High-performance liquid chromatographic assay with ultraviolet detection for quantification of dihydrofluorouracil in human lymphocytes: application to measurement of dihydropyrimidine dehydrogenase activity.
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.
Déporte-Féty
R
R
Laboratoire de Pharmacocinétique, Centre Renée Gauducheau, Saint Herblain, France. r-deporte-fety@nantes.fnclcc.fr
Picot
M
M
Amiand
M
M
Moreau
A
A
Campion
L
L
Lanoë
D
D
Renée
N
N
Milano
G
G
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D042943:Dihydrouracil Dehydrogenase (NADP); D005472:Fluorouracil / Q000031:analogs & derivatives / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology / Q000378:metabolism*; D010088:Oxidoreductases / Q000097:blood*; D011874:Radiometry; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
C019389:5-fluorodihydrouracil; D010088:Oxidoreductases; D042943:Dihydrouracil Dehydrogenase (NADP); D005472:Fluorouracil