HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ASSAY FOR 3'-AMINO-3'-DEOXYTHYMIDINE IN PLASMA, WITH 9-FLUORENYL METHYL CHLOROFORMATE AS THE DERIVATIZATION AGENT
H. Nattouf et al., HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ASSAY FOR 3'-AMINO-3'-DEOXYTHYMIDINE IN PLASMA, WITH 9-FLUORENYL METHYL CHLOROFORMATE AS THE DERIVATIZATION AGENT, Journal of chromatography B. Biomedical applications, 664(2), 1995, pp. 365-371
Citations number
8
Categorie Soggetti
Chemistry Analytical
Journal title
Journal of chromatography B. Biomedical applications
We have developed a sensitive high-performance liquid chromatographic
assay for the determination of the zidovudine metabolite 3'-amino-3'-d
eoxythimidine (AMT) using fluorescence detection and sensitivity in th
e picomolar range. Plasma was diluted with 0.05 M sodium phosphate buf
fer pH 7.2 and subsequently prepared for analysis using solid-phase ex
traction. AMT was derivatized with 9-fluorenyl methylchloroformate and
chromatographed using a reversed-phase system. The mobile phase consi
sted of acetonitrile-0.01 M potassium phosphate buffer (pH 7) (32:68,
v/v). The fluorescence of the column effluent was monitored at 262 nm
(excitation) and 306 nm (emission). Good resolution of AMT from endoge
nous plasma components was obtained. Within- and between-day variabili
ty was less than 10%. The limit of quantitation was 0.9 mu g/l. The as
say was successfully applied to the determination of AMT in human plas
ma and in plasma of mice treated with zidovudine.