液相色谱-串联质谱法测定饲料中黄曲霉毒素的研究

Determination of Aflatoxins in Feeds by Liquid Chromatography-Tandem Mass Spectrometry

  • 摘要: 建立了液相色谱串联质谱法测定饲料中6种黄曲霉毒素(包括黄曲霉毒素B1、B2、G1、G2、M1、M2)的方法。试样中的黄曲霉毒素经84%乙腈溶液超声提取后,用正己烷脱脂,过霉菌毒素多功能固相萃取柱净化,氮气吹至干,用流动相溶解后进行液相色谱串联质谱法测定。方法的最低检出限为0.2 μg•kg-1,定量限均为0.5 μg•kg-1;标准工作液在0.5~100 μg•L-1的范围内线性良好;加标浓度在1.0~100 μg•kg-1范围内,饲料中黄曲霉毒素的回收率在66.1%~108%之间,批内相对标准偏差≤17.8%,能满足相关法规要求。

     

    Abstract: Totally 6 Aflatoxins in feeds were determined by high performance liquid chromatography combined with electrospray ionization triple quadrupole tandem mass spectrometry(HPLC-MS/MS) under the multiple reaction monitoring (MRM) mode, and especially focused on the optimization of extraction, clean-up, HPLC separation and MS/MS parameters of analytes. The Aflatoxins B1、B2、G1、G2、M1 and M2 were extracted by 84% of acetonitrile aqueous solution, purified by passing through multifunctional cartridges. The elution was evaporated by nitrogen blow and dissolved by mobile phase, and then assayed by HPLC-MS/MS. The limits of detection(LOD) are 0.2 μg•kg-1, and the limits of quantitation(LOQ) are 0.5 μg•kg-1. The calibration curves are linear between 0.5 μg•L-1 and 100 μg•L-1 for Aflatoxins B1、B2、G1、G2、M1 and M2. Recoveries of those mycotoxins in feeds are 66.1%—108%, and inter-relative standard deviation(n=5) is less than 17.8% at spiked level of 1.0—100 μg•kg-1. The method is simple, sensitive and reliable for feed safety.

     

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