Solid-Core Primesep SB
SIELC Technologies continues to develop mixed-mode Columns with Solid-Core particles with the introduction of Primesep SB mixed-mode HPLC Column. Primesep SB combines efficiency of solid-core technology with unique selectivity of reversed-phase anion-exchange stationary phase. Multiple interactions on a single Column offer 2D type chromatography (Fig. 1) as a tool to separate complex mixtures with a variety of compounds possessing opposing properties: hydrophobic and hydrophilic, neutral and ionic, positively charged and negatively charged.
PrimesepSB Column is designed to retain and separate hydrophobic neutral, hydrophobic basic, hydrophobic acidic, and hydrophilic acidic compounds in one run.
Based on the fact that no two molecules have exactly the same hydrophobic and ionic properties, PrimesepSB explores small differences in such properties for enhanced separation of various compounds
Mixed-mode Columns offer much higher capacity and retention than traditional reversed-phase Columns. Since the surface area of solid-core particles is reduced, mixed-mode phases address potential load-ability issues whilst maintaining the selectivity advantages of multiple interactions. Newly optimized ligand density on Primesep SB Column in combination of carefully selected ionic/hydrophobic ratios further increases the capacity of mixed-mode solid-core Columns.
Primesep SB as well as other solid-core mixed-mode Columns are fully mass chromatography compatible. Mobile phases usually consist of various ratios of acetonitrile and H2O, with buffer or acid. Presence of ions in the mobile phase facilitates an ion-exchange mechanism and elution of ionized compounds.
If a mixture contains a hydrophobic acid it is usually not retained on HILIC Column. Primesep SB Column was used for separation and analysis of benzoic, maleic and fumaric acids. Retention for benzoic acid (pKa 5) comes from RP interaction and stronger acids, like maleic (pKa 1.9 and 6.1) and fumaric (pKa 3.03 and 4.44) are retained mostly by ion-exchange interaction (Fig .7)
Unique chemistry of mixed-mode solid-core, also allows one to retain and separate mono-, di- and tri-phosphates of oligonucleosides. These compounds are highly hydrophilic due to the presence of sugar fragments and presence of phosphates. Mono-, di- and triphosphates can be separated in one run or isolated in groups depending on the number of phosphate residues. (Fig. 8) A much higher concentration of the buffer is required to elute triphosphate from the Column. The method remains LC/MS compatible and prep chromatography compatible.
Solid-Core Primesep SB columns are available in all standard dimensions
Column IDs: 22 mm, 10 mm, 4.6 mm, 3.2 mm, 2.1 mm, 1 mm, 0.5 mm, 0.25 mm
Column Lengths: 250 mm, 150 mm, 100 mm, 50 mm, 25 mm, 10 mm
Particles: 2.7 µm
Pores: 100 A