HPLC-VWD-ELSD法同时测定黄甲软肝颗粒中6个成分的含量

Simultaneous determination of six components in Huangjia Ruangan granules by HPLC-VWD-ELSD

目的:采用高效液相色谱-紫外检测器-蒸发光散射检测器(HPLC-VWD-ELSD)法同时测定黄甲软肝颗粒中没食子酸、葛根素、芍药苷、三七皂苷R_(1)、黄芪甲苷和丹参酮Ⅱ_(A)的含量。方法:采用Waters Symmetry C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇(A)-0.1%醋酸水溶液(B),梯度洗脱,流速为1.0 m L·min^(-1)。没食子酸、葛根素和丹参酮Ⅱ_(A)采用VWD进行检测,检测波长为270 nm;芍药苷,三七皂苷R_(1)和黄芪甲苷采用ELSD进行检测,漂移管温度为70℃,载气流速2.0 L·min^(-1)。结果:没食子酸、葛根素、芍药苷、三七皂苷R_(1)、黄芪甲苷和丹参酮Ⅱ_(A)进样量分别在1.444-28.88、2.118-42.36、1.074-21.48、0.2180-4.360、0.1151-2.302和0.06670-1.334μg范围内与峰面积呈良好的线性关系;平均回收率(n=6)分别为93.8%、98.8%、97.5%、103.0%、100.6%和96.6%,RSD分别为1.4%、0.55%、3.3%、4.0%、3.9%和2.3%。5批黄甲软肝颗粒中上述6个成分的含量测定结果分别为2.612-3.203、9.155-10.342、7.044-9.863、0.622-0.841、1.529-1.697和0.719-0.957 mg·g^(-1)。结论:所建立的方法简便、快捷,重复性好,可用于黄甲软肝颗粒中6个成分的含量测定,为黄甲软肝颗粒全面的质量控制提供科学依据。

Objective:To develop a method of high performance liquid chromatography-variable wavelength detector-evaporative light scattering detection(HPLC-VWD-ELSD)for simultaneous determination of gallic acid,puerarin,paeoniflorin,notoginsenoside R_(1),astragaloside and tanshinoneⅡ_(A) in Huangjia Ruangan granules.Methods:HPLC assay was performed on a Waters Symmetry-C_(18) column(250 mm×4.6 mm,5μm)with a mixture of methanol and 0.1%aqueous acetic acid as the mobile phase in gradient elution at the flow rate of 1.0 mL·min^(-1).Detection wavelength for gallic acid,puerarin and tanshinoneⅡ_(A) was set at 270 nm and ELSD was used for detection of paeoniflorin,notoginsenoside R_(1) and astragaloside.The temperature of the drift tube was maintained at 70℃and the nebulizing gas flow rate was 2.0 L·min^(-1).Results:The linear ranges of gallic acid,puerarin,paeoniflorin,notoginsenoside R_(1),astragaloside and tanshinoneⅡ_(A) were 1.444-28.88,2.118-42.36,1.074-21.48,0.2180-4.360,0.1151-2.302 and 0.06670-1.334μg,respectively.The average recoveries(n=6)were 93.8%with RSD of 1.4%,98.8%with RSD of 0.55%,97.5%with RSD of 3.3%,103.0%with RSD of 4.0%,100.6%with RSD of 3.9%,96.6%with RSD of 2.3%,respectively.The contents of the above-mentioned components in 5 samples were 2.612-3.203,9.155-10.342,7.044-9.863,0.622-0.841,1.529-1.697 and 0.719-0.957 mg·g^(-1),respectively.Conclusion:The method is simple,rapid and reproducible.It can be used for the determination of the six components in Huangjia Ruangan granules,providing a scientific basis for the comprehensive quality control of Huangjia Ruangan granules.