title: An HPLC method for the measurement of 5-fluorouracil in human plasma with a low detection limit and a high extraction yield.
abstract: High performance liquid chromatographic (HPLC) techniques for the quantification of 5-fluorouracil (5-FU) in human plasma have been reported in the literature, however, a low limit of detection was generally found to result in a comparatively low extraction yield. We have developed a simple, rapid and sensitive HPLC method for the measurement of 5-FU in plasma which provides both a low limit of quantification and a high extraction yield. This method involves the solid phase extraction of 5-FU from a 500 microl plasma sample. The extract is then injected into an HPLC system equipped with a C18 (mu)Bondapak column, and a UV detector set at 260 nm. Ethyl acetate and potassium dihydrogen phosphate are used for the solid phase extraction and the HPLC mobile phase, respectively. This method provides in a good baseline, a sharp and symmetrical peak for 5-FU, and a high resolution between 5-FU and the internal standard. The retention time of 5-FU using this method is 4.7 min with a limit of detection of 5 ng/ml, and an extraction yield of 96.2+/-0.5% (SE). The next injection is possible in 11 min, and the coefficients of variation are 4.2-8.9% for interday precision, and 5.2-10.6% for day-to-day reproducibility. An HPLC method has been developed that has a low limit of detection and a high extraction yield. This technique was successfully applied in a clinical pharmacokinetic study of 5-FU.
authors.affiliation: Ottawa Regional Cancer Centre, Ottawa, Ontario K1H 1C4, Canada. mnass088@uottawa.ca
authors.forename: Mark Adel
authors.initials: MA
authors.lastname: Nassim
authors.forename: Farshad H
authors.initials: FH
authors.lastname: Shirazi
authors.forename: Christine M
authors.initials: CM
authors.lastname: Cripps
authors.forename: Shereeni
authors.initials: S
authors.lastname: Veerasinghan
authors.forename: Matshela J
authors.initials: MJ
authors.lastname: Molepo
authors.forename: Mihail
authors.initials: M
authors.lastname: Obrocea
authors.forename: Diedre
authors.initials: D
authors.lastname: Redmond
authors.forename: Susan
authors.initials: S
authors.lastname: Bates
authors.forename: Diane
authors.initials: D
authors.lastname: Fry
authors.forename: David J
authors.initials: DJ
authors.lastname: Stewart
authors.forename: Rakesh
authors.initials: R
authors.lastname: Goel
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005472:Fluorouracil / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D006801:Humans
chemical_list: D005472:Fluorouracil