COMBINATION OF HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AND THIN-LAYER CHROMATOGRAPHY SEPARATION OF 5 ADDUCTED NUCLEOTIDES ISOLATED FROM LIVER RESULTING FROM INTRAPERITONEAL ADMINISTRATION WITH 7H-DIBENZO[C,G]CARBAZOLE TO MICE

7H-Dibenzo[c,g]carbazole, DEC, is a potent environmental liver carcino gen. Liver DNA from mice treated with DEC exhibited seven distinct DBC -DNA adducts as detected by P-32-postlabeling using multidimensional T LC. To improve quantitation and chemically characterize the adducts, D NA samples were hydrolyzed, P-32-postlabeled and the adducts were sepa rated 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 p lates with 4.0 M pyridinium formate, concentrated, resuspended in 50% aqueous methanol and injected onto the HPLC; five individual adduct pe aks were resolved and collected by this method. This approach will pro ve useful to decrease analysis time and improve chemical characterizat ion of tightly clustered DNA adducts generated in vivo. (C) 1997 Elsev ier Science B.V.