药食同源中药材中16种真菌毒素的测定与分析

李凤华; 李作华; 杨丽; 员永春; 王素香; 王爱香; 李蔚; 郑凤家

doi:10.13386/j.issn1002-0306.2021080109

药食同源中药材中16种真菌毒素的测定与分析

Determination and Analysis of 16 Mycotoxins in Medicinal and Edible Traditional Chinese Medicine

摘要

摘要: 目的：建立同位素标记-超高效液相色谱-串联质谱法（UPLC-MS/MS）测定药食同源中药材中16种真菌毒素的分析方法，并利用该方法对市售的483份药食同源样品进行检测分析。方法：样品用乙腈-水（50/50，V/V）提取，MycoSpin^TM 400多毒素净化柱净化，经Acquity UPLC BEH C₁₈色谱柱(100 mm×2.1 mm，1.7 μm)分离，质谱采用电喷雾电离源（ESI），多反应监测模式（MRM）进行检测分析，同位素内标法进行定量。结果：16种真菌毒素标准曲线线性关系良好（R>0.998），方法的检出限在0.1~4.0 μg/kg之间，高、中、低3个不同浓度加标回收率为83.4%~102.3%，相对标准偏差（RSD，n=6）为2.08%~13.6%。483份样品中，共检出10种真菌毒素，其余6种真菌毒素均未检出，检出率最高的毒素化合物为玉米赤霉烯酮（ZEN），阳性样品中平均含量为71.2 μg/kg，并且有3.11%样品超过国家食品安全标准规定的参考限量。结论：该方法采用同位素稀释，多毒素净化柱对样品进行净化，降低了药食同源样品中基质干扰，方法准确、快速，满足测定方法的要求，可用于大批量样品中真菌毒素的多残留检测分析。

Abstract: Objective: An analytical method was established for the determination of 16 mycotoxins in traditional Chinese medicines (TCM) by isotope labeling-ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), and 483 medicinal and edible homologous samples from the markets were detected using this method. Method: The samples were extracted with acetonitrile-water (50/50, V/V), and then purified by the MycoSpin^TM 400 multifunction clean-up columns. Then the samples were detected and confirmed by UPLC-MS/MS, and quantified by isotope labeled internal standards. Rapid separation of 16 mycotoxins was successfully achieved on an Acquity UPLC BEH C₁₈ column（100 mm×2.1 mm, 1.7 μm）with gradient elution. Simultaneous acquisition was performed in multiple reaction monitoring (MRM) mode with electrospray ionization (ESI) source operated in both positive and negative ionization modes. Result: The established method provided good linearities for the 16 mycotoxins within their respective linear ranges with high correlation coefficients (R>0.998), and the detection limits of this method were 0.1~4.0 μg/kg. The average recoveries of the 16 mycotoxins ranged from 83.4% to 102.3% at the three spiked levels, and the relative standard deviations (RSD, n=6) were in the range of 2.08%~13.6%. In the detection of 483 actual samples, 10 mycotoxins were detected, and the other 6 mycotoxins were not detected. The toxin compound with the highest detection rate was zearalenone (ZEN), with an average content of positive samples 71.2 μg/kg, and 3.11% of the samples exceeded the reference limit specified in the national food safety standard. Conclusion: This method used isotope dilution and multifunction clean-up columns to purify the samples, which reduced the matrix interference in the medicinal and edible homologous samples. The detection limits could meet the requirement of methods. The method was accurate and rapid, and could be used for the detection and analysis of multi residues of mycotoxins in a large number of samples.