在线免疫亲和净化-液相色谱-串联质谱快速测定中草药及中成药中10种真菌毒素

Determination of 10 Mycotoxins in Chinese Herbs and Chinese Medicines by Liquid Chromatography-Tandem Mass Spectrometry Using On-line Immunoaffinity Purifying

将复合毒素免疫亲和柱的柱填料填充至不锈钢柱(35 mm×4.6 mm),制成可重复使用的在线免疫亲和净化柱。在线净化过程的上样溶剂为磷酸盐缓冲液(PBS,pH 7.4),清洗溶剂为水,转移溶剂为乙腈-水(1∶1,V/V)。采用PBS/甲醇溶液提取样品中真菌毒素,提取液直接进行在线免疫亲和净化-液相色谱-质谱/质谱分析,本方法可同时测定中草药及中成药中10种真菌毒素(伏马毒素B1、伏马毒素B2、T-2毒素、HT-2毒素、赭曲霉毒素A(OTA)、玉米赤霉烯酮、α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇和β-玉米赤霉烯醇)。本方法的定量限(LOQ)为1～5μg/kg,各基质在3个水平添加回收率(n=6)为62.3%～107.1%;相对标准偏差(RSD)为2.1%～15.0%。

A reusable on-line immunoaffinity purification column was prepared through packing multitoxin immunoaffinity column filler in stainless column.The loading solution of on-line immunoaffinity purification column was phosphate buffered saline（PBS,pH 7.4）,the washing solution was water,and the transfering solution was（1∶1,V/V） acetonitrile-water.The mycotoxins in Chinese herbs and Chinese medicines were extracted by 75% methanol-PBS.The extract was directly injected into the on-line immunoaffinity column which was coupled with reversed-phase analysis column.After high selective purification of the on-line immunoaffinity column,the mycotoxins were eluted onto analysis column for final separation and analysis.Ten mycotoxins： Toxin T2（T-2）,Toxin HT 2（HT-2）,fumonisin B1,fumonisin B2,ochratoxin A,zearalenone,α-zearalanol,β-zearalanol,α-zearalenol and β-zearalenol,could be detected by the method.The detection limits of the 10 mycotoxins were 1-5 μg/kg.The average recoveries of three levels spiking were 62.3%-107.1%,and the relative standard deviation were ranged from 2.1% to 15.0%.