NMR-STUDY OF WHOLE RAT BILE - THE BILIARY-EXCRETION OF 4-CYANO-N,N-DIMETHYL ANILINE BY AN ISOLATED-PERFUSED RAT-LIVER AND A LIVER IN-SITU

The structure of two biliary metabolites of 4-cyano-N,N,-dimethyl aniline (CDA) contained in whole rat bile have been studied in detail by NMR at 400 MHz. A 4-cyano-N-methyl glutathione-N-aniline conjugate was identified as a biliary metabolite of CDA using relatively simple IH NMR techniques. Isotopically C-13 labelled CDA was used to generate C-13 labelled xenobiotic conjugates. Our use of H-1/C-13 heteronuclear NMR techniques, in particular a C-13-selective HMQC-TOCSY experiment, allowed a N-beta-glucuronide conjugate, a previously unknown biliary metabolite of CDA, to be identified. Bile samples obtained from both the isolated perfused rat liver and the rat liver in situ were analysed.