...
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