Hydrophilic interaction chromatography of nucleoside triphosphates with temperature as a separation parameter
Hydrophilic interaction chromatography of nucleoside triphosphates with temperature as a separation parameter
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DOI：
10.1016/j.chroma.2011.01.066
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发表时间：
2011-09-02
影响因子：
4.1
通讯作者：
Lundanes, Elsa
中科院分区：
文献类型：
--
作者：
Johnsen, Elin;Wilson, Steven Ray;Lundanes, Elsa
用两个串联的15 cm ZIC-pHILIC色谱柱，使用LC-UV仪器分离八种脱氧核苷三磷酸（dNTP）和核苷三磷酸（NTP）：ATP、CM、GTP、UTP、dATP、dCTP、dGTP和dTTP。基于聚合物的ZIC-pHILIC柱比基于二氧化硅的ZIC-HILIC柱提供了显著更好的分离和峰形。与梯度洗脱相比，等度洗脱获得更好的分离。温度显著影响选择性，可用于微调分离。分析时间也受温度影响，因为较低的温度令人惊讶地降低了核苷酸的保留。dNTP/NTP标准品可在35分钟内以200 μ L/min的流速分离。在大肠杆菌细胞培养样品中，dNTP/NTP可在70分钟内以100 μ L/min的流速选择性分离。（C）2011 Elsevier B. V.保留所有权利。
Eight deoxynucleoside triphosphates (dNTPs) and nucleoside triphosphates (NTPs): ATP, CM, 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 mu L/min. In Escherichia coli cell culture samples dNTP/NTPs could be selectively separated in 70 min using a flow rate of 100 mu L/min. (C) 2011 Elsevier B.V. All rights reserved.