Determination of a novel angiotensin-AT1 antagonist CR 3210 in biological samples by HPLC.

ABSTRACT

A simple and sensitive method for the determination of a new angiotensin-AT(1) antagonist i.e. CR 3210, 4-[4-[(2-ethyl-5,7-dimethylimidazo[4,5-b]pyridin-3-yl)methyl]phenyl]-3-(2H-tetrazol-5-yl)quinoline, is described. The assay was utilised to describe the pharmacokinetic profile of the title compound after intravenous and intraperitoneal administration to Sprague Dawley rats. CR 3210 and the internal standard CR 1505 (loxiglumide, 4-[(3,4-dichlorobenzoyl)amino-5-[(3-methoxypropyl)pentylamino]-5-oxopentanoic acid) were isolated from rat plasma by solid-phase extraction. The sorbent extraction material along with the pH in the conditioning solution and the washing volume were considered pivotal parameters for the optimisation of the procedure. The separations were performed by reversed-phase high-performance liquid chromatography with ultraviolet detection. The samples were injected onto the analytical column (Tracer Extrasil ODS1) and detected at 238 nm, giving a retention time of 6.19 min for CR 3210 and 4.39 min for the internal standard, respectively. The selectivity of the method showed to be satisfactory. The mean recovery of CR 3210 from spiked rat plasma was 80.3 at 1 microg/ml and 79.9 at 2 microg/ml. The lower limit of detection (LOD) was taken as 0.014 microg/ml in plasma samples. The lower limit of quantification (LOQ) was taken as 0.02 microg/ml, the lowest calibration standard using 500 microg rat plasma. The procedures were validated according to international standards with a good reproducibility and linear response from 0.02 to 2 microg/ml. The sensitivity of the method allowed for its application to pharmacokinetic studies. The maximal concentration was detected 5' after the IV administration, whereas no significant absorption was evident after IP administration of CR 3210 to Sprague-Dawley rats. Our study suggests the absence of extensive bio-transformation of the drug in vivo, supported by the evidence that no metabolites were detected in plasma samples.