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, 4.6×150 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 Column

Primesep S

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Primesep N

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Obelisc N

SIELC has developed the Obelisc™ columns, which are mixed-mode and utilize Liquid Separation Cell technology (LiSC™). These cost-effective columns are the first of their kind to be commercially available and can replace multiple HPLC columns, including reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion-exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - buffer concentration, buffer pH, and organic modifier concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.

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Application Analytes:
Uracil
Uridine

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.