葡萄籽原花青素的分离纯化及其自由基淬灭活性的研究(上)

Study on Isolation, Purification and the Free-radical Scavenging Effect of Proanthocyanidins from Grape Seed (Ι)

葡萄籽经有机溶剂梯度法及酶法提取，大孔树脂初步纯化，薄层色谱法分离纯化得到原花青素单体和二聚体。结果表明，以60％醇于50℃浸提3次、每次30min（固液比1：7，w／v）效果最好；梯度提取各细分原花青素得率为：（7．73-281．49）mg／100g葡萄籽，其中以乙酸乙酯组分最高，其次为丙酮、乙醇、乙醚。在浓度为0．004g／mL时，淬灭自由基活性最高的是乙醇组分，抑制率为85．13％，其次是乙醚（77．60％）和乙酸乙酯组分（48．64％）。大孔树脂纯化以40％乙醇最佳。以制备薄层法（展开体系：v甲苯：v丙酮：v冰乙酸=2：2：1）分别对乙酸乙酯、乙醚组分进行纯化，前者原花青素单体和二聚体的百分含量分别为33．25％和16．23％，后者为60．64％和10．13％．

The monomer and dimer of pmanthocyanidins in grape seeds were obtained by gradient and enzymatic extraction, isolation with macroporous adsorption resin, and purification by TLC. The results showed that the optimum extraction conditions were as follows： 60% ethanol （1 ：7 w/v）, at 50℃ and 3 times （30 min/time）. The extraction ratios of pmanthocyanidins in gradient extractions were （7.7341-281.494） mg/100g grape seeds and highest content of proanthocyanidins was achieved using the ethyl acetate as extraction reagent followed by acetone, ethanol and ether. The free-radical scavenging effect of the ethanol extracts was higher （with the inhibitory ratio being of 85.13%） than those of ether extracts （77.60%） and ethyl acetate extracts （48.64%）（conc.0.004g/mL）. High content of oligometic pmanthocyanidins isolated by macroporous adsorption resinwere was achived using 40% ethanol. After purification with TLC （ elution system： toluene/acetone/acetic acid=2/2/1）, the yields of monomer and dimmer in ethyl acetate and ether components were 33.25%, 60.64% and 16.23%, 10.13%, respectively.