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Qian MX , Odwyer PJ , Gallo JM
High-Performance Liquid-Chromatographic Method for the Determination of Ag-331, a Novel Anticancer Agent, in Human Serum and Urine Using Solid-Phase Extraction and Photodiode- Array Detection
Journal of Chromatography B-Biomedical Applications. 1995 Apr 21;666(2) :307-314
PMID: ISI:A1995QW51200013   
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A reversed-phase isocratic high-performance liquid chromatographic method has been developed for the determination of AG-331, a novel thymidylate synthase inhibitor, in human serum and urine. The method involves a solid-phase extraction from C-18 cartridges without addition of an internal standard. The methanol eluent is evaporated under nitrogen at 40 degrees C, and reconstituted in mobile phase, acetonitrile-water (35:65, v/v) containing 25 mM ammonium phosphate. Separation of AG-331 was obtained on a C-18 column at a flow-rate of 1 ml/min. Chromatographic signals were monitored by a photodiode- array detector at a primary wavelength of 457 nm with a bandwidth of 4.8 nm. Standard curves are linear in the range of 22-2175 ng/ml in plasma and 44-2175 ng/ml in urine, respectively. The extraction recovery ranged from 92.9-102.4%. Intra-day coefficient of variation was less than 9.5%, and inter-day coefficient of variation was less than 14.3% for an AG-331 concentration of 44 ng/ml. This method has been used to characterize the pharmacokinetics of AG-331 in cancer patients as part of ongoing Phase I trials.
Times Cited: 1 Article QW512 J CHROMATOGR B-BIO MED APPL