HILIC Retention of Uridine and Uracil on Sielc’s HILIC Columns

Uridine and uracil are separated on three HILIC columns from SIELC. Primesep N, Primesep S, and Obelisc N columns all have different polarity and different embedded acidic groups on the surface of silica gel: Primesep N is a normal-phase HPLC column with embedded acidic groups with a pKa of about 5. Primesep S is a normal-phase HPLC column with embedded acidic groups with a pKa of about 3. The Primesep S stationary phase retains basic compounds by cation-exchange at pH > 3. In traditional HILIC mode, a charged or neutral polar analyte interacts with a water layer on the polar stationary phase surface. On Obelisc N the charges are greatly separated and independently accessible, resulting in different selectivity compared to traditional HILIC and silica columns. Mobile phase composition changes the conformation of the long hydrophilic chain. All three columns can be used to analyze uridine and uracil by HPLC with UV, LC/MS and ELSD detection.

Condition

Column Primesep S, Primesep N, Obelisc N, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O - 95/5%
Buffer AmAc pH 4.0 - 5mM
Flow Rate 1.0 ml/min
Detection UV 270 nm
 

Description

Class of Compounds Nucleosides,  Hydrophilic, Ionizable
Analyzing Compounds Uracil, Uridine
 

Application Analytes:

Uracil
Uridine