SPE-HPLC法测定消肿止痛酊中马兜铃酸A的含量

Determination of aristolochic acid A in Xiaozhongzhitong tincture by SPE-HPLC

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,5μm)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 Xiaozhong- zhitong 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 Cls （4.6 mm× 250 mm, 5μm） 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 - and the detection wavelength was set up at 250 nm. The exter- nal 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%. Conclusion： The developed method is simple, reproducible, accurate, and can be used as a inspection method for safe use of Xiaozhongzhitong tincture.