HPLC法同时测定双花百合片中绿原酸、表小檗碱、巴马汀、小檗碱的含量

Simultaneous determination of chlorogenic acid,epiberberine,palmatine,berberine in Shuanghua Baihe Tablet by HPLC

目的:建立同时测定双花百合片中绿原酸、表小檗碱、巴马汀、小檗碱4种主要成分的高效液相色谱方法。方法:通过高效液相色谱分析双花百合片甲醇提取液,采用Kromasil C18色谱柱(4.6mm×250mm,5μm),流动相乙腈(A)-0.05mol/L磷酸二氢钾水溶液(C,pH=3)(0-20 min,5%A-20%A;20-65min,20%A),梯度洗脱;体积流量0.8mL/min;检测波长为320nm;柱温30℃;进样量10μL。结果:绿原酸、表小檗碱、巴马汀、小檗碱进样量分别在5.0-500.0μg/mL(r=0.9994)、10.0-500.0μg/mL(r=0.9967)、10.0-1 000.0μg/mL(r=0.9985)、20.0-1 000.0μg/mL(r=0.9994)范围内线性关系良好,平均加样回收率分别为99.63%、101.22%、98.30%、98.93%,RSD值分别为0.97%、1.58%、1.48%、1.93%。结论:本方法简便、准确、重复性好,可为双花百合片的质量标准研究提供参考。

Objective:To establish an HPLC method for simultaneously determining four constituents of Shuanghua Baihe Tablet,including chlorogenic acid,epiberberine,palmatine,and berberine.Methods:The determination analysis was carried out on Kromasil C18 column(4.6mm×250mm,5μm).The mobile phase consisted of acetonitrile(A)-0.05 mol/L Potassium dihydrogen phosphate solution(C,pH=3)(0-20 min,5%A-20%A;20-65 min,20%A) in step gradient elution mode.The flow rate was 0.8mL/min,the detection wavelength was at 320 nm,and the column temperature was 30oC with 10μL injection volume.Results:The experimental data displayed a good linearity in the range of 5.0-500.0μg/mL(r=0.9994) for chlorogenic acid,10.0-500.0μg/mL(r=0.9967) for epiberberine,10.0-1 000.0μg/mL(r=0.9985) for palmatine,and 20.0-1000.0μg/mL(r=0.9994) for berberine,respectively.The average recoveries were 99.63%,101.22%,98.30% and 98.93%,with RSD values of 0.97%,1.58%,1.48% and 1.93%,respectively.Conclusion:This method with strong specificity is simple,accurate and reproducible,and it could provide a reference for the quality standard of Shuanghua Baihe Tablet.