HPLC法测定十三味逐瘀合剂中游离大黄酚含量

Determination of free chrysophanol content in Shisanwei Zhuyu Mixture by HPLC

目的建立十三味逐瘀合剂中大黄酚高效液相含量测定的方法。方法采用高效液相色谱法(HPLC法)对十三味逐瘀合剂中游离大黄酚含量进行测定,从提取溶剂、取样量、提取时间等影响因素中选出最优条件,建立完整的含量测定方法。结果用thermos C18色谱柱(250 mm×4.6 mm,5μm);以甲醇-0.1%磷酸(75∶25,V/V)为流动相,采用等度洗脱法,254 nm波长的色谱条件;以甲醇为浸膏提取溶剂,超声30 min的提取方法;大黄酚质量浓度在1.63~16.32μg/mL(R^(2)=0.9995)范围内具有良好的线性关系;平均回收率为104.17%;测得十三味逐瘀合剂中游离大黄酚平均含量为15.32μg/mL。结论本实验所建立的方法操作简单,专属性强,重现性好,可用于十三味逐瘀合剂中游离大黄酚的含量测定。

Objective To establish a method for the determination of high performance liquid phase content of chrysophanol in Shisanwei Zhuyu Mixture.Methods High performance liquid chromatography(HPLC)method was used to quantify the concentration of free chrysophanol in Shisanwei Zhuyu Mixture.The optimal conditions were selected from influencing factors such as extraction solvent,sampling volume,and extraction time to establish complete content determination method.Results Chromatography was performed on a thermos C18 column(250 mm×4.6 mm,5μm)using a mobile phase of methanol-0.1%phosphoric acid(75∶25,V/V)with isocratic elution and a wavelength of 254 nm.The extraction method was performed using methanol as the solvent and ultrasound for 30 minutes.The mass concentration of chrysophanol exhibited good linearity in the range of 1.63-16.32μg/mL(R=0.9995).Average recovery rate was 104.17%,and the average content of free chrysophanol in Shisanwei Zhuyu Mixture was measured to be 15.32μg/mL.Conclusion The method established in this experiment is simple,specific and reproducible,and can be used for the determination of free chrysophanol in Shisanwei Zhuyu Mixture.