HPLC-UV法测定黄芪中黄酮类化合物含量

Determination of flavonoids content in Astragali Radix by HPLC-UV

采用HPLC-UV法同时测定黄芪中毛蕊异黄酮苷、毛蕊异黄酮、芒柄花苷以及芒柄花素含量。样品经浸泡-超声处理,流动相为乙腈-0.1%甲酸水溶液,低压梯度洗脱;流速1.0 mL/min;进样量25μL;柱温30℃;检测波长254 nm。结果显示,毛蕊异黄酮苷在线性范围18.70~187.20μg/mL色谱峰峰面积与浓度线性关系良好,相关系数为0.999 7;毛蕊异黄酮在线性范围1.68~16.80μg/mL色谱峰峰面积与浓度线性关系良好,相关系数为0.999 4;芒柄花苷在线性范围7.50~75.00μg/mL色谱峰峰面积与浓度线性关系良好,相关系数为0.999 5;芒柄花素在线性范围4.90~49.20μg/mL色谱峰峰面积与浓度线性关系良好,相关系数为0.999 8。精密度、稳定性、适应性、重复性试验以及加样回收率均符合要求。该方法简便快捷,精密度高,重现性好,稳定性高,可用于黄芪药材中多成分含量的同时测定。

HPLC-UV method was used to determine the content of calycosin glycoside,calycosin,ononin and fermononetin in Astragalus membranaceus.The samples were treated by immersion ultrasonic treatment.The mobile phase was acetonitrile-0.1% formic acid aqueous solution with low pressure gradient elution;the flow rate was 1.0 mL·min^(-1);the injection volume was 25 μL;the column temperature was 30℃;the detection wavelength was 254 nm.The results showed that the linear range of calycosin glycoside,calycosin,ononin and fermononetin was 18.70-187.20 μg·mL^(-1),1.68-16.80 μg·mL^(-1),7.50-75.00 μg·mL^(-1) and 4.90-49.00 μg·mL^(-1) with the correlation coefficient of 0.999 7,0.999 4,0.999 5 and 0.999 8,respectively,indicating that the peak area had a good linear relationship with the concentration.The precision,stability,adaptability,repeatability test and sample recovery were all in line with the requirements.The method is simple,rapid,accurate,reproducible and stable.It can be used for the simultaneous determination of multiple components in Aastragali Radix.