title: High-performance liquid chromatography of 5-fluorouracil after derivatization with 4-bromomethyl-7-methoxycoumarin. Characterization of the derivative and the use of column switching for the improvement of resolution and the enhancement of sensitivity.
abstract: The derivatization of 5-fluorouracil with 4-bromomethyl-7-methoxycoumarin has been reported previously; however, the structure of the derivative was not confirmed. The synthesis and purification of the 5-FU derivative is described along with the spectroscopic (MS and NMR) determination that it is labelled at both heterocyclic nitrogens as expected. A column switching HPLC system is also presented which consists of primary separation on a cyanopropyl column followed by a final separation on an ODS column with fluorescence detection. This system removes all interferences from the derivatization system and has a limit of detection for the pure derivative of less than 50 fmol (injection volume = 100 microliters).
authors.affiliation: Center for Bioanalytical Research, University of Kansas, Lawrence 66045.
authors.forename: C G
authors.initials: CG
authors.lastname: Kindberg
authors.forename: M
authors.initials: M
authors.lastname: Slavik
authors.forename: C M
authors.initials: CM
authors.lastname: Riley
authors.forename: J F
authors.initials: JF
authors.lastname: Stobaugh
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation; D002853:Chromatography, Liquid; D005472:Fluorouracil / Q000032:analysis* / Q000737:chemistry; D015394:Molecular Structure; D013053:Spectrophotometry; D014468:Umbelliferones / Q000737:chemistry*
chemical_list: D014468:Umbelliferones; C000317:4-bromomethyl-7-methoxycoumarin; D005472:Fluorouracil