高效液相色谱串联质谱法测定花生及制品中的五种真菌霉素

Determination of Five Mycotoxins in Peanuts and Products by HPLC-MS/MS

摘要:

建立了高效液相色谱-串联质谱（HPLC-MS/MS）测定花生及制品中黄曲霉毒素B1、黄曲霉毒素M1、脱氧雪腐镰刀菌烯醇、赭曲霉毒素A、玉米赤霉烯酮五种真菌霉素的快速分析方法。用甲醇-水（55:45，V/V）对样品进行提取，采用真菌毒素免疫亲和柱萃取，在ESI+模式下采用多反应监测(MRM)模式进行检测。目标物在C18色谱柱上实现了有效分离，在6 min内完成一个样品的分析，相关系数(r2，n=6)大于0.999，检测结果稳定、灵敏。黄曲霉毒素B1、黄曲霉毒素M1、脱氧雪腐镰刀菌烯醇线性范围0.5~50.0 μg/L，检出限为0.05 μg/kg (LOD，S/N=3)，赭曲霉毒素A、玉米赤霉烯酮线性范围5.0~100.0 μg/L，检出限为0.5 μg/kg (LOD，S/N=3)，方法回收率为86.8~102.7%，精密度RSD为0.36~4.79%。该方法快速、灵敏，适用于花生及制品中黄曲霉毒素B1、黄曲霉毒素M1、脱氧雪腐镰刀菌烯醇、赭曲霉毒素A、玉米赤霉烯酮五种真菌霉素的检测与确证。

Abstract:

A method based on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for simultaneous determination of five mycotoxins, namely aflatoxin B1 (AFB1) , aflatoxin M1 (AFM1), Deoxynivalenol (DON), Zearalenone (ZEA) and OchratoxinA (OTA), in peanut and its products. The samples were extracted with methanol-aqueous solution (55:45, V/V), cleaned up and enriched by mycotoxin affinity column, and then analyzed by HPLC-MS/MS under the multiple reaction monitoring (MRM) mode. All the analytes were successfully detected in positive ion mode within 6 min, and the correlation coefficients (r2, n=6) were more than 0.999. The linear ranges for AFB1, AFM1, and DON were between 0.5 μg/L to 50.0 μg/L, with detection limit of (LOD, S/N=3) 0.05 μg/kg. While ZEA and OTA were ranging from 5.0 μg/L to 100.0 μg/L, and the detection limit (LOD, S/N=3) was 0.5 μg/kg. The spiked recoveries were in the range of 86.8~102.7%, and relative standard deviations (RSD, n=3) were 0.36~4.79%. The method was simple, effective and sensitive, and was suitable for the determination and confirmation of five mycotoxins in peanuts and products samples.