LC/MS Analysis of Nucleotides SeQuant® ZIC-cHILIC. Effect of pH.
CONDITIONS
column
SeQuant ZIC-cHILIC, 10 cm x 2.1 mm, 3.0 μm particles (< href="/ProductLookup.html?ProdNo=150657&Brand=MM">1.50657.0001)
mobile phase
[A] acetonitrile; [B] 50 mM ammonium acetate, pH as indicated with acetic acid in water (50 mM ammonium acetate, pH 6.93 in water 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 9.9 min
flow rate
0.3 mL/min
pressure
986 psi (68 bar)
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
sample
10-30 μg/mL of nucleotides (AMP, ADP, ATP, CMP, CDP, CTP, GMP, GDP, GTP, UMP, UDP, UTP) (80/20 acetonitrile/50 mM ammonium acetate in water pH 5.3 with acetic acid)
说明
分析说明
ZIC-cHILIC is a densely bonded zwitterionic stationary phase with phosphorylcholine functional groups covalently attached to silica. The stationary phase has an overall 1:1 charge balance between the permanent negative phosphate groups close to the silica support and the distal positively quaternary ammonium groups. It has though a slight positive surface charge because of the spatial arrangement. A true 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.
法律信息
SeQuant is a registered trademark of Merck KGaA, Darmstadt, Germany