An HPLC method for the measurement of 5-fluorouracil in human plasma with a low detection limit and a high extraction yield.
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.
Nassim
Mark Adel
MA
Ottawa Regional Cancer Centre, Ottawa, Ontario K1H 1C4, Canada. mnass088@uottawa.ca
Shirazi
Farshad H
FH
Cripps
Christine M
CM
Veerasinghan
Shereeni
S
Molepo
Matshela J
MJ
Obrocea
Mihail
M
Redmond
Diedre
D
Bates
Susan
S
Fry
Diane
D
Stewart
David J
DJ
Goel
Rakesh
R
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005472:Fluorouracil / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D006801:Humans
D005472:Fluorouracil