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 liqui d chromatography (HPLC) analysis of adenine nucleosides acid nucleotides, e specially in a biological matrix, is difficult using only ultraviolet detec tion. Derivatization coupled with fluorescence detection has been investiga ted as a means of enhancing sensitivity for the reversed-phase HPLC analysi s of 2 '-beta -fluoro-2 ' ,3 ' -dideuxyadenosine (F-ddA), an experimental, acid-stable, anti-AIDS drug. The reaction of chloroacetaldehyde: with the a denine 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. Th ese derivatives give an analytically useful fluorescence emission at 416 nm after excitation at 230, 265, or 275 nm. Derivatization, fluorescence dete ction and reversed-phase chromatography have been optimized for the analysi s of nanomolar concentrations of F-ddA in human plasma. This method has pot ential for the measurement of F-ddA at low concentration and in limited vol ume samples from in vivo biological studies. Q Published by Elsevier Scienc e B.V.