Abstract
Abstract A simple, specific and sensitive high-performance liquid chromatographic (HPLC) method was developed for the determination of l-(5-(3,4-dimetoxyphenyl)-pyrazol-3-yl-oxypropyl)-3-[n-methyl-n-[2-(3,4-dimethoxyphenyl)ethyl]amino] propane hydrochloride (KC11346) and two of its metabolites (KC12795, KC12816) in dog plasma. KC11346, KC12795, KC12816 and KC11294 (internal standard) are extracted from dog plasma by methyl t-butyl ether (MTBE) following an alkalization of plasma. MTBE is removed from the extract with a gentle stream of nitrogen at 40°C and reconstituted in 200 μL of mobile phase. Separation of the reconstituted extract is achieved by HPLC on a Zorbax SB-CN column with a mobile phase composed of (56:44) 50 mM sodium acetate/acetonitrile. The analytes were detected by fluorescence detection with a Corion UG-11 cut off filter and an excitation wavelength of 265 nm. The mean retention times of KC12816, KC12795, KC11346, and the internal standard were 4.8, 8.1, 9.0, and 14.4 minutes, respectiv...
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