膜荚黄芪毛状根中异黄酮成分的反相高效液相色谱分析

对膜荚黄芪毛状根中6种异黄酮成分进行了反相高效液相色谱法测定。色谱柱为NucleosilC₁₈柱,流动相为甲醇—水(3∶2和3∶1,v/v),检测波长为254nm和280nm。6种异黄酮成分为:10-羟基-3,9-二甲氧基紫檀烷,(3R)8,2’-二羟基-7,4’-二甲氧基异黄烷,芒柄花素(7-羟基-4’-甲氧基异黄酮),8,3’-二羟基-7,4’-二甲氧基异黄酮,2’-羟基-3’,4’-二甲氧基异黄烷-7-O-葡萄吡喃糖甙,毛蕊异黄酮(7,3’-二羟基-4’-甲氧基异黄酮)。异黄酮浓度在2.5～12.5μg范围内与峰面积有良好的线性关系;加样回收率为96.47%～103.33%;精密度试验相对标准偏差为2.57%～6.52%;测得黄芪毛状根中6种异黄酮成分的含量在0.0005%～0.0065%之间。

DETERMINATION OF 6 ISOFLAVONOIDS IN THE HAIRY ROOT CULTURES OF ASTRAGALUS MEMBRANACEUS BY HPLC

An accurate RP-HPLC method for the determination of 10-hydroxy-3, 9-dimethoxypeterocarpan, (3R) 8,2'-dihydroxy-7,4'-dimethoxyisoflavan, formonenetin (7-hydroxy-4'-methoxyisoflavone), 8, 3'-dihydroxy-7, 4'-dimethoxy-isoflavone, 2'-hydroxy-3', 4'-dimethoxyisoflavan-7-O-glucopyranoside and alycosin (7,3'-dihydroxy-4'-methoxyisoflavone) in the hairy root cultures of Astragalus membranaceus was developed. Among them, 10-hydroxy-3, 9-dimethoxypeterocarpan, (3R) 8,2'-dihydroxy-7,4'-dimethoxyisoflavan and 8,3'-dihydroxy-7,4'-dimethoxyisoflavone were new compounds in Astragalus membranaceus. The analytical column was Nucleosil C18. The mobile phases were methanol-water = 3:2 and 1:1 (v/v). The detection wavelengths were 254 nm and 280 nm. Calibration graphs were rectilinear between 2.5 micrograms and 12.5 micrograms. The recoveries were between 96.47% and 103.33%. The relative standard deviations (RSDs) of measurement precision test were 2.57%-6.52%. The flavonoid contents in the hairy root cultures were between 0.0005% and 0.0065% (dry weight).