An Optimized Ion-Pair HPLC Method for Simultaneous Analysis of Nucleoside Triphosphate Levels in Hepatoma Cell Line

An improved ion-pair HPLC method was developed for the simultaneous determination of ribonucleoside triphosphates and their corresponding deoxyribonucleoside triphosphates in HepG2 cell extracts. HPLC conditions, flow rate and column temperature were optimized and good linearity (r (2) > 0.9993) was obtained over the investigated concentration ranges. Reproducibility was evaluated by intra- and inter-day assays and RSD values were below 5.39%. Recoveries ranged from 98.2 +/- A 3.49% to 103.1 +/- A 1.75%, respectively. Finally, the method was successfully applied to the analysis of eight compounds present in HepG2 cell extracts.