免疫磁珠富集结合酶联免疫吸附法检测酱油中黄曲霉毒素B_1

Immunomagnetic Bead Enrichment and ELISA for Detection of Aflatoxin B_1 in Sauce

将活泼酯法活化后的羧基化超顺磁珠与辛酸一硫酸铵法纯化的抗黄曲霉毒素B，单克隆抗体偶联，获得黄曲霉毒素B，（AFB。）免疫磁珠。比较不同粒径大小的免疫磁珠的富集效率，优化磁珠偶联抗体的条件以及富集AFB。的反应条件，初步建立了以免疫磁珠富集结合酶联免疫吸附法检测酱油基质中的AFB，的方法。结果表明：酶联免疫吸附法在0．05～0．3gg／kg范围内具有良好的线性关系，相关系数为0．9842。在酱油中添加加1-7μg／kgAFB，的平均加标回收率为83．6％～104％，相对标准偏差为7．2％～13．7％。该方法具有快速简便、设备简单、灵敏度高、准确性好等优点，可很好地应用于酱油中的AFB，的快速检测。

Super pararnagnetic particles with carboxyl group activated by the active ester method, coupled with anti-aflatoxin BI monoclonal antibody purified by the caprylic acid-ammonium sulfate method to form immunomagnetic beads. The enrichment efficiency of immunomagnetic beads of different sizes, and the conditions for antibody coupling and aflatoxin B1 enrichment were studied. A method was established to detect aflatoxin BI in sauce by using an immunomagnetic bead system for aflatoxin BI enrichment and an ELISA method for quantification. The immunomagnetic bead system was compared with the extraction method from the national standard GBfr 5009.22--2003. The ELISA method showed a good linear relationship in the aflatoxin B1 concentration of 0.05 to 0.3 lag/kg （r2 = 0.9842）. Average recovery rates at spiked levels of 1-7 lag/kg were 83.6%--104% with relative standard deviation of 7.2%-13.7%. With the advantages of rapid detection, easy operation, simple instrumental requirements, high sensitivity, and high accuracy, this method can be applied to detect aflatoxin B1 in sauce samples.