HPLC Analysis of Nucleosides on Ascentis® Express OH5 (HILIC Mode)

HPLC Analysis of Nucleosides on Ascentis® Express OH5 (HILIC Mode)

Conditions

column Ascentis Express OH5, 10 cm x 2.1 mm I.D., 2.7 µm particles (53757-U)
column temp. 25 C
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
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)
injection 2 µl
detector UV, 250 nm

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.
Categories Analytical Chromatography, Nucleic Acids
Featured Industry Life Science and Biopharma
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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