HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF CYTOSINE-CONTAINING COMPOUNDS BY PRECOLUMN FLUORESCENCE DERIVATIZATION WITH PHENACYL BROMIDE - APPLICATION TO ANTIVIRAL NUCLEOSIDES AND NUCLEOTIDES
Ej. Eisenberg et Kc. Cundy, HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF CYTOSINE-CONTAINING COMPOUNDS BY PRECOLUMN FLUORESCENCE DERIVATIZATION WITH PHENACYL BROMIDE - APPLICATION TO ANTIVIRAL NUCLEOSIDES AND NUCLEOTIDES, Journal of chromatography B. Biomedical applications, 679(1-2), 1996, pp. 119-127
Citations number
13
Categorie Soggetti
Chemistry Analytical","Biochemical Research Methods
Journal title
Journal of chromatography B. Biomedical applications
A novel precolumn derivatization method for the HPLC determination of
cytosine-containing compounds by HPLC is described. Highly fluorescent
2-phenyl-3,N-4-ethenocytosine derivatives are produced by a reaction
of non-fluorescent cytosine-containing compounds with phenacyl bromide
in weakly acidic acetonitrile solution at elevated temperature. The a
pplicability of the method to various biogenic and antiviral compounds
is demonstrated. Quantitative determination of cidofovir, a potent an
tiviral drug currently undergoing evaluation in the clinic for treatme
nt of cytomegalovirus retinitis is also reported. The limit of detecti
on for cidofovir in cynomolgus monkey plasma was 5 ng/ml (ca. 100 fmol
on column) with the between-day precision of 16.6, 6.4 and 2.4% for f
ive replicate samples at 20, 160 and 320 ng/ml, respectively. The with
in-day precision was 15.9, 5.9 and 2.1%, respectively. The method desc
ribed has broad applicability and may offer significant advantages ove
r existing HPLC methods in antiviral drug development as well as in nu
cleic acid research.