High-performance liquid chromatographic assay with ultraviolet detection for quantification of dihydrofluorouracil in human lymphocytes: application to measurement of dihydropyrimidine dehydrogenase activity
| Autor: | M. Picot, N. Renée, D. Lanoë, Régine Deporte-Fety, Anne Moreau, M. Amiand, Loïc Campion, Gérard Milano |
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| Rok vydání: | 2001 |
| Předmět: |
Lymphocyte
Metabolite High-performance liquid chromatography chemistry.chemical_compound Biotransformation medicine Dihydropyrimidine dehydrogenase Humans Lymphocytes Radiometry Chromatography High Pressure Liquid Dihydrouracil Dehydrogenase (NADP) chemistry.chemical_classification Chromatography Chemistry Reproducibility of Results General Chemistry medicine.anatomical_structure Enzyme Biochemistry Fluorouracil Spectrophotometry Ultraviolet Oxidoreductases Quantitative analysis (chemistry) medicine.drug |
| Zdroj: | Europe PubMed Central |
| ISSN: | 0378-4347 |
| DOI: | 10.1016/s0378-4347(01)00359-0 |
| Popis: | The anticancer drug 5-fluorouracil (5FU) undergoes extensive biotransformation to 5-dihydrofluorouracil (5FUH2) by the enzyme dihydropyrimidine deshydrogenase (DPD). A new HPLC method with direct UV detection for the determination of 5FUH2 in peripheral lymphocytes has been developed to detect DPD deficiency in patients treated with 5FU-based therapy. The method has been shown to be valid over the 5FUH2 concentration range of 1.14-37.88 nmol/ml. Optimal enzymatic conditions for DPD activity measurement were studied: incubation time, protein and 5FU concentrations. The assay was successfully cross-validated with the existing method using HPLC with radiochemical detection. |
| Databáze: | OpenAIRE |
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