Application of melia azedarach leaf extract in whitening product
A technology of neem leaves and extracts is applied in the application field of neem leaf extracts in whitening products, and achieves the effects of whitening development and application value, inhibitory activity, and good development and application value.
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Embodiment 1
[0016] The preparation method of neem leaf extract is as follows: the neem leaf is pulverized, placed in supercritical CO 2 In the extraction device, the temperature of the extraction tank is controlled to be 50°C, the pressure is 40Mpa, the temperature of the first separation tank is 40°C, the pressure is 5Mpa, the temperature of the second separation tank is 35°C, the pressure is 4Mpa, CO 2 The flow rate is 32L / h, extract for 2 hours to obtain medicinal residues, add 75% ethanol to the medicinal residues, the volume mass ratio of ethanol and medicinal residues is 8mL: 1g, extract twice at 60°C for 1 hour each time, combine the leaching The extract was concentrated under reduced pressure at 70°C and 0.2 bar, and freeze-dried to obtain the obtained product.
Embodiment 2
[0018] Neem leaf extract inhibits tyrosinase activity experiment:
[0019] Take 5 test tubes, wherein No. 1 and No. 2 test tubes add 1 mL of 0.05wt% neem leaf extract solution, No. 3 and No. 4 test tubes add 1 mL of phosphate buffer (pH=6.8) respectively, 1 mL of 0.05 wt% arbutin solution. Add 0.5 mL of tyrosinase solution (enzyme activity unit 125u / mL) to No. 1, No. 3 and No. 5 test tubes, add 0.5 mL of phosphate buffer (pH=6.8) to No. 2 and No. 4 test tubes, and 5 Shake the test tubes evenly, put them in a water bath at 35°C for 10 minutes, add 2 mL of 0.05 wt% tyrosine solution to it, react for 10 minutes, measure the absorbance T of each test tube at 475 nm, and mark them as T1, T2, T3, T4, T5, And calculate the inhibition rate.
[0020] The calculation formula of inhibition rate is: I(%)=[1-(T1-T2) / (T3-T4)]×100%
[0021] It is calculated that the inhibition rate of 0.05wt% neem leaf extract solution to tyrosinase can reach 81.1%, while the inhibition rate of arbutin as...
Embodiment 3
[0023] Inhibitory effect of neem leaf extract on melanin:
[0024] The improved method of Hosoi et al. was used to determine the inhibitory effect of neem leaf extract on melanin. B16 melanocytes (purchased from Shanghai Yansheng Industrial Co., Ltd.) were cultured in a 96-well plate at a density of 1 × 105. After 24 hours, the medium was changed, and 0.05 wt% neem leaf extract solution was added. After 72 hours, the cells were incubated with phosphate buffer ( pH = 6.8) and washed twice, the sample was air-dried, dissolved in 200 μL 1mol / L NaOH (containing 1% DMSO), cooled after heating at 80°C to 1h, and read the absorbance value on an enzyme-linked immunosorbent assay with a wavelength of 475nm. Calculate the inhibition rate; use the same method to measure the absorbance of 0.05wt% arbutin and calculate the inhibition rate.
[0025] It is calculated that the inhibition rate of melanin by 0.05wt% neem leaf extract solution can reach 69.8%, while the inhibition rate of arbut...
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