A sensitive fluorescence assay for quantitation of fludarabine and metabolites in biological fluids.
Rapid and quantitative dephosphorylation of the new anticancer nucleotide analogue fludarabine phosphate to its nucleoside 9-beta-D-arabinofuranosyl-2-fluoroadenine (F-ara-A) renders this metabolite the target for pharmacologic investigations. At clinically effective doses of fludarabine phosphate (18-30 mg/m2 per day) comprehensive pharmacokinetic analysis of F-ara-A has been limited by the sensitivity of UV based HPLC assays. To address this problem we developed a sensitive test based on the condensation of F-ara-A with chloroacetaldehyde to form the fluorescent derivative, arabinosyl-1,N6-etheno-isoguanine. Combined with a solid-phase extraction step prior to derivatization and separation of the reaction products by reverse-phase HPLC, this assay had a quantitation limit of 2 pmol F-ara-A per ml plasma. Slightly modified, the system was also applicable to urine specimens, with a quantitation limit of 1 nmol F-ara-A per ml urine.
Kemena
A
A
University of Texas M.D. Anderson Cancer Center, Houston 77030.
Fernandez
M
M
Bauman
J
J
Keating
M
M
Plunkett
W
W
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004358:Drug Therapy; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008658:Inactivation, Metabolic; D007262:Infusions, Intravenous; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy*; D012680:Sensitivity and Specificity; D001084:Vidarabine Phosphate / Q000031:analogs & derivatives* / Q000494:pharmacology
D000964:Antimetabolites, Antineoplastic; D005456:Fluorescent Dyes; D001084:Vidarabine Phosphate; C042382:fludarabine phosphate