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Simultaneous determination of YM‐64227, a phosphodiesterase type 4 inhibitor, and its five metabolites in dog plasma by high‐performance liquid chromatography with fluorescence detection
Author(s) -
Tenmizu Daisuke,
Fukunaga Yasuhisa,
Noguchi Kiyoshi,
Kamimura Hidetaka
Publication year - 2004
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.371
Subject(s) - chemistry , chromatography , analyte , fluorescence , ethyl acetate , metabolite , fluorescence spectroscopy , high performance liquid chromatography , detection limit , quantitative analysis (chemistry) , ether , biochemistry , organic chemistry , physics , quantum mechanics
We developed and validated a reversed‐phase high‐performance liquid chromatographic method with uorescence detection for the simultaneous determination of YM‐64227 [4‐cyclohexyl‐1‐ethyl‐7‐methylpyrido(2,3‐d)pyrimidin‐2‐(1 H )‐one], a novel and selective phosphodiesterase type 4 inhibitor, and its ve hydroxylated metabolites in dog plasma. The plasma samples were extracted with tert‐butyl methyl ether under alkali conditions. The analytes were well separated on a phenyl ethyl column (5 µm, 250 × 4.6 mm i.d.), opreating at 40°C and using an acetonitrile–acetic acid gradient at a ow rate of 1.0 mL/min. The uorescence signal was monitored at an excitation and emission wavelength of 330 and 400 nm, respectively. No interfering peak was observed at the retention time of YM‐64227, its metabolites or the internal standard. The validated quantitation range of the method was 0.4–200 ng/mL for all analytes using 0.5 mL of the plasma sample. The recovery of analytes in the extraction process was more than 65.5%. The intra‐ and inter‐assay precision was less than 5.1 and 12.6%, respectively, and the intra‐ and inter‐assay accuracy ranged from ‐8.1 to 11.8% and ‐8.0 to 9.9%, respectively. Using this assay, the plasma concentration of YM‐64227 and metabolites can be determined after the oral administration of YM‐64227 to beagle dogs. Copyright © 2004 John Wiley & Sons, Ltd.