Hydrophilic interaction chromatography of nucleoside triphosphates with temperature as a separation parameter |
| |
Authors: | Johnsen Elin Wilson Steven Ray Odsbu Ingvild Krapp Andreas Malerod Helle Skarstad Kirsten Lundanes Elsa |
| |
Institution: | Department of Chemistry, University of Oslo, Blindern, N 0315 Oslo, Norway. |
| |
Abstract: | Eight deoxynucleoside triphosphates (dNTPs) and nucleoside triphosphates (NTPs): ATP, CTP, GTP, UTP, dATP, dCTP, dGTP and dTTP, were separated with two 15 cm ZIC-pHILIC columns coupled in series, using LC-UV instrumentation. The polymer-based ZIC-pHILIC column gave significantly better separations and peak shape than a silica-based ZIC-HILIC column. Better separations were obtained with isocratic elution as compared to gradient elution. The temperature markedly affected the selectivity and could be used to fine tune separation. The analysis time was also affected by temperature, as lower temperatures surprisingly reduced the retention of the nucleotides. dNTP/NTP standards could be separated in 35 min with a flow rate of 200 μL/min. In Escherichia coli cell culture samples dNTP/NTPs could be selectively separated in 7 0min using a flow rate of 100 μL/min. |
| |
Keywords: | Nucleotides HILIC Temperature Cell culture van’t Hoff plot |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|