IMPROVED REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC SEPARATION OF P-32 LABELED NUCLEOSIDE 3',5'-BISPHOSPHATE ADDUCTS OF POLYCYCLIC AROMATIC-HYDROCARBONS

P-32-Postlabelling is a sensitive technique for the detection and analysis of carcinogen-DNA adducts. In this paper we describe the development of an improved high-performance liquid chromatography (HPLC) method for the separation of P-32-labelled 3',5'-bisphosphates of nucleosides modified by reactive derivatives of carcinogenic polycyclic aromatic hydrocarbons (PAH). Optimal resolution of the major P-32-postlabelled DNA adducts formed by the anti-diol-epoxides of ten PAH was achieved using a phenylmodified silica gel column with a gradient of methanol in phosphate buffer at low pH and high ionic strength. Use of a radioactivity flow detector coupled to the HPLC apparatus allowed detection of subfemtomole quantities of labelled adducts.