LC/MS Analysis of Nucleotides on SeQuant® ZIC-HILIC. Effect of pH

LC/MS Analysis of Nucleotides on SeQuant(R) ZIC-HILIC. Effect of pH


instrument Agilent 6530 Q-TOF LC/MS
column SeQuant ZIC-HILIC, 10 cm x 2.1 mm, 3.5 µm
mobile phase   [A] acetonitrile; [B] 50 mM ammonium acetate, pH 4.5, 5.0, 5.3, or 5.5 with acetic acid in water, pH 6.9 not adjusted
gradient 26 to 27% B in 10 min; to 27 to 35% B in 10 min; to 26% B in 0.1 min; hold at 26% B for 4.9 min
flow rate 0.3 mL/min
pressure 1088 psi
column temp. 40 °C
detector Q-TOF (ESI-negative), XIC of the following m/z values: AMP-346, ADP-426, ATP-506, CMP-322, CDP-402, CTP-482, GMP-362,  GDP-442,  GTP-522, UMP-323, UDP-403, UTP-483
injection 10 µL


Featured Industry Life Science and Biopharma
Legal Information SeQuant is a registered trademark of Merck KGaA, Darmstadt, Germany
Analysis Note ZIC-HILIC is a densely bonded zwitterionic stationary phase with sulfobetaine functional groups graft-polymerized to silica. The stationary phase has an overall 1:1 charge balance between the permanent positive quaternary ammonium groups close to the silica support and the distal negatively charged sulfonate groups. It has though a slight negative surface charge because of the spatial arrangement. A covalently bonded zwitterionic stationary phase, with permanent strong acidic and basic functionalities, provide hydrophilic partitioning combined with weak ionic interactions and makes the phase virtually pH independent as shown in this analysis.
Application No. G1007625


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