Applications of capillary electrophoresis with laser-induced fluorescence for analysis of dGMP-BPDE adduct.
DNA adducts are thought to be crucial to the initiation of mutational and carcinogenic processes. Polycyclic aromatic hydrocarbons (PAHs) have been identified as one major source of carcinogenic risk since they can bind to DNA thus forming an adduct. Quantification of this adduct is important because it may correlate to the risk for cancer development. In this study, the adduct formed between 2'-deoxyguanosine 5'-monophosphate and benzo[ a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE) was analyzed by capillary electrophoresis. Both capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) modes with laser-induced fluorescence detection were used for the separation and analysis of DNA adducts. The exploration of capillary electrophoresis in several modes provided different separation mechanisms in which the stereochemical forms of the adduct could be separated. The best result obtained was using a coated fused-silica capillary in Tris-TAPS buffer, which provided high sensitivity with a detection limit of 2.5x10(-9) mol L(-1). MECC separation of the BPDE adduct, although less sensitive, provided an efficient enantioselective separation option.
Instituto de Química de São Carlos, Universidade de São Paulo, Brazil.
Ana Paula
AP
Formenton-Catai
Emanuel
E
Carrilho
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000032:analysis* / Q000737:chemistry*; D015415:Biomarkers; D018736:DNA Adducts / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry; D003848:Deoxyguanine Nucleotides / Q000032:analysis* / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D007834:Lasers; D015394:Molecular Structure; D013050:Spectrometry, Fluorescence
D015415:Biomarkers; D018736:DNA Adducts; D003848:Deoxyguanine Nucleotides; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; C007257:2'-deoxyguanosine 5'-phosphate