title: Column-switching high-performance liquid chromatographic method for the determination of a thymidylate synthase inhibitor, LY231514, an investigational agent for the treatment of solid tumors, in human plasma.
abstract: A reversed-phase, column-switching high-performance liquid chromatographic (HPLC) method is described for the determination of a new thymidylate synthase inhibitor in human plasma. The compound and an internal standard are extracted from plasma using a Certify II solid-phase cartridge. Extracts are evaporated to dryness and the residue is reconstituted with mobile phase buffer. The analytes are separated from polar interferences and buffer salts originating from the elution step on a 4-mm YMC Basic pre-column. The fraction containing the analytes is further separated on a 25-cm YMC Basic column. The analytes are detected by their absorbance at 250 nm. The limit of quantitation is 10 ng/ml. The method is linear from 10 ng/ml to 80 micrograms/ml using three standard curve ranges. Validation studies for all three ranges show the method to be reproducible. The method has been successfully used to support pharmacokinetic studies.
authors.affiliation: Lilly Laboratory for Clinical Research, Indianapolis, IN 46202.
authors.forename: C L
authors.initials: CL
authors.lastname: Hamilton
authors.forename: J A
authors.initials: JA
authors.lastname: Kirkwood
mesh_terms: D000970:Antineoplastic Agents / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005971:Glutamates / Q000097:blood* / Q000493:pharmacokinetics; D006147:Guanine / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D006801:Humans; D007202:Indicators and Reagents; D000068437:Pemetrexed; D013056:Spectrophotometry, Ultraviolet; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors*
chemical_list: D000970:Antineoplastic Agents; D005971:Glutamates; D007202:Indicators and Reagents; D000068437:Pemetrexed; D006147:Guanine; D013940:Thymidylate Synthase