Determination of 5-Fluorouracil and Its Prodrug Tegafur in Plasma and Tissue by High-Performance Liquid Chromatography in a Single Injection: Validation for Application in Clinical Pharmacokinetic Studies

Abstract

Tegafur, a prodrug of 5-fluorouracil (5-FU), is an oral fluorouracil antitumor drug used for the management of adenocarcinomas. It has an efficacy similar to that of intravenous 5-FU, with potential advantages in terms of convenience and quality of life for the patient and cost-effectiveness as compared with intravenous chemotherapy. The authors developed a high-performance liquid chromatography (HPLC) assay for the determination of tissue or plasma tegafur and 5-FU concentration in a single step extraction and a single HPLC injection. The retention times of 5-FU and tegafur were 5 and 16.5 minutes, respectively, and the internal standard retention times were 11.5 and 17.5 minutes for 5-bromouracil (5-BU) and beta-hydroxyethyltheophylline, respectively. The limit of quantification was 0.0125 microg/mL for 5-FU and 0.05 microg/mL for tegafur. The assay had good recovery (96.5% +/- 9.45% and 97.5% +/- 7.89% for 5-FU in plasma and tissue, respectively, and 88.5% +/- 12.17% and 104.9% +/- 8.77% for tegafur in plasma and tissue, respectively). Precision was good: the within-day and between-day standard deviation of the mean (RSD) for 5-FU (0.0125-5 microg/mL) and tegafur (0.5-150 microg/mL) was always <8%. The authors conclude that the method described here is ideally suited for the therapeutic monitoring of 5-FU and tegafur.

Determination of 5-fluorouracil in human plasma by high-performance liquid chromatography (HPLC)

“…Furthermore, the method required only 0.5 mL of plasma, with the LLOQ of 5 ng/mL. Compared with recent reports, the sensitivity obtained from our method was similar to those of Matsushima et al [16] , Chu et al [17] and Remaud et al [23] and better than those of Zufia et al [14] , Maring et al [15] and Bocci et al [19] . The method can be applied to pharmacokinetic studies of 5-FU in humans and improve response/toxicity balance by therapeutic drug monitoring.…”

“…Various methods for the determination of 5-FU in biological matrix have been reported. These methods employ gas chromatography, tachophoresis or highperformance liquid chromatography (HPLC) as separating technologies, with radioactivity, nitrogen-phosphorus flame-ionization, ultraviolet, fluorescence or mass spectrometry as detection techniques [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] . Unfortunately, most of these methods required laborious sample preparation involving derivatization step before analysis, long runtime and complex apparatus.…”

Extraction sur phase solide et analyses par HPLC du 5-fluoro-uracile plasmatique

“…Les chromatogrammes obtenus après extraction du plasma sur phase solide montrent que la nouvelle technique d'extraction proposée dans ce travail est beaucoup plus sensible, dans la mesure où les limites de détection et de quantification du 5-FU sont très abaissées par rapport aux résultats d'autres chercheurs (la limite de quantification du 5-FU est de 12,5 ng ml -1 pour Zufia et al [13] et de 40 ng ml -1 pour Maring et al) [14]. Même avec 2,5 ng, on observe nettement le pic du 5-FU (Fig.…”

Simple determination of capecitabine and its metabolites by liquid chromatography with ultraviolet detection in a single injection

“…The published HPLC methods for 5-FU determination are relatively sensitive but also time-consuming and sometimes require long and involved extraction procedures [20][21][22][23][24][25][26][27][28][29]. A simple and sensitive HPLC method for determination of 5-FU in plasma was developed in our laboratory [30] and another simple and rapid HPLC method (also optimized and validated in our laboratory) for simultaneous quantification of both 5-FU and its fluoropyrimidine prodrug Tegafur in human plasma and tissue has recently been published [31].…”