Determination of MK-507, a novel topically effective carbonic anhydrase inhibitor, and its de-ethylated metabolite in human whole blood, plasma, and urine by high-performance liquid chromatography

Abstract

Sensitive methods for the determination of a novel topically effective carbonic anhydrase inhibitor (CAI) I, MK-507, and its de-ethylated metabolite II, in human whole blood, plasma and urine were developed. These methods were based on liquid—liquid extraction of I and II from biological matrices, back extraction into acid, and analysis by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection (252 nm). The assays were fully validated in the concentration range of 5 to 500 ng/ml, and the limit of quantification (LOQ) for I and II, defined as the lowest concentration on the standard curve for which precision (coefficient of variation, C.V.) is < 10%, was 5 ng/ml in whole blood, plasma, and urine. These methods were applied for the analyses of biological fluid samples from a variety of clinical pharmacokinetic studies. In addition, a method in whole blood based on column-switching HPLC with UV detection and with an LOQ of 50 ng/ml was also developed. The switching valve was used to eliminate interferences from late eluting peaks extracted from whole blood. The details of these assays, together with some representative data from a human study, are presented.