Immunoaffinity Columns

IACs are prepared by adsorbing immunospecific antibodies onto a gel support contained in a plastic column or cartridge (131). The sample extract is loaded onto the column, and as the extract is forced through, the analyte is captured by the antibodies. Impurities not bound to the antibodies are washed from the cartridge. Mycotoxins are then desorbed with methanol and quantitated by HPLC or by fluorometry with derivitization, if needed. Advantages of IAC over ELISA methods include selectivity, the ability to trap toxins from large volumes of sample extract, and the ability to combine with different analytical techniques. The cost of columns can be high (>$10.00 each), which can make analysis of many samples expensive (129). However, several reports have indicated that IAC could be reused, in some cases more than 100 times (58-60,129). Several investigators used IAC for analyzing several toxins simultaneously. Maragos et al. (62) developed a single IAC that simultaneously isolated fumonisin Bj and B2 and hydro-lyzed forms of these toxins. In contrast, Scudamore et al. (61) linked two IACs in series to purify aflatoxins and ochratoxin A.

IACs are available from commercial sources for cleanup of aflatoxins B1( B2, G^ G2 and M1( deoxynivalenol, fumon-isins Bx and B2, ochratoxin A, and zearalenone. Antibodies against other mycotoxins (citrinin, patulin, cyclopiazonic acid, kojic acid, sterigmatocystin, and several others) have been produced. However, no reports have indicated the use of IAC for purification of these mycotoxins (129). IAC purification and SPE gave generally similar results for aflatoxins (154) in milk, corn, and sorghum, but higher recoveries were obtained when IACs were used to isolate fumonisin Bx from canned and frozen corn (155).

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