Combination of high-performance liquid chromatography and thin-layer chromatography separation of five adducted nucleotides isolated from liver resulting from intraperitoneal administration with 7H-dibenzo[c,g]carbazole to mice.
7H-Dibenzo[c,g]carbazole, DBC, is a potent environmental liver carcinogen. Liver DNA from mice treated with DBC exhibited seven distinct DBC-DNA adducts as detected by 32P-postlabeling using multidimensional TLC. To improve quantitation and chemically characterize the adducts, DNA samples were hydrolyzed, 32P-postlabeled and the adducts were separated from the unadducted normal nucleotides on TLC using a D1 solvent, 0.65 M sodium phosphate (pH 6.8). Adducts were eluted from the TLC plates with 4.0 M pyridinium formate, concentrated, resuspended in 50% aqueous methanol and injected onto the HPLC; five individual adduct peaks were resolved and collected by this method. This approach will prove useful to decrease analysis time and improve chemical characterization of tightly clustered DNA adducts generated in vivo.
O'Connor
P
P
Department of Environmental Health, University of Cincinnati Medical Center, OH 45267-0056, USA.
Fremont
S
S
Schneider
J
J
Dowty
H
H
Jaeger
M
M
Talaska
G
G
Warshawsky
D
D
D000818:Animals; D002227:Carbazoles / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D002274:Carcinogens, Environmental / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D018736:DNA Adducts / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D007274:Injections, Intraperitoneal; D008099:Liver / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008813:Mice, Inbred ICR; D009711:Nucleotides / Q000032:analysis* / Q000737:chemistry; D010761:Phosphorus Radioisotopes
D002227:Carbazoles; D002274:Carcinogens, Environmental; D018736:DNA Adducts; D009711:Nucleotides; D010761:Phosphorus Radioisotopes; C013478:7H-dibenzo(c,g)carbazole