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The in vitro metabolism of 6-chloro-butyl phthalide (CBP) was studied by rat hepatic microsomes. The constitution of the incubation system was optimized and a RP-HPLC-DAD method was developed for the on-line analysis of CBP and its metabolites. The difference of the metabolism profiles of CBP by rat liver microsomes induced by phenobarbital (PB) and 3-methylcholanthrene (3-MC) was also compared. TLC, column chromatography and preparative HPLC were used for the isolation and purification of the major metabolites and they were identified as (R/S) gamma-hydroxy-CBP and (R/S) beta-hydroxy-CBP by UV, NMR and MS.
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