Improved reversed-phase high-performance liquid chromatographic separation of 32P-labelled nucleoside 3',5'-bisphosphate adducts of polycyclic aromatic hydrocarbons.
32P-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 32P-labelled 3',5'-bisphosphates of nucleosides modified by reactive derivatives of carcinogenic polycyclic aromatic hydrocarbons (PAH). Optimal resolution of the major 32P-postlabelled DNA adducts formed by the anti-diol-epoxides of ten PAH was achieved using a phenyl-modified 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.
Pfau
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Haddow Laboratories, Institute of Cancer Research, Sutton, Surrey, UK.
Phillips
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DH
D002273:Carcinogens / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004247:DNA / Q000737:chemistry / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds / Q000737:chemistry*; D011874:Radiometry
D002273:Carcinogens; D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds; D004247:DNA