LC/MS Analysis of Nucleotides on SeQuant® ZIC-cHILIC. Effect of Mobile Phase Buffer Concentration
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 or 100 mM ammonium acetate, pH 5.3 with acetic acid in water
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
1125 psi (77 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
Adjusting buffer concentration and the ionic strength can alter retention and selectivity in HILIC. The buffer salts in HILIC mobile phases are important in reducing electrostatic interactions (attraction and repulsion) between charged analytes and the stationary phase.
法律信息
SeQuant is a registered trademark of Merck KGaA, Darmstadt, Germany