Huang Xiaojing, Wang Shaomin, Mao Dan, Miao Shui, Hu Qing, Ji Shen
Fudan University, School of Pharmacy, 826 Zhangheng Rd, Pudong New District, Shanghai 201203, China; Shanghai Institute for Food and Drug Control, 1500 Zhangheng Rd, Pudong New District, Shanghai 201203, China.
Shanghai Institute for Food and Drug Control, 1500 Zhangheng Rd, Pudong New District, Shanghai 201203, China.
J AOAC Int. 2018 May 1;101(3):633-642. doi: 10.5740/jaoacint.17-0365. Epub 2017 Oct 26.
In our study, a reliable and rapid analytical method for the simultaneous determination of 15 mycotoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, alternariol, agroclavine, citrinin, diacetoxyscirpenol, deoxynivalenol, fumonisin B1, fumonisin B2, ochratoxin A, sterigmatocystin, T-2 toxin, and zearalenone) in liquorice using ultra-HPLC coupled to tandem MS was developed and validated. Due to the complex ingredients in liquorice, we chose a QuEChERS-based extraction procedure as the sample pretreatment. Meanwhile, for the first time, acetate buffer was used to replaced water, which can greatly reduce the concentration of formic acid in acetonitrile, which further reduces the extraction efficiency of impurities. The optimal combination of adsorbents is 150 mg primary secondary amine, 150 mg silica gel, 600 mg octadecylsilane, and 900 mg anhydrous magnesium sulfate. Electrospray ionization in both positive- and negative-ionization modes was applied to detect all the mycotoxins in a single run time of 15 min, with LOQs in the range of 0.125-2.5 μg/kg. The recoveries of determination obtained were in the range of 81.0-104.7%, whereas the analytes could be accurately quantified in the 0.25-625 μg/kg concentration range, with all coefficients being >0.992. Intra- and interday reproducibility were lower than 5.5 and 8.9%, respectively, for all analytical mycotoxins. The validated method was finally applied to screen mycotoxins in 31 batches of real samples collected from drugstores and hospitals in Shanghai, China. Our survey findings show that six mycotoxins were detected, including alternariol, citrinin, deoxynivalenol, fumonisin B1, ochratoxin, and zearalenone, and that the positive rate of mycotoxins was 54.8% in real samples, ranging from 3.37 to 520.6 μg/kg.
在我们的研究中，开发并验证了一种可靠且快速的分析方法，该方法采用超高效液相色谱-串联质谱联用技术同时测定甘草中的15种霉菌毒素（黄曲霉毒素B1、黄曲霉毒素B2、黄曲霉毒素G1、黄曲霉毒素G2、交链孢酚、农吉利麦角碱、桔霉素、二醋酸藨草镰刀菌烯醇、脱氧雪腐镰刀菌烯醇、伏马菌素B1、伏马菌素B2、赭曲霉毒素A、杂色曲霉素、T-2毒素和玉米赤霉烯酮）。由于甘草成分复杂，我们选择基于QuEChERS的提取程序作为样品预处理方法。同时，首次使用醋酸盐缓冲液代替水，这可以大大降低乙腈中甲酸的浓度，进而降低杂质的提取效率。吸附剂的最佳组合为150 mg伯仲胺、150 mg硅胶、600 mg十八烷基硅烷和900 mg无水硫酸镁。采用正离子和负离子电离模式的电喷雾电离，在15分钟的单次运行时间内检测所有霉菌毒素，定量限在0.125 - 2.5 μg/kg范围内。测定回收率在81.0 - 104.7%之间，而分析物在0.25 - 625 μg/kg浓度范围内可准确定量，所有系数均>0.992。所有分析霉菌毒素的日内和日间重现性分别低于5.5%和8.9%。最终将经过验证的方法应用于对从中国上海的药店和医院收集的31批实际样品中的霉菌毒素进行筛查。我们的调查结果表明，检测到六种霉菌毒素，包括交链孢酚、桔霉素、脱氧雪腐镰刀菌烯醇伏马菌素B1、赭曲霉毒素和玉米赤霉烯酮，实际样品中霉菌毒素的阳性率为54.8%，范围为3.37至520.6 μg/kg。
