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Simultaneous determination of flucytosine and fluorouracil in human plasma by high‐performance liquid chromatography
Author(s) -
Sastre Toraño J.,
Vermes A.,
Guchelaar H.J.
Publication year - 2001
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.38
Subject(s) - chemistry , chromatography , repeatability , calibration curve , trichloroacetic acid , reproducibility , high performance liquid chromatography , detection limit , analytical chemistry (journal)
A validated, sensitive and precise reversed‐phase high‐performance liquid chromatographic method for the simultaneous determination of 5‐flucytosine (5‐FC) and 5‐fluorouracil (5‐FU) in human plasma is described. Two compounds, 5‐methylcytosine (5‐MC) and 5‐chlorouracil (5‐CU), were used as internal standards for the determination of 5‐FC and 5‐FU, respectively. Plasma samples were deproteinized with trichloroacetic acid and chromatographed on an octylsilica column, maintained at 30°C during elution, using a 0.04 M phosphate buffer, pH 7.0, as eleunt. Spectrophotometric diode array detection was used at 266 nm. 5‐FC, 5‐FU, 5‐MC and 5‐CU were found to have retention times of 4.8, 5.8, 7.7 and 11.0 min respectively. Recoveries of 91–120% with reproducibility and repeatability coefficients of variation of 0.8–6% were obtained. Mean correlation coefficients of 0.99989 and 0.9995 were found for the linear calibration curves ( n = 2) of 5‐FC (4.816–192.6 mg/l) and 5‐FU (0.05368–5.368 mg/l), respectively. The limits of quantitation were 0.3 mg/l for 5‐FC and 0.05 mg/l for 5‐FU. Copyright © 2001 John Wiley & Sons, Ltd. Abbreviations used: 5‐CU 5‐chlorouracil5‐FC 5‐flucytosineFdUMP 5‐fluorodeoxyuridine monophosphate5‐FU 5‐fluorouracilFUTP 5‐fluorouridine triphosphateHLQ higher limit of quantitationLLQ lower limit of quantitation5‐MC 5‐methylcytosineTDM therapeutic drug monitoring.