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Immunomagnetic Bead Enrichment and ELISA for Detection of Aflatoxin B1 in Sauce

XIE Fang,LAI Wei-hua,SHI Ai-wu,DENG Sheng-liang,XIONG Yong-hua,YU Yang-fan   

  1. 1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;
    2. Wuxi Zodolabs Biological Technology Co. Ltd., Wuxi 214000, China;
    3. Institute of Microbiology, Jiangxi Academy of Sciences, Nanchang 330029, China
  • Online:2013-09-25 Published:2013-09-27

Abstract:

Super paramagnetic particles with carboxyl group activated by the active ester method, coupled with anti-aflatoxin B1
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 B1 in sauce by using an immunomagnetic bead system for aflatoxin B1 enrichment
and an ELISA method for quantification. The immunomagnetic bead system was compared with the extraction method from the
national standard GB/T 5009.22—2003. The ELISA method showed a good linear relationship in the aflatoxin B1 concentration
of 0.05 to 0.3 μg/kg (r2 = 0.9842). Average recovery rates at spiked levels of 1–7 μg/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.

Key words: immunomagnetic beads, sauce, aflatoxin B1 (AFB1), ELISA method