E. Gamelin et al., A SIMPLE CHROMATOGRAPHIC METHOD FOR THE ANALYSIS OF PYRIMIDINES AND THEIR DIHYDROGENATED METABOLITES, Journal of liquid chromatography & related technologies, 20(19), 1997, pp. 3155-3172
Citations number
21
Categorie Soggetti
Chemistry Analytical","Biochemical Research Methods
Fluorouracil (5-FU) is a pyrimidine analog widely used in the treatmen
t of numerous malignancies. One major metabolic pathway is the reducti
on of its double bond by dihydropyrimidine dehydrogenase (DYPD), whose
activity is a strong determinant of 5-FU plasma levels. Recent findin
gs show that deficiencies in DYPD are less rare than generally assumed
and play a major role in 5-FU toxic effects. We developped a simple,
sensitive and accurate liquid chromatographic method that allows the s
imultaneous determination of uracil, 5-fluorouracil and their dihydrog
enated metabolites in plasma. This method offers a useful tool for the
detection of defects in pyrimidine degradation. HPLC was carried out
by using Spherisorb ODS1 (10 cm) and ODS2 (25 cm) columns serially mou
nted, and 10 mM phosphate buffer, pH 3.0, as the mobile phase with UV
detection at 205 nm. Many parameters, such as mobile phase pH, ionic s
trength, column temperature, were found to have a marked influence on
the results. We calculated the ratio dihydrouracil/uracil and could de
scribe a Gaussian distribution of this ratio in a population of 78 hea
lthy volunteers.