免疫亲和柱结合液相色谱串联质谱法测定蜂花粉中黄曲霉毒素

Determination of aflatoxins in bee pollen by immunoaffinity column cleanup coupled with LC-MS/MS

建立了一种测定蜂花粉中黄曲霉毒素（B1、B2、G1、G2）的液相色谱串联质谱方法。样品经V（乙腈）∶V（水）=60∶40提取,通过低温脂肪沉淀,免疫亲和柱净化,之后用甲醇洗脱4种毒素,氮吹后复溶,采用液相色谱-串联质谱（LC-MS/MS）定量分析。方法在3个浓度添加水平的回收率为74.3%~96.5%,精密度低于10.0%。相关系数（r2）均大于0.997。B1、B2、G1、G2的定量限分别为0.05,0.1,0.013,0.025μg/kg。测定30个实际样品,未检出4种毒素。

A simple and sensitive method has been develop for simultaneous determination of aflatoxins （AFB1, AFB2,AFG1, and AFG2） in bee pollen. The sample was extracted with a mixture of acetonitrile/water （60： 40, v/ v） ; fat was precipitated in low temperature, followed by immune affinity column cleanup. The aflatoxins were quanti- fied using liquid chromatography coupled with tandem mass spectrometry （LC-MS/MS）with an eleetrospray ionization （ESI）, and a triple quadrupole （QqQ） analyzer. Recoveries at three levels were in the range of 74.3% -96.5% with RSD less than 10.0%. The correlation coefficients （r2） of the four aflatoxins were higher than 0. 997. The LOQ of B1 ,B2,G1 and G2 were 0.05,0. 1,0. 013,0. 025 g/kg, respectively. The method was simple, accurate and sensi- tive. It was suitable for quantification and confirmation of aflatoxins in bee pollen samples.