Mycotoxins are secondary metabolites produced by filamentous fungi. Primary route of human exposure to mycotoxins is the intake of the contaminated food. Minimizing mycotoxin exposure is important for population health, as their chronic toxic effects have been associated with kidney and liver diseases, some types of cancer and immunosuppression. The objective of this work was to develop and validate a multi-class mycotoxin method suitable for exposure monitoring of mycotoxins in human plasma. A sensitive liquid chromatography – mass spectrometry method was developed for 17 mycotoxins: nivalenol (NIV), deoxynivalenol, fusarenon X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, T-2 toxin, HT-2 toxin, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, zearalenone, α-zearalenol (α-ZOL), β-zearalenol, zearalanone, α-zeranol and, and β-zeranol. The method relies on three-step liquid–liquid extraction with ethyl acetate to eliminate the need for immunoaffinity extraction and minimize ionization matrix effects. Chromatographic separation of mycotoxins, including all isomers, was achieved with pentafluorophenyl column and water/methanol mobile phase. Mycotoxin detection and quantitation were performed using high-resolution mass spectrometry on LTQ Velos Orbitrap, in both positive and negative electrospray ionization (ESI(+) and (ESI(−)). The use of 0.02% acetic acid as mobile phase additive for ESI(−) resulted in significant increase in ionization efficiency ranging from 1.7 to 26 times for mycotoxins that ionize better in ESI(−). The optimized method was validated according to FDA guidance procedures. LOQs of all mycotoxins ranged from 0.1 to 0.5 ng/ml, except NIV which resulted in LOQ of 3 ng/ml because of low extraction recovery of this highly polar mycotoxin. Mean intra-day accuracy ranged from 85.8% to 116.4%, and intra-day precision (n = 6) ranged from 1.6% to 12.5% RSD for all mycotoxins except α-ZOL where mean accuracy ranged from 72.9% to 97.2%. Inter-day accuracy and precision were 85.6%–111.5% and 2.7–15.6% RSD respectively, showing good analytical performance of the method for biomonitoring.

霉菌毒素是丝状真菌产生的次级代谢产物。人体接触霉菌毒素的主要途径是摄入被污染的食物。尽量减少霉菌毒素的暴露对于人群健康很重要，因为它们的慢性毒性作用与肾脏和肝脏疾病，某些类型的癌症和免疫抑制有关。这项工作的目的是开发和验证一种适用于人类血浆中真菌毒素暴露监测的多类霉菌毒素方法。开发了一种灵敏的液相色谱-质谱方法，用于17种霉菌毒素：香叶烯酚（NIV），脱氧雪茄烯醇，富沙隆X，3-乙酰基脱氧雪茄烯醇，15-乙酰基脱氧雪茄烯醇，T-2毒素，HT-2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，玉米赤霉烯酮，α-玉米赤霉烯醇（α-ZOL），β-玉米赤霉烯醇，玉米赤霉烯酮，α-玉米醇和β-玉米醇。该方法依赖于乙酸乙酯的三步液-液萃取，从而消除了对免疫亲和萃取的需求，并使电离基质效应最小化。使用五氟苯基色谱柱和水/甲醇流动相实现了霉菌毒素（包括所有异构体）的色谱分离。使用高分辨率质谱法对LTQ Velos Orbitrap进行正向和负向电喷雾电离（ESI（+）和（ESI（-））的真菌毒素检测和定量，使用0.02％乙酸作为ESI的流动相添加剂（-）使在ESI（-）中离子化更好的真菌毒素的电离效率显着提高了1.7到26倍，根据FDA指导程序验证了优化方法，所有真菌毒素的LOQ范围从0.1至0.5 ng / ml ，除了NIV外，由于这种高极性真菌毒素的提取回收率低，导致LOQ为3 ng / ml。除α-ZOL以外，所有霉菌毒素的日内平均准确度范围为85.8％至116.4％，日内准确度（n = 6）范围为1.6％至12.5％RSD。α-ZOL的平均准确度范围为72.9％至97.2％。日间准确性和精密度分别为RSD的85.6％–111.5％和2.7–15.6％，显示了该生物监测方法的良好分析性能。