RP-HPLC法测定牛黄上清丸中栀子苷、芍药苷、黄芩苷、小檗碱的含量

目的:建立牛黄上清丸中栀子苷、芍药苷、黄芩苷和小檗碱的含量测定方法。方法:采用高效液相色谱(RP-HPLC)梯度洗脱,在不同波长下测定含量。色谱条件:Zorbax Eclipse XDB-C18键合硅胶(4.6 mm×150 mm,5μm),流动相乙腈为0.2%磷酸水梯度洗脱,流速1.0 m L/min,柱温为20℃,检测波长分别为240 nm(栀子苷、芍药苷),275 nm(黄芩苷),350 nm(盐酸小檗碱)。结果:栀子苷、芍药苷、黄芩苷和盐酸小檗碱分别在0.112~0.84μg(r=0.9998),0.20~1.50μg(r=0.9992),0.208~3.64μg(r=0.9990),0.232~2.32μg(r=0.999 7)范围内线性关系良好,平均加样回收率分别为98.62%,100.30%,99.80%,98.92%,RSD分别为1.66%、1.93%、0.94%、0.95%。结论:该方法操作简便,结果准确,精密度高,重现性良好,可用于同时测定牛黄上清丸(大蜜丸)中栀子苷、芍药苷、黄芩苷和盐酸小檗碱的含量,可为提升、完善牛黄上清丸的质量标准及含量测定的方法提供科学依据。

Determination of Jasminoidin,Peoniflorin, Baicalin and Berberine Hydrochloride in NiuHuang ShangQing Pills by RP-HPLC

Objective: To establish the method for content determination of jasminoidin, peoniflorin, baicalin and berberine hydrochloride inNiuHuangShangQingpills. Methods: RP-HPLC was adopted for gradient elution, and the contents were detected under different wavelength. Chromatographic conditions: Zorbax Eclipse XDB-C18 bonded silica gel chromatographic column(4.6 mm×150 nm, 5μm), mobile phase: acetonitrile: 0.2% phosphoric acid solution, gradient elution, flow rate was 1.0 mL/min, column temperature was 20℃, detection wavelengths were 240 nm (jasminoidin, peoniflorin), 275 nm (baicalin) and 350 nm (berberine hydrochloride). Results: Jasminoidin, peoniflorin, baicalin and berberine hydrochloride presented better linear relationship in the ranges between 0.112 and 0.84μg(r=0.999 8), 0.20 and 1.50μg(r=0.999 2), 0.208 and 3.64μg (r=0.999 0), 0.232 and 2.32μg(r=0.999 7), av-erage recovery rates were 98.53%, 100.30%, 99.80% and 98.62% respectively. RSD were 1.66%, 1.93%, 0.94% and 0.95%. Conclusion: The method, easy to operate, accurate and reproducible, could be used to determine the contents of jasminoidin, peoniflorin, baicalin and berberine hydrochloride inNiuHuangShangQingpills simultaneously, and it could provide scientific evidence for improving quality control and content determination methods ofNiuHuang ShangQing pills.