[Determination of new H2-receptor antagonist Z-300 in plasma by high-performance liquid chromatography with fluorescence detection].

Abstract

A highly specific, sensitive and reproducible high-performance liquid chromatographic method was developed for the determination of a new H2-receptor antagonist, N-[3-[3-(piperidinomethyl)phenoxy]propyl]-2-(2- hydroxyethylthio)acetamide.2-(4-hydroxybenzoyl)benzoate(Z-300), in rat, dog and human plasma, Z-300 base was isolated from the plasma using a Extrelut-1 solid-phase extraction cartridge. The Z-300 base was converted to a fluorescent derivative by the use of 1-anthroyl cyanide in the presence of quinuclidine as catalyst. The derivatized mixture was treated with Sep-pak CN Vac cartridge to remove interferences from endogenous substances. The derivative obtained was separated on a Inertsil ODS-2 column using a acetonitrile-0.2% ammonium acetate-methanol (7:6:6) containing 0.5 mM dodecyltrimethylammonium bromide (ion-pair reagent) as a mobile phase with a fluorescence detector. The detection limit of Z-300 base in the plasma was 0.5 ng/ml. The calibration curves were linear over the range of 2-200 ng/ml. The method appears to be readily applicable to the study of Z-300 pharmacokinetics in animals and humans.