title: High-performance liquid chromatographic method for the determination of AG-331, a novel anti-cancer agent, in human serum and urine using solid-phase extraction and photodiode-array detection.
abstract: 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 C18 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 C18 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.
authors.affiliation: Department of Medical Oncology, Fox Chase Cancer Center, Philadelphia, PA 19111, USA.
authors.forename: M
authors.initials: M
authors.lastname: Qian
authors.forename: P J
authors.initials: PJ
authors.lastname: O'Dwyer
authors.forename: J M
authors.initials: JM
authors.lastname: Gallo
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007211:Indoles / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D015203:Reproducibility of Results; D013057:Spectrum Analysis; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors
chemical_list: D000964:Antimetabolites, Antineoplastic; D007211:Indoles; C087748:AG 331; D013940:Thymidylate Synthase