DETECTION OF HUMAN EXPOSURE TO CARCINOGENS BY MEASUREMENT OF ALKYL-DNA ADDUCTS USING IMMUNOAFFINITY CLEANUP IN COMBINATION WITH GAS-CHROMATOGRAPHY MASS-SPECTROMETRY AND OTHER METHODS OF QUANTITATION

A brief overview is given of recent developments from our laboratory i n the use of immunoaffinity clean-up in the determination of alkyl-DNA adducts. Compound- and group-specific antibodies have been prepared a gainst 7-alkylguanines and 3-alkyladenines. The antibodies were attach ed to solid supports to make immunoaffinity columns which could then b e used to selectively purify either single adducts or groups of adduct s prior to quantitation by various methods. In the case of methyl addu cts quantitation was achieved by ELISA (3-methyladenine, using a monoc lonal antibody) and HPLC-electrochemical detection (7-methylguanine). For groups of adducts, quantitation of the individual compounds was ef fected by gas chromatography-mass spectrometry (3-alkyladenines, using deuterated analogues of each adduct as an internal standard) and HPLC -fluorescence detection (7-alkylguanines). In all of these cases effic ient purification of adducts from urine or DNA hydrolysates could be e asily carried out. Using these techniques human exposure to alkylating agents in tobacco smoke and from cancer chemotherapy has been studied .