EFFICIENT CONJUGATION OF AFLATOXIN B1 WITH BOVINE SERUM ALBUMIN APPLYING FOR DEVELOPMENT OF AFLATOXIN B1 QUICK TEST
Keywords:Aflatoxin B1, Aflatoxin B1- BSA conjugate, quick test
AbstractAflatoxins are secondary metabolites mostly produced by Aspergillus flavus and Aspergillus parasiticus and found in agricultural foodstuff such as maize grains, peanuts, animal feeds,... These are toxic and cancerous agents for humans and animals. Among them, aflatoxin B1 is the most consideration due to its highest toxicity and presence in samples. Several methods were developed to detect aflatoxin in food, feed and other foodstuffs such as chromatographic methods (TLC, HPTLC, HPLC), LC-MS/MS, FTIR, RIA, ELISA, SPR, electrochemical, and immunodipstick. Among them only immunodipstick method is compatible for field usage. Therefore, aim of the present study is to determine optimal condition for conjugation of aflatoxin B1 (AFB1) with bovine serum albumin (BSA) to develop a lateral flow immunoassay test strip for detection of aflatoxin B1. Optimal conditions for generating mediate compound of aflatoxin B1 – CMO was investigated: AFB1:CMO ratio of 1:2, reflux temperature of 100oC for 2 hours. Concentration of aflatoxin B1 in reaction was 0.27 µg/µl. Conjugation of AFB1 with CMO was confirmed by thin layer chromatography, HPLC and FT-IR methods. Furthermore, optimal conditions for conjugation of mediate AFB1-CMO derivative with BSA were also investigated. Ratio of AFB1-CMO with BSA was determined as 40:1 and equal to predicted theoretical ratio. Efficient conjugation conditions were 25oC for 2 hours in bicarbonate buffer pH 9.5. Generated conjugate of AFB1-BSA was successfully applied to construct a lateral flow immunoassay test strip for detection of aflatoxin B1
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