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Journal of Analytical Methods in Chemistry
Volume 2013, Article ID 584964, 8 pages
Research Article

Fluorimetric Immunoassay for Multianalysis of Aflatoxins

Biosensor Lab., Department of Chemistry, BITS, Pilani-K. K. Birla Goa Campus, Goa 403726, India

Received 16 April 2013; Revised 27 June 2013; Accepted 28 June 2013

Academic Editor: Lingzhi Liu

Copyright © 2013 Lizy Kanungo and Sunil Bhand. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


A sensitive fluorimetric ELISA was developed for the analysis of aflatoxins. The assay was performed in a 384 microwell plate, wherein high specificity monoclonal antibody against AFM1 (mAb-AFM1) was used as capture antibody and FITC conjugated secondary antibody was used for detection and quantification of the analyte. The linear range of the immunoassay was found to be 6.25–50 pg/mL. AFM1 as low as 1 pg/mL was detected by this method with assay volume 40 μL. The multi-analysis of different aflatoxins was also investigated in the microwell plate, based on the cross-reactivity (CR) approach. Real milk samples were tested along with certified reference material by standard addition method and recovery analysis was done. The mAb-AFM1 showed 23.2% CR with AFB1, 50% CR with respect to AFM2, and least CR towards AFG1 (<1%). Furthermore, mixture analysis of AFM2 and AFB1 was carried out at specific concentrations of AFM1. The advantages of this developed immunoassay are high sensitivity, high throughput, multianalyte detection, versatility, and ease of handling.