HPLC法同时测定戊己方提取物中5个成分的含量

目的: 建立高效液相色谱法同时测定戊己方提取物中芍药苷、盐酸巴马汀、盐酸小檗碱、吴茱萸碱、吴茱萸次碱的含量。 方法: 采用Gemini C₁₈ 110A色谱柱(4.6 mm×250 mm,5 μm),流动相A为0.05 mol·L^-1磷酸二氢钾溶液,流动相B为乙腈,梯度洗脱(0~8.5 min,20%B;8.5~9 min,20%B→32%B;9~20 min,32%B;20~21 min,32%B→50%B;21~40 min,50%B),流速1 mL·min^-1,检测波长228 nm,柱温25 ℃。 结果: 在40 min内可完成对戊己方中芍药苷、盐酸巴马汀、盐酸小檗碱、吴茱萸碱、吴茱萸次碱的分离测定,各成分的浓度与峰面积线性关系良好(r=0.9994~0.9996);加样回收率为98.9%~101.1%。 结论: 本方法测定的5个化学成分为戊己方的代表性成分,对其进行含量测定可为含有黄连、吴茱萸、白芍的成方制剂的质量控制提供参考。

Objective: To establish an HPLC method for the simultaneous determination of paeoniflorin,palmatine hydrochloride,berberine hydrochloride,evodiamine and rutaecarpine in Wuji formula. Methods: Simultaneous separation of the five constituents was achieved on Gimini C₁₈ 110A column (4.6 mm×250 mm,5 μm).The mobile phase A was 0.05 mmol·L^-1 monopotassium phosphate and the mobile phase B was acetonitrile.The ratios of the mobile phase B were 20% at 0-8.5 min,20%→32% at 8.5-9 min,32% at 9-20 min,32%→50% at 20-21 min,50% at 21-40 min.The flow rate was 1.0 mL·min^-1.The detection wavelength was set at 228 nm and the column temperature was 25 ℃. Results: Under the condition of gradient elution,the above-mentioned five constituents were separated within 40 minutes.All calibration curves showed good linear regression within test ranges (r=0.9994-0.9996).The recoveries ranged from 98.9% to 101.1%. Conclusion: The five components determined by this method are representative,which can be used for the simultaneous determination of the five constituents in Wuji formula,and can also be taken as reference for quality control of Chinese medicine preparations,including Rhizoma Coptidis,Fructus Evodiae and Paeoniae Radix Alba.