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Updated: Oct 26, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Simultaneous determination of 5-fluorouracil and uracil by high-performance liquid chromatography using four serial
L K House1, J Ramírez, M J Ratain
1Department of Medicine, The University of Chicago, IL 60637, USA.
Abstract:
A sensitive assay was developed for the quantitation of 5-fluorouracil (5-FU) and uracil using liquid-liquid extraction (LLE) and HPLC with UV detection. Analyses were performed with four microBondapak C18 columns connected in series using 20 mM acetic acid with 1% ACN as mobile phase. The calibration curves were linear across the range of 26-1000 ng ml(-1) (0.21-7.8 microM) for 5-FU and 1.0-14.0 microg ml(-1) (0.01-110 microM) for uracil. This assay has been implemented to determine the plasma concentrations for pharmacokinetic studies for 5-FU and uracil in conjunction with clinical trials.
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