A cost effective preparative thin layer chromatography cleanup method for high performance liquid chromatography analysis of aflatoxins B1, B2 and G2

Naveed Ahmed, Sohail Hassan Khan, Muhammad Ashraf Anjum, Abdul Rehman


Aflatoxins are the by-products of fungal metabolism and common contaminants in feed. To keep their level below permissible limits, various assays have been developed. Currently, High Performance Liquid Chromatography (HPLC) with fluorescence detection of toxins is most widely used method but the cleanup methods requiring specific gadgets have increased the cost of the assay. Thin Layer Chromatography (TLC) is effective and economical but it only gives semi quantitative determination for aflatoxin.

This study explores the preparative potential of TLC as cleanup method for HPLC analysis of toxin. Standard aflatoxins solutions containing 10 ng of B1, 2.5 ng of B2 and 2.5 ng of G2 were spotted on silica plates and then extracted using chloroform and acetone (4:1). The extracted toxins were resolved and quantified on HPLC using fluorescent detection. The results showed 81, 79 and 60% mean recovery of aflatoxin B1, B2 and G2, respectively. This method was proved equivocally comparable to other methods of aflatoxins cleanup and thus can be used as an alternative cost effective cleanup method.

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