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

Simultaneous determination of geniposide,paeoniflorin,baicalin,berberine hydrochloride and emodin in Huanglian Shuangqing pills by HPLC

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,5μm),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;30～35min,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0.08～0.4μg(r=0.9998)、0.128～0.768μg(r=0.9996)、0.128～0.896μg(r=0.9996)、0.10～0.554μg(r=0.9997)和0.08～0.48μg(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, ba- iealin, berberine hydrochloride and emodin in Huanglian Shuangqing pills. Methods：The analysis was performed on a Zorbax Eclipse XDB - CI8 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 % →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 hydrochlo- ride 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 ba- icalin,0. 10 -0. 554 μg（r =0. 9997） for herberine hydrochloride and 0. 08 -0. 48 μg（r =0. 9996） for emodin,re- spectively. 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, berherine hydrochloride and emodin in Huanglian Shuangqing pills.