秀丽莓中鞣花酸的含量测定和抗氧化活性评测及对接研究

摘要

Objective To establish a method to determine the content and to estimate the antioxidant activity of ellagic acid in Rubus amabilis .Methods The RP-HPLC separation was performed on a SunFireTM C18 analytical column (150 mm × 4 .6 mm ,5μm) .The mobile phase was acetonitrile-water containing 4 mL · L -1 H3 PO4 (20:80)with a flow rate of 1 .0 mL · min-1 .The detection wavelength was set at 254 nm ,and the column temperature was 35 ℃ .The computer molecular simulation and 2 ,2′-diphenyl-1-pic-rylhydrazyl (DPPH · ) antioxidation in vitro methods were used together to estimate the antioxidant activity of ellagic acid . Results The liner range of ellagic acid was 0 .08192-0 .40960 μg (r=0 .9997 ,n=6) .The average recovery (n=6) was 99 .51％(RSD=2 .00％ ) .Ellagic acid was functioned at the-NH2 groups in receptor proteins by the hydrogen bonds between -OH groups andπ-π bonds among benzene rings ,which resulted in the more significant antioxidant activity with vitamin C .Conclusion The method is simple ,stable and accurate with a good reproducibility ,and can be used as a method for quality control and evaluation of the main antioxidation component ,ellagic acid ,in Tibetan medicine Rubus amabilis .The method can be used in the quality evalua-tion of the further development of the plant .%目的 建立秀丽莓中鞣花酸的含量测定方法并对其抗氧化活性进行研究.方法 采用RP-HPLC法,SunFireTM C18色谱柱(150 mm×4.6 mm,5μm),乙腈-4 mL·L-1磷酸水(20:80)为流动相;流速:1.0 mL·min-1;检测波长:254 nm;柱温:35℃.有机结合计算机分子模拟及体外DPPH·抗氧化活性评价方法对其可能的作用机制及活性进行深入探讨.结果 鞣花酸的线性范围为0.08192～0.40960μg(r=0.9997,n=6);平均回收率(n=6)为99.51％(RSD=2.00％).该成分可通过结构中的酚羟基经氢键、苯环经π-π键作用与受体蛋白的氨基结合起效,具有较维生素C更强的抗氧化活性.结论 该方法可迅速、准确地测定秀丽莓中的鞣花酸含量,可作为该药材中主要抗氧化成分鞣花酸质量控制和评价的有效方法,为该药材的进一步开发提供判断方法.