HPLC Analysis of Nucleosides on Ascentis® Express OH5 (HILIC Mode)
Materials
analytical column
standard
CONDITIONS
column
Ascentis Express OH5, 10 cm x 2.1 mm I.D., 2.7 μm particles (53757-U)
mobile phase
[A] 5 mM ammonium acetate, pH 5.0 with acetic acid in 95:5, acetonitrile:water; [B] 5 mM ammonium acetate, pH 5.0 with acetic acid in 80:20, acetonitrile:water
gradient
0% B held for 1 min; to 100% B in 10 min; held at 100% B for 1 min
flow rate
0.3 mL/min
column temp.
25 °C
detector
UV, 250 nm
injection
2 μL
sample
10 - 100 μg/mL in 95:5, acetonitrile:water (pH of buffer stock (in water) was adjusted before further dilution with water and/or acentonitrile)
Description
Analysis Note
Nucleosides are involved in many aspects of metabolism. Analogs of nucleosides have medical application in antivirals or anticancer treatments. This application demonstrates the utility of hydrophilic interaction chromatography (HILIC) for the retention and resolution of nucleosides and analogs thereof. Elution is not solely explained by logD but by other ionic and solvophobic effects.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany