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An Optimized Ion-Pair HPLC Method for Simultaneous Analysis of Nucleoside Triphosphate Levels in Hepatoma Cell Line
Authors:Jing Jia  Hai Zhang  Liang Zhao  Zhen-yu Zhu  Guo-qing Zhang  Yi-feng Chai
Institution:1. Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai, 200433, People??s Republic of China
3. Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, 200438, People??s Republic of China
2. Shanghai Key Laboratory for Pharmaceutical Metabolite Research, No. 325 Guohe Road, Shanghai, 200433, People??s Republic of China
Abstract: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 ± 3.49% to 103.1 ± 1.75%, respectively. Finally, the method was successfully applied to the analysis of eight compounds present in HepG2 cell extracts.
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