New application of B Salvianolic acid taken as cosmetics additives with functions of whitening skin, removing freckle and resisting wrinkle
A technology of salvianolic acid and food additives, which is used in the field of salvianolic acid as a cosmetic additive for whitening, freckle and anti-wrinkle, and can solve problems such as economic losses
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Embodiment 1
[0018] Accurately weigh 0.05g of L-dopa, dissolve it with 0.1mol / L PBS (pH6.8), dilute to 10mL, keep away from light, and dissolve with ultrasonic. L-dopa solution is easy to oxidize and should be prepared immediately.
[0019] Dissolve 25KU of tyrosinase powder with 0.1mol / L PB (pH6.8) and make the volume to 250mL. Dispense the prepared tyrosinase solution into centrifuge tubes, seal and place in a -20°C refrigerator save.
[0020] Accurately weigh 10.00 mg of the total phenolic acid sample of Salvia miltiorrhiza, and dissolve it in 10 mL of double-distilled water to obtain a salvianolic acid solution. Three concentration groups of 1.0 mg / mL, 0.1 mg / mL and 0.01 mg / mL of the extract were finally obtained through stepwise dilution with double distilled water. Store at 4°C for later use.
[0021] The frozen tyrosinase solution was placed in a 37°C water bath, and the solution in the tube was quickly thawed by shaking quickly. The thawed tyrosinase solution must be used up wit...
specific Embodiment 2
[0035] Effect of salvianolic acid on tyrosinase activity of A375 and Hacat co-cultured cells
[0036] The A375 cells and Hacat cells in the logarithmic growth phase were respectively digested with 0.25% trypsin, centrifuged, and suspended in MEM culture medium. Mix the two cell suspensions at a ratio of 1:2, and adjust the cell density to 1.5×10 4 cells / mL inoculated in a 96-well plate, 200 μL per well, at 37°C and 5% CO 2 Incubate for 24h. According to the results of the MTT experiment, select 4 concentrations (400 μg / mL, 200 μg / mL, 100 μg / mL, 50 μg / mL) of salvianolic acid in the inverted culture medium, 200 μL per well, set 6 duplicate wells in each group, and continue to incubate for 48 hours , washed twice with PBS, added 100 μL of 1% Triton-X solution to each well, quickly placed in a -80°C refrigerator for 30 minutes, and then thawed at room temperature after removal to fully lyse the cells. Add 50 μL of 0.5% L-DOPA solution to each well, place the culture plate on a ...
specific Embodiment 3
[0041] Effects of Salvianolic Acid on Melanin Synthesis in A375 and Hacat Co-cultured Cells
[0042] Mix A375 cells and Hacat cells at a ratio of 1:2, and adjust the cell density to 4.5×10 4 cells / mL inoculated in a 6-well plate, 2 mL per well, at 37°C and 5% CO 2Incubate for 24h. Discard the culture medium, select 4 concentrations (400 μg / mL, 200 μg / mL, 100 μg / mL, 50 μg / mL) of total salvianolic acid according to the results of the MTT experiment, 2 mL per well, continue to incubate for 48 h, remove the culture medium, and wash with PBS 2 times, discard the culture medium, add 0.25% trypsin 1mL / well to digest, then add 1mL MEM culture medium to stop the digestion, make a cell suspension, collect the cell suspension into a 15mL centrifuge tube, centrifuge, discard the supernatant, Add 100 μL NaOH with a concentration of 1mol / L, and bathe in water at 37°C for 1 hour to fully lyse the cells and dissolve the melanin granules. Add 400 μL double-distilled water to dilute, mix tho...
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