HPLC测定复方小活络丸中芍药苷含量和乌头碱的限量

Determination of Paeoniflorin and Limit Examination of Aconitine in Compound Xiaohuoluo Pills By HPLC

摘要:

目的: 建立HPLC测定复方小活络丸中芍药苷含量的方法以及乌头碱的限量检查。 方法: 采用 HPLC,色谱柱为十八烷基硅烷键合硅胶(4.6 mm×150 mm,5 μm);芍药苷检测以乙腈-0.1%磷酸溶液(14:86)为流动相,柱温 30℃,流速1.0 mL·min^-1,检测波长 230 nm。乌头碱检测乙腈-四氢呋喃(25:15)为流动相A,以0.1 mol·L^-1醋酸铵溶液(每1 000 mL 加冰醋酸0.5 mL)为流动相B,梯度洗脱,柱温 35℃,流速0.8 mL·min^-1,检测波长 235 nm。 结果: 芍药苷在0.035 5~0.709 6 μg 线性关系良好(r=0.999 9),平均加样回收率101.9%(RSD 1.86%,n=6);乌头碱在0.066 0~0.495 0 μg线性关系良好(r=0.999 8),平均加样回收率102.0%(RSD 1.75%,n=6)。 结论: 方法简便,准确,重复性好,可用于复方小活络丸芍药苷的含量测定和乌头碱的限量检查。

Abstract:

Objective: To develop a HPLC method for determination of paeoniflorin and limit examination of aconitine in compound Xiaohuoluo pills. Method: For paeoniflorin, C₁₈ column (4.6 mm×150 mm, 5 μm) was used at 30℃; with the mobile phase was acetonitrile-0.4% phosphate solution (14: 86). The detection wavelength was 230 nm and the flow rate wsa set at 1.0 mL·min^-1. For aconitine, C₁₈ column (4.6 mm×150 mm, 5 μm) was used at 35℃; with the mobile phase was acetonitrile-tetrahydrofuran(25: 15)and 0.1 mol·L^-1 ammonium acetate solution by gradient elution. The detection wavelength was 235 nm and the flow rate wsa set at 0.8 mL·min^-1. Result: The linear range of Paeoniflorin was 0.035 5-0.709 6 μg(r=0.999 9).The average recoveries was 101.9% and RSD was 1.86%(n=6). The linear range of aconitine was 0.066 0-0.495 0 μg(r=0.999 8).The average recoveries was 102.0% and RSD was 1.75%(n=6). Conclusion: The method is simple, accurate, stable and can be used for determination of paeoniflorin and limit examination of aconitine in compound Xiaohuoluo pills.