Evaluation of a fluorogenic derivatization method for the reversed-phase HPLC analysis of 2'-beta-fluoro-2',3'-dideoxyadenosine, a new anti-AIDS drug.
High sensitivity (10(-7) to 10(-9) M) reversed-phase high-performance liquid chromatography (HPLC) analysis of adenine nucleosides and nucleotides, especially in a biological matrix, is difficult using only ultraviolet detection. Derivatization coupled with fluorescence detection has been investigated as a means of enhancing sensitivity for the reversed-phase HPLC analysis of 2'-beta-fluoro-2',3'-dideoxyadenosine (F-ddA), an experimental, acid-stable, anti-AIDS drug. The reaction of chloroacetaldehyde with the adenine base has been employed to form fluorescent 1,N(6)-etheno derivatives of F-ddA and 5'-deoxyadenosine, which is used as an internal standard. These derivatives give an analytically useful fluorescence emission at 416 nm after excitation at 230, 265, or 275 nm. Derivatization, fluorescence detection and reversed-phase chromatography have been optimized for the analysis of nanomolar concentrations of F-ddA in human plasma. This method has potential for the measurement of F-ddA at low concentration and in limited volume samples from in vivo biological studies.
Zhang
H
H
Laboratory of Medicinal Chemistry, Division of Basic Sciences, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, MD 21702-1201, USA.
Ford
H
H
Roth
J S
JS
Kelley
J A
JA
D019380:Anti-HIV Agents / Q000032:analysis* / Q000097:blood; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D016048:Dideoxyadenosine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D012015:Reference Standards; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D019380:Anti-HIV Agents; D007202:Indicators and Reagents; C073072:lodenosine; D016048:Dideoxyadenosine