免疫亲和柱净化-超高效液相色谱串联质谱法同时测定火锅底料中的5种真菌毒素

Simultaneous Determination of 5 Mycotoxins in Hot Pot Seasoning by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry with Immunoaffinity Column Clean-Up

目的:建立复合免疫亲和柱净化、超高效液相色谱串联质谱检测火锅底料中的黄曲霉毒素B_(1)(Aflatoxins B_(1),AFB_(1))、黄曲霉毒素B_(2)(Aflatoxins B_(2),AFB_(2))、黄曲霉毒素G_(1)(Aflatoxins G_(1),AFG_(1))、黄曲霉毒素G_(2)(Aflatoxins G_(2),AFG_(2))、赭曲霉毒素A(Ochratoxin A,OTA)的方法。方法:样品提取后,经复合免疫亲和柱净化,以0.1%甲酸水和甲醇为流动相梯度洗脱,用Agilent EclipsePlus C_(18)RRHD色谱柱分离,ESI+进行多反应监测,内标法定量。结果:5种真菌毒素的线性范围在0.1~10.0 ng/mL,相关系数(r)>0.999,检出限0.1μg/kg,定量限0.3μg/kg。3个加标水平下(0.2、5.0、10.0μg/kg)的回收率在81.38%~97.87%之间,相对标准偏差为0.79%~6.18%。结论:该方法快速准确,可用于火锅底料中5种真菌毒素的定性、定量检测。

Objective:A method for the determination of aflatoxins B_(1)(AFB_(1)),aflatoxins B_(2)(AFB_(2)),aflatoxins G_(1)(AFG_(1)),aflatoxins G_(2)(AFG_(2)) and ochratoxin A in hot pot seasoning by ultra performance liquid chromatography-tandem mass spectrometry with immunoaffinity column clean-up was established.Methods:After extraction,the samples were purified by immunoaffinity column,eluted with 0.1%formic acid water and methanol as mobile phase gradient,separated by Agilent EclipsePlus C_(18) RRHD chromatographic column,ESI+ was used for multiple reaction monitoring,and internal standard method was used for quantification.Results:Good linearity was obtained for 5 mycotoxins within the concentration range of 0.10–10.0 ng/mL,with the correlation coefficients(r) higher than 0.999,the limit of detection and the limit of quantification of the 5 mycotoxins were 0.1 μg/kg and 0.3 μg/kg.The average recovery rates at three concentrations of 0.20,5.0,10.0 μg/kg was in the range of 81.51%–97.87%,with relative standard deviations of 0.79%–6.18%.Conclusion:The results showed that this method could be used to determine the quality and quantity of 5 mycotoxins rapidly and accurately.