高效液相色谱-串联质谱法快速测定海鳗鱼肉中3种太平洋雪卡毒素

摘要

目的：建立了一种平行振荡方法萃取海鳗鱼肉中的3种太平洋雪卡毒素(P-CTX-1、P-CTX-2、P-CTX-3),并采用高效液相色谱-电喷雾串联质谱法同时测定P-CTX-1、P-CTX-2、P-CTX-3残留量的分析方法。方法海鳗等样品经甲醇并采用平行振荡方法提取, C18固相萃取柱净化后,采用高效液相色谱-电喷雾串联质谱分析测定。结果以基质标准曲线外标法定量,线性范围为0.01∼50.0 ng/g,相关系数均大于0.999(r≥0.999, n=3), P-CTX-1、P-CTX-2、P-CTX-3的定量限分别为(LOQs,S/N=10)0.009、0.019、0.017 ng/g。以0.05、0.10、0.50 ng/g 3个添加浓度水平进行方法验证,回收率分别为72.9~81.0%、63.0~77.9%、46.3~69.5%,相对标准偏差分别为2.3~2.7%、2.1~12.2%、6.1~15.3%。结论该方法简便快速、灵敏度高,适用于鱼肉中同时快速检测3种太平洋雪卡毒素。%Objective A parallel shaking method was developed to extract three Pacific ciguatoxins (P-CTX-1, P-CTX-2, P-CTX-3) in muscle samples of moray eel and a rapid simultaneous quantification me-thod was optimized to quantify P-CTXs in fish muscle using high performance liquid chromatography coupled with electrospray-tandem mass spectrometry (HPLC-MS/MS).MethodsMuscle samples of moray eel were extracted in methanol using parallel shaker. Crude extract was then cleaned-up by C18 solid-phase extraction (SPE) cartridges. High performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was employed to identify and quantify P-CTXs.ResultsThe quantification of P-CTX-1, P-CTX-2 and P-CTX-3 were performed using matrix-matched calibrations with correlation coefficients above 0.999 (r≥0.999,n=3). The linearity of the calibration curves ranged from 0.01~50.0 ng/g. The limits of quantification (LOQs,S/N=10) of P-CTX-1, P-CTX-2 and P-CTX-3 were 0.009, 0.019, and 0.017 ng/g. Method validation was carried out by spiking three levels of P-CTXs (i.e. 0.05, 0.10, and 0.50 ng/g) to fish muscle. The recovery of P-CTX-1, P-CTX-2 and P-CTX-3 were 72.9~81.0%, 63.0~77.9%, and 46.3~69.5% with relative standard deviations (RSDs) of 2.3%~2.7%, 2.1%~12.2%, and 6.1%~15.3%, respectively.ConclusionThe method is simple, rapid, and high sensitive, and hence suitable for the rapid simultaneous quantification of 3 P-CTXs in fish muscle samples.