中药材中4种马兜铃酸的检测方法研究

Extraction and Determination of Four Different Aristolochic Acids in Aristolochic Herbs and Chinese Patent Drug

目的:建立同时检查AA Ⅰ、AA Ⅱ、AA Ⅲa、AA ⅠVa 4种马兜铃酸的方法。方法:药材经甲醇-水(70∶30,v/v)溶解,超声提取1 h,经离心取上清液后,采用高效液相色谱法测定药材中AA Ⅰ、AAII、AA Ⅲa、AA ⅠVa 4种马兜铃酸的含量。以萘普生为内标物,采用内标法进行定量。结果:在1-100μg·mL-1范围呈良好的线性关系,4种马兜铃酸中的相关系数(r2)均大于0.9990。在马兜铃药材样品基质低、中、高3个加标水平条件下,AA Ⅰ的回收率为97.2%-120.6%,AA Ⅱ的回收率为58.3%-98.0%,AA Ⅲa的回收率为73.6%-86.0%,AA ⅠVa的回收率为82.5%-111.4%,相对标准偏差为0.16%-6.78%。4种马兜铃酸的日内相对标准偏差在0.19%-4.04%之间,日间相对标准偏差在0.23%-5.41%之间。结论:该方法的准确性、灵敏度和精密度均符合药物中马兜铃酸含量的检测要求。

This study aims to explore the content determination and extraction methods of four aristolochic acids （i.e., AA I, AA II, AA Ilia, AA IVa） in herbs. The extraction method is one-hour ultrasonic extraction in methanol-water （70：30, v/v） solution. And the supernatant liquor is analyzed through HPLC. The results showed that there is a good linear relationship within 0.1 - 100 p,g/mL with more than 0.9990 correlation coefficient （r2）. The recovery rate of AA I is between 97.2% -120.6% and that of AA IVa is between 82.5% - 111.4% through the results of 5 - 50 ug/mL fortified concentration. The recovery rate of AA II is between 58.3% - 98.0% and that of AA IIIa is between 77.5% - 86.0% through the results of 2 -20 ug/mL fortified concentration. The intra-day relative standard deviation （RSD, n = 5） is between 0.19% - 4.04% and the inter-day RSD （n = 5） is between 0.23% - 5.41%. The accuracy, sensitivity and precision can meet the requirements of content determination of aristolochic acids in medicine. Therefore, it is appropriate to be used in quantitative analysis and determination of medicine extracting solution which contains aristolochic acids.