Methods for the Detection and Characterization of Metabolites of the Potential Cytostatic Drug Oracine

Barbora Szotáková (szotako@faf.cuni.cz), Vladimír Wsól, Eva Kvasničková

Department of Biochemical Sciences, Faculty of Pharmacy, Charles University, 500 05 Hradec Králové, Czech Republic

Summary

In order to study the metabolism of oracine, a potential cytostatic drug, in vitro methods for preparing oracine metabolites (substrate concentration, incubation period, selection of coenzymes, aerobic and anaerobic incubation), conditions for extracting non-metabolised substrate and of its metabolites from various biological samples (enzyme incubates, urine and faeces samples from experimental animals administered with oracine) as well as methods for their chromatographic separation and detection (TLC and HPLC) have been developed and optimised. Three oxidised metabolites and a product of oracine reduction have been detected in vitro and compared with the in vivo metabolites.