Diffractive ring mangiferin in mango leaf and new application of mango leaf extract containing diffractive ring mangiferin
A technology of mangiferin and mango leaves, applied in the field of mango leaf extracts, can solve the problems of inducing cancer and not being widely used, and achieve the effects of preventing wrinkles, strong antioxidant activity, and preventing or treating skin diseases
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Embodiment 1
[0031] Preparation of Active Ingredient Monomer
[0032] 5 kg of mango leaves were reflux extracted twice with 9 times the amount of ethanol, each time for 3 hours, the filtrates were combined, and the solvent was recovered; dried in a vacuum oven at 50°C to obtain 1.163 kg of extract.
[0033] Take 600g of the above-mentioned extract, add 5L of distilled water to dissolve, extract three times with 5L of ethyl acetate to obtain ethyl acetate layer and water layer respectively; the water layer part contains a large amount of precipitation, take 4 / 5 of the volume of the suspension in the water layer, and concentrate under reduced pressure to 2L, centrifuged, filtered, and the supernatant was treated with a macroporous adsorption resin (H2O→95%EtOH) to obtain 84g of 95% ethanol eluate.
[0034] Take 72 g of the above-mentioned 95% ethanol eluate and perform silica gel column chromatography [chloroform-methanol (10:1 → 5:1) → chloroform-methanol-water (7:3:1 → 6:4:1, lower layer) ...
Embodiment 2
[0036] Confirmation of the structure of the active ingredient
[0037] The structure of the two compounds obtained in Example 1 (i.e., split-mangiferin A and split-ring mangiferin B) was confirmed (entrusted to Tianjin WuXi Kangde New Drug Development Co., Ltd. Analysis and Testing Center, 400MHz NMR), and the analysis results showed The structures of these compounds are consistent with those reported in the literature, as follows:
[0038] Split ring mangiferin A, colorless needle crystal, NMR data are as follows:.
[0039] 1H-NMR (DMSO-d6, 400MHz): 7.16 (1H, d, J=2.0Hz, 2'-H), 7.06 (1H, dd, J=2.0, 8.0Hz, 5'-H), 6.74 (1H , d, J=8.0Hz, 6′-H), 5.95 (1H, s, 5-H), 4.61 (1H, d, J=9.6Hz, 1″-H), 3.63 (1H, br.d, ca.12Hz, 6″-H), 3.55(1H, dd, J=9.6, 9.6Hz, 2″-H), 3.51(1H, dd, J=3.2, 11.6Hz, 6″-H), 3.22( 1H, m, overlapped, 3″-H), 3.22 (1H, m, overlapped, 4″-H), 3.22 (1H, m, overlapped, 6″-H, 5″-H).
[0040] 13C-NMR (DMSO-d6, 100MHz): 60.3 (6″-C), 69.4 (4″-C), 71.9 (2″-C), 74.6 (1″-C...
Embodiment 3
[0048] Antioxidant Experimental Research on Compounds of the Invention
[0049] Superoxide dismutase-like free radical elimination assay
[0050] Sample group: dissolve the sample in 2.5% dimethyl sulfoxide aqueous solution, add the reaction solution, the reaction solution is 40mM carbonic acid buffer solution (pH 10.2), which contains 100uM xanthine, 100uM ethylenediaminetetraacetic acid dipotassium salt, 25uM NBT (nitroblue tetrazolium), 0.005% fetal bovine serum, about 1.8mU / mL xanthine oxidase, adjust the total volume to 3ml, react in a 37 degree water bath for 20 minutes, add 0.1mL 6mM copper chloride solution to make the reaction Stop, measure the ultraviolet absorption of each sample at a wavelength of 560nm, and calculate the ability to eliminate free radicals according to the following method.
[0051] Wherein, the samples are sample 1-ethanol crude extract; sample 2-component 7-2; sample 3-semangiferin A; sample 4-mangiferin B; sample 5-tetramethylcyclohexyl olefin...
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