HPLC法同时测定黄连双清丸中栀子苷、芍药苷、黄芩苷、盐酸小檗碱和大黄素的含量

目的: 建立同时测定黄连双清丸中栀子苷、芍药苷、黄芩苷、盐酸小檗碱和大黄素含量的HPLC法。 方法: 采用Zorbax Eclipse XDB-C₁₈色谱柱(4.6 mm×150 mm,5 μm),流动相为乙腈(A)-0.2%磷酸水溶液(B),采用梯度洗脱(0~5 min,12%A;5~10 min,12%A→20%A;10~20 min,20%A→25%A;20~25 min,25%A→30%A;25~30 min,30%A→65%A;30~35 min,65%A→90%A),流速为1.0 mL·min^-1,柱温为室温,检测波长分别为240 nm(栀子苷、芍药苷),275 nm(黄芩苷、盐酸小檗碱、大黄素)。 结果: 栀子苷、芍药苷、黄芩苷、盐酸小檗碱和大黄素进样量分别在0.08~0.4 μg(r=0.9998)、0.128~0.768 μg(r=0.9996)、0.128~0.896 μg(r=0.9996)、0.10~0.554 μg(r=0.9997)和0.08~0.48 μg(r=0.9996)范围内线性关系良好;平均加样回收率(n=6)分别为101.3%、98.5%、100.2%、97.4%和95.9%,RSD分别为2.8%、1.7%、2.2%、1.3%和1.9%。 结论: 本法适用于同时测定黄连双清丸中栀子苷、芍药苷、黄芩苷、盐酸小檗碱和大黄素的含量。

Objective: To develop an HPLC method for simultaneous determination of gardenoside,paeoniflorin,baicalin,berberine hydrochloride and emodin in Huanglian Shuangqing pills. Methods: The analysis was performed on a Zorbax Eclipse XDB-C₁₈ column(150 mm×4.6 mm,5 μm) with gradient elution(0-5 min,12%A;5-10 min,12%A→20%A;10-20 min,20%A→25%A;20-25 min,25%A→30%A;25-30 min,30%A→65%A;30-35 min,65%A→90%A) by using the mobile phase of acetonitrile(A)-0.2% phosphoric acid solution(B).The flow rate was 1.0 mL·min^-1 and the column temperature was maintained at room temperature,the detection wavelengths were set at 240 nm(for gardenoside and paeoniflorin) and 275 nm(for baicalin,berberine hydrochloride and emodin),respectively. Results: The calibration curves were linear within the range of 0.08-0.4 μg(r=0.9998) for gardenoside,0.128-0.768 μg(r=0.9996) for paeoniflorin,0.128-0.896 μg(r=0.9996) for baicalin,0.10-0.554 μg(r=0.9997) for berberine hydrochloride and 0.08-0.48 μg(r=0.9996) for emodin,respectively.The recoveries(n=6) of gardenoside,paeoniflorin,baicalin,berberine hydrochloride and emodin were 101.3%(RSD=2.8%),98.5%(RSD=1.7%),100.2%(RSD=2.2%),97.4%(RSD=1.3%),and 95.9%(RSD=1.9%),respectively. Conclusion: The method can be used for simultaneous determination of gardenoside,paeoniflorin,baicalin,berberine hydrochloride and emodin in Huanglian Shuangqing pills.