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: M Qian, P J O'Dwyer, J M Gallo
• Journal: Journal of chromatography. B, Biomedical applications (J Chromatogr B Biomed Appl) Vol. 666 Issue 2 Pg. 307-14 (Apr 21 1995) ISSN: 1572-6495 [Print] Netherlands
• PMID: 7633607 (Publication Type: Clinical Trial, Clinical Trial, Phase I, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antimetabolites, Antineoplastic
• Indoles
• AG 331
• Thymidylate Synthase

Topics

• Antimetabolites, Antineoplastic (blood, pharmacokinetics, urine)
• Chromatography, High Pressure Liquid (methods)
• Humans
• Indoles (blood, pharmacokinetics, urine)
• Reproducibility of Results
• Spectrum Analysis
• Thymidylate Synthase (antagonists & inhibitors)