Analysis of Cooked-Food Mutagens by HPLC/Immunoassay.

摘要

Recognizing that an immunoassay for these aminoimidazoazaarenes (AIAs) could be an analytical method with higher sample throughput and would be less costly, we developed a set of monoclonal antibodies that selectively bind to each of the AIAs. We selected PhIP (6-phenyl-2-amino-1-methylimidazo(4,5-b)pyridine) for the initial assay development because it is the AIA that is most abundant by mass in cooked meat and it is the most genotoxic AIA in mammalian-cell short-term bioassays. It is, however, the least active AIA in the Ames Salmonella mutagenesis assay. We have recently developed a set of four monoclonals that bind to the PhIP. These antibodies were produced by standard methods and were derived from the immunogen described previously. The binding specificity of each of these antibodies has been well characterized. The most specific antibody, called PhIP-1, will bind PhIP with a 50% inhibition point (I sub 50 ) of 30 ng, and it will not bind to any of the other AIA mutagens, nor to any of a number of synthetically produced derivatives of PhIP, such as Iso-PhIP (6-phenyl-2-amino-3-methylimidazo(4,5-b)pyridine). PhIP-1 does bind with 2-deamino-PhIP and 2-deamino-2-nitro-PhIP with an I sub 50 of 13 and 16 ng, respectively. 9 refs., 3 figs. (ERA citation 14:003588)