C18 thin-layer chromatographic enhancement of the 32P-postlabeling assay for aromatic or bulky carcinogen-DNA adducts: evaluation of adduct recoveries in comparison with nuclease P1 and butanol methods.
The suitability of C18 reversed-phase thin-layer chromatography (TLC) for enrichment of adducts in the 32P-postlabeling assay was investigated for structurally diverse classes of DNA adducts derived from benzo[a]pyrene, 2-acetylaminofluorene, benzoquinone, safrole, and mitomycin C. The TLC enrichment involved retention of adducts to the C18 phase followed by elution with organic solvent-water. Adduct patterns obtained by the C18 purification were qualitatively similar to those obtained by the nuclease P1 and butanol procedures, the two commonly used enrichment methods. Adduct recoveries by the C18 method varied for different adducts and were significantly lower than those obtained by the other two techniques.
Reddy
M V
MV
Environmental and Health Sciences Laboratory, Mobil Oil Corporation, Princeton, NJ 08543-1029.
D015073:2-Acetylaminofluorene / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001564:Benzo(a)pyrene / Q000032:analysis / Q000378:metabolism; D016227:Benzoquinones / Q000032:analysis / Q000378:metabolism; D000440:Butanols; D002273:Carcinogens / Q000032:analysis* / Q000378:metabolism; D002855:Chromatography, Thin Layer / Q000379:methods*; D004247:DNA / Q000378:metabolism*; D005260:Female; D007553:Isotope Labeling; D008099:Liver / Q000737:chemistry; D008168:Lung / Q000737:chemistry; D051379:Mice; D016685:Mitomycin / Q000032:analysis / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D051381:Rats; D012451:Safrole / Q000032:analysis / Q000378:metabolism; D015719:Single-Strand Specific DNA and RNA Endonucleases
D016227:Benzoquinones; D000440:Butanols; D002273:Carcinogens; D010761:Phosphorus Radioisotopes; D001564:Benzo(a)pyrene; C004532:quinone; D016685:Mitomycin; D004247:DNA; D015073:2-Acetylaminofluorene; D015719:Single-Strand Specific DNA and RNA Endonucleases; D012451:Safrole