Procyanidine with melanogenesis inhibiting activity and preparation method and application thereof
A technology for inhibiting melanin and proanthocyanidins, which is applied in the directions of organic active ingredients, medical preparations containing active ingredients, and organic chemistry to achieve the effects of stable properties, convenient extraction and easy storage.
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Embodiment 1
[0023] Example 1: Preparation of procyanidins with melanin production-inhibiting activity
[0024] 1.1 Extraction and separation
[0025] The lychee husk (50g) was added with a volume percentage of 60% ethanol aqueous solution according to the mass-to-liquid ratio of 1:5g / ml, heated and refluxed for 2 hours at 60°C, and the supernatant was collected; the supernatant was heated at 45-50°C. Concentrate under reduced pressure to ethanol-free, dilute the concentrate with distilled water and pass it through an AB-8 macroporous adsorption resin column, and elute with distilled water and 90% ethanol aqueous solution by volume respectively; The solution was concentrated under reduced pressure at 45-50 °C to obtain the total extract; the total extract was separated by an ODS-A reversed-phase medium pressure column at a flow rate of 10 mL / min, and methanol water with a concentration of 25% and 40% by volume was used respectively. Elution, collect 40% methanol water eluted part and conc...
Embodiment 2
[0030] Example 2: Determination of melanin production-inhibiting activity of proanthocyanidins
[0031] 1. Instruments and materials
[0032] A375 human melanoma cells were purchased from ATCC, DMEM medium, fetal bovine serum (FBS), 0.25% trypsin-EDTA, penicillin + streptomycin (double antibody), phosphate buffer were purchased from Thermo Fisher Scientific, USA, β -Arbutin and dimethyl sulfoxide (DMSO) were purchased from Sigma Company in the United States.
[0033] 2. Evaluation of melanin production inhibition activity
[0034] A375 human melanoma cells in good growth state were plated at 6 × 10 5 Cells / well were inoculated in a 6-well culture plate, and after culturing for 12 hours, after the cells adhered to the wall, they were replaced with 2 ml of 1% BSA serum-free medium containing different concentrations of β-arbutin, grape seed proanthocyanidin or LPOPC, and continued to culture for 24 hours. , washed twice with pH 7.4 phosphate buffer, add trypsin-EDTA to digest...
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