title: Hydrophilic interaction chromatography of nucleoside triphosphates with temperature as a separation parameter.
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.
authors.affiliation: Department of Chemistry, University of Oslo, Blindern, N 0315 Oslo, Norway.
authors.forename: Elin
authors.initials: E
authors.lastname: Johnsen
authors.forename: Steven Ray
authors.initials: SR
authors.lastname: Wilson
authors.forename: Ingvild
authors.initials: I
authors.lastname: Odsbu
authors.forename: Andreas
authors.initials: A
authors.lastname: Krapp
authors.forename: Helle
authors.initials: H
authors.lastname: Malerod
authors.forename: Kirsten
authors.initials: K
authors.lastname: Skarstad
authors.forename: Elsa
authors.initials: E
authors.lastname: Lundanes
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D012265:Ribonucleotides / Q000737:chemistry / Q000302:isolation & purification*; D013696:Temperature
chemical_list: D012265:Ribonucleotides