Rapid and simultaneous determination of capecitabine and its metabolites in mouse plasma, mouse serum, and in rabbit bile by high-performance liquid chromatography

A rapid high-performance liquid chromatography method has been developed for simultaneous determination of capecitabine and its metabolites: 5'-deoxy-5-fluorocytidine (5'-DFCR), 5'-deoxy-5-fluorouridine (5'-DFUR) and 5-fluorouracil (5-FU). 5'-DFCR was synthesized by hydrolyzing capecitabine using commercially available carboxyl esterase (CES) and characterized by NMR, mass spectrometry and elemental analysis. Base-line separations between capecitabine, 5'-DFCR, 5'-DFUR and 5-FU were found with symmetrical peak shapes on a Discovery RP-amide C16 column using 10 mM ammonium acetate at pH 4.0 and methanol as the mobile phase. The retention times of capecitabine, 5'-DFCR, 5'-DFUR and 5-FU were 8.9, 5.0, 5.3 and 3.0 min, respectively. Linear calibration curves were obtained for each compound across a range from 1 to 500 microg ml(-1). The intra- and inter-day relative standard deviations (%RSD) were <5%. A single-step protein precipitation method was employed for separation of the analytes from bio-matrices. Greater than 85% recoveries were obtained for capecitabine, 5'-DFCR, 5'-DFUR and 5-FU from bio-fluids including mouse plasma, mouse serum and rabbit bile.     

Simultaneous determination of vinclozolin and detection of its degradation products in mouse plasma, serum and urine, and from rabbit bile, by high-performance liquid chromatography

A specific high-performance liquid chromatography method has been developed for simultaneous detection of vinclozolin and its degradation products (M1, M2, and M3). The method has been validated according to ICH guidelines and can be extended to quantitation of vinclozolin. A base-line separation of vinclozolin and its degradation products was found with symmetrical peak shapes on an XTerra MS C18 column using 10 mM ammonium bicarbonate at pH 9.2 and acetonitrile as mobile phase. The retention times of vinclozolin, M1, M2, and M3 were 12.8, 8.1, 11.6, and 11.1 min, respectively. A linear calibration curve was obtained across a range from 5 to 200 microM for vinclozolin. The intra- and inter-day relative standard deviations (%RSD) were <1%. Greater than 90% recoveries of vinclozolin from bio-fluids including mouse plasma, serum and urine, and rabbit bile, were obtained in a single step with a single solvent.