Abstract
The adenine base of 2′,3′-dideoxyadenosine-5′-triphosphate (ddATP) was chosen as the target for precapillary derivatization for its analysis by CE with fluorescence detection. Several fluorophores, such as chloroacetaldehyde, 9-fluorenylmethyloxycarbonylchloride (FMOC), fluorescamine (Fluram), o-phthaldialdehyde (OPA) and complexation with terbium (Tb), gadolinium (Gd), and phenanthroline (phen) were tried. Precapillary derivatization of ddATP with chloroacetaldehyde showed poor peak shape for the ε-ddATP adduct and degradation peaks of ε-adenine, possibly due to the low sample pH of 4.0. Precapillary derivatization of ddATP with OPA/β-mercaptoethanol, Fluram, and FMOC showed no fluorescent derivatization products. Complexation of ddATP with Tb-Gd-phen produced a sharp fluorescent peak. The sensitivity of the Tb-Gd-ddATP-phen complex was found to be 20 times higher than UV detection of ddATP.
ACKNOWLEDGMENTS
Funding of this work by the Flemish Fund for Scientific Research (Research Project G.0355.06) is gratefully acknowledged. S.M thanks the onderzoeksfonds/K. U. Leuven for financial support.