免疫磁性微球预处理结合超高效液相色谱-串联质谱法快速检测花生和花生油中黄曲霉毒素B_(1)

Rapid determination of aflatoxin B_(1) in peanut and peanut oil by an immunomagnetic beads purification sample pretreatment method combined with ultra performance liquid chromatography-tandem mass spectrometry

目的设计并合成一种功能化免疫磁性微球,结合超高效液相色谱-串联质谱法(ultraperformance liquidchromatography-tandemmassspectrometry,UPLC-MS/MS)快速提取和检测花生和花生油中黄曲霉毒素B1(aflatoxin B1,AFB1)。方法样品经过甲醇-水溶液(70:30,V:V)提取,1.6 mg免疫磁性微球富集净化,用1.00 mL甲醇洗脱,采用Waters ACQUITY UPLC BEH C18色谱柱(50 mm×2.1 mm,1.7μm)分离,电喷雾离子源在正离子模式下分析AFB1的含量。结果该材料能够在15 min实现AFB1的快速提取净化。与UPLC-MS/MS联用,本方法的线性范围在0.1~50.0μg/L,检出限为0.04μg/kg,定量限为0.14μg/kg,相对标准偏差(relativestandard deviation,RSD)为2.2%。将本研究建立的方法用于检测花生及花生油中AFB1,方法回收率大于80%,重复性数据满足实验要求。与基于免疫亲和柱的国家标准方法进行了性能比较,结果与国家标准方法基本一致。结论该方法操作简单、用时短,可用于花生和花生油中AFB1的检测。

Objective To design and synthesize a functional immunomagnetic beads,and combine with ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)to quickly extract and detect aflatoxin B_(1)(AFB_(1))in peanut and peanut oil.Methods The samples were extracted with methanol-water solution(70:30,V:V),enriched and purified with 1.6 mg immunomagnetic beads,and eluted with 1.00 mL methanol,the samples were separated on a Waters ACQUITY UPLC BEH C18 column(50 mm×2.1 mm,1.7μm),and the content of AFB_(1) was analyzed by an electrospray ion source in positive ion mode.Results The material enabled rapid extraction and purification of AFB_(1) in 15 min.When combined with UPLC-MS/MS,the linearity was in the range of 0.1-50.0μg/L,the limit of detection was 0.04μg/kg,the limit of quantification was 0.14μg/kg,and the relative standard deviation(RSD)was 2.2%.The method established in this study was applied for the determination of AFB_(1) in peanut and peanut oil with the recovery rates of over 80%and repeatable data meet experimental requirements.The performance of the proposed method was compared with that of the national standard method based on immunoaffinity column,and the results were basically consistent with that of the national standard method.Conclusion This method is simple and can be used to detect AFB_(1) in peanut and peanut oil.