SPE-HPLC法测定消肿止痛酊中马兜铃酸A的含量

目的:建立消肿止痛酊中马兜铃酸A的限量检查法。方法:通过萃取、过固相萃取小柱对制剂中马兜铃酸A进行提取纯化,应用高效液相色谱法对其进行检测。色谱条件:采用HyperClone ODS 120A C₁₈(4.6 mm×250 mm,5 μm)色谱柱,流动相为0.3%碳酸铵溶液(用20%盐酸溶液调pH至7.5)-乙腈(80:20),流速1.0 mL·min^-1,检测波长250 nm。结果:马兜铃酸A进样量在0.2076~20.76 ng范围内线性关系良好(r=1.000);检测限(S/N=3)为0.21 ng·mL^-1,定量限(S/N=10)为0.52 ng·mL^-1;平均回收率(n=9)为90.1%。结论:所建立的方法简便快捷,重复性符合要求,结果准确可靠,可作为控制消肿止痛酊安全性的检测方法。

Objective:To establish a method for determination the content limit of aristolochic acid A in Xiaozhongzhitong tincture. Methods:Aristolochic acid A was determined from samples and cleaned up by extraction and SPE.The aristolochic acid A was separated on HyperClone ODS 120A C₁₈(4.6 mm×250 mm,5 μm) column with 0.3% ammonium carbonate solution(pH was adjusted to 7.5 with 20% hydrochloric acid)-acetonitrile(80:20) as a mobile phase.The flow rate was 1.0 mL·min^-1 and the detection wavelength was set up at 250 nm.The external standard method was used for quantification. Results:Aristolochic acid A had good linear relationship in the range of 0.2076-20.76 ng and the correlation coefficient was 1.000.The detection limit and quantitation limit of aristolochic acid A in Xiaozhongzhitong tincture were 0.21 and 0.52 ng·mL^-1,respectively.The average recovery(n=9) was 90.1%. Conclusions:The developed method is simple,reproducible,accurate,and can be used as a inspection method for safe use of Xiaozhongzhitong tincture.

1 ChP(中国药典).2010.Vol Ⅰ(一部):1030
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