Determination of 5-fluorouracil in plasma and whole blood by ion-pair high-performance liquid chromatography.

5-Fluorouracil in blood or plasma was determined by extraction and column liquid chromatography. Acetonitrile was added to blood or plasma and the mixture was stirred and centrifuged. Zinc sulfate addition was followed by stirring and centrifuging. The acetonitrile was salted out with ammonium sulfate, and an aliquot was evaporated with nitrogen. The residue was dissolved in mobile phase and chromatographed. The stationary phase was a styrene-divinylbenzene copolymer, and the mobile phase was 10 mM tetrabutylammonium hydroxide-methanol (74:26). 5-Fluorouracil was detected by UV absorption at 266 nm. Time of the assay was less than 30 min. The detection limit was 10 ng/ml and the relative standard deviation was 4 to 10% depending on the concentration.