Abstract
Administration of 5-fluorouracil (5-FU) may be associated with severe toxicities in patients who are deficient of dihydropyrimidine dehydrogenase (DPD) activity. For this reason, a sensitive HPLC method for the analysis of 5-FU and 5-fluoro-5, 6-dihydrouracil (5-FDHU) was developed in the present study for the determination of DPD activity in nucleated cells of peripheral blood and pharmacokinetic analysis of 5-FU and 5-FDHU in humans. 5-FU and 5-FDHU were extracted from biologic matrices by adding sodium acetate, sodium sulfate, and diethyl ether/propanol. Dried samples were reconstituted in a mobile phase (KH2PO4 35 mmol/L, pH 4.0), isocratically eluted with a Hypersil C18 stationary phase (25 cm × 4.6 mm, 10 μm), and detected by a diode array detector (measurement and reference wavelengths, 215 and 360 nm, respectively). 5-Fluorocytosine (internal standard), 5-FDHU, and 5-FU were eluted within 13 minutes of the injection without interferences. Recoveries ranged between 81% to 85% for all compounds, and the method proved to be linear, with a coefficient of linearity of 0.999. The limits of detection and quantification were 3.2 and 16 ng/mL, respectively, and the within-day and between-day CV were less than 10% for both 5-FU and 5-FDHU. The present assay proved to be sufficiently sensitive and specific to evaluate cellular DPD activity and measure 5-FU and 5-FDHU plasma concentrations in cancer patients, thus allowing therapeutic 5-FU monitoring in patients and identification of DPD-deficient subjects at major risk of severe toxicities.
Original language | English |
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Pages (from-to) | 362-368 |
Number of pages | 7 |
Journal | Therapeutic Drug Monitoring |
Volume | 27 |
Issue number | 3 |
DOIs | |
Publication status | Published - Jun 2005 |
Keywords
- 5-fluorouracil
- Dihydropyrimidine dehydrogenase
- Diode array detector
- Pharmacokinetics
- Therapeutic drug monitoring
- Toxicity
ASJC Scopus subject areas
- Toxicology
- Health, Toxicology and Mutagenesis
- Pharmacology
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Pharmacology (medical)
- Public Health, Environmental and Occupational Health