Simultaneous determination of 4-methyl-piperazine-1-carbodithioc acid 3-cyano-3,3-diphenylpropyl ester hydrochloride and its major metabolite in rats by HPLC.

A method for the simultaneous determination of TM208 and its major metabolite (sulfine 4-methyl-piperazine-1-carbodithioc acid 3-cyano-3,3-diphenylpropyl ester, TM208-SO) was developed and validated for the first time. The analytes were extracted from plasma samples by liquid-liquid extraction and analyzed using high-performance liquid chromatography. Flunarizine hydrochloride was used as the internal standard. Chromatographic separations were performed on a Diamonsil C(18) analytical column. The mobile phases consisted of 20 mM ammonium acetate adjusted to pH 4.20 with acetic acid (solvent A) and acetonitrile (solvent B). The analytes were detected at 254 nm after linear gradient elution. The flow rate was 0.8 mL/min. Linearity was obtained over the concentration range of 0.104-5.20 microg/mL for TM208 and 0.145-5.80 microg/mL for TM208-SO in rat plasma. The limit of quantification was 0.104 microg/mL for TM208 and 0.145 microg/mL for TM208-SO, respectively. The inter- and intra-day precision was less than 12.8% for TM208 and 14.1% for TM208-SO. And the accuracy was 96.2-111.1% for TM208 and 95.5-108.6% for TM208-SO. This analytic procedure was applied to a pharmacokinetic study of TM208 and TM208-SO in rats, and the pharmacokinetic parameters were calculated.