超高效液相色谱-串联质谱法快速测定建曲中的米酵菌酸

目的： 建立超高效液相-串联质谱法（UPLC-MS/MS法）快速测定中药建曲中生物毒素米酵菌酸的残留。方法： 样品用甲醇超声提取，再用氨水调pH至8，过滤，滤液用Oasis MAX固相萃取柱处理，WATERS HSS T3（100 mm×2.1 mm，1.8 μm）色谱柱分离，以乙腈-含0.1%甲酸的10 mmol·L^-1甲酸铵溶液为流动相梯度洗脱，采用电喷雾（ESI）负离子离子源电离，工作模式为多反应监测模式（MRM）。结果： 米酵菌酸在0.5~100 ng·mL^-1范围内呈良好的线性关系（r>0.999）。稳定性、重复性试验的相对平均偏差（RSD）均小于8.0%。方法回收率为80.3%~85.6%，RSD在3.8%~5.3%之间。方法检测下限（S/N≈3）为2 pg，定量下限（LOQ，S/N≈10）为6 pg。10批样品中7批样品检出米酵菌酸，含量最高达到了115.1 μg·kg^-1，最低的为9.3 μg·kg^-1。结果提示建曲制品存在严重的安全风险。结论： 本法可快速检测建曲中的米酵菌酸残留量，适用于建曲中米酵菌酸的风险监测。本文揭示了建曲中存在生物毒素米酵菌酸污染的现状，为中药的安全性控制提供了研究基础，建议有关部门加强相应的监管工作。

Objective: To establish a ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the determination of bonkrekic acid(a biotoxin)in Jianqu. Methods: UPLC-MS/MS was adopted. The samples were extracted with methanol,then adjust the pH to 8 with ammonia. After filtration the extracts were purified by Oasis Max solid phase extraction columns,seperated by the UPLC column WATERS HSS T3(100 mm×2.1 mm,1.8 μm),and eluted in gradient with acetonitrile-10 mmol·L^-1 ammonium formate solution(contained 0.1% formic acid). Ionization mode was electrospray ionization(ESI)with negative mode and multiple-reaction monitoring mode(MRM)were used. Results: A good linear range of bonkrekic acid as 0.5 ng·ml^-1-100 ng·ml^-1(r>0.999)was found. The relative standard deviations(RSDs)of stability and reproducibility tests were less than 8.0%. The recovery rate of bongkrekic acid were 80.3%-85.6%,and RSDs were between 3.8% and 5.3%. The lower detection limit(S/N ≈ 3)was 2 pg,and the limit of quantification(LOQ)was 6 pg. In 7 out of 10 batches of samples,bonkrekic acid was detected with a range of 9.3-115.1 μg·kg^-1. These results suggested that there are serious safety risks in the Jianqu products. Conclusion: This method can be used to detect residual of bonkrekic aicd in Jianqu. It is applicable to the risk monitoring of bonkrekic acid in Jianqu. This article revealed the status quo of biotoxin bonkrekic acid contamination in Jianqu,which provides a research basis for the safety control of traditional Chinese medicine. It is recommended that the relevant departments should strengthen the corresponding supervision.

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