Correlation of nucleotide incorporation rate and HPLC retention parameters of substituted nucleosides

A series of 5-alkyl, alkenyl, and alkynyl substituted deoxyuridines and their triphosphate derivatives have been synthetized and studied in DNA polymerase reactions. Initial rate of incorporation of the derivatives catalyzed by Klenow fragment DNA polymerase enzyme (E. coli) was measured. High performance liquid chromatographic (HPLC) retention behaviour of the nucleoside derivatives were investigated on silica and reversed-phase stationary phases using various mixtures of ethylacetate ― methanol and methanol ― water mobile phases, respectively