06/17/2009 • Analytics

NUCLEODUR® HILIC - A useful complement to conventional HPLC phases

Separation science is always looking for new and effective strategies to accomplish the tasks of modern analytics. Especially for polar compounds reversed phase HPLC – the most common analytical method – is often limited. Here, hydrophilic stationary phases provide an additional tool for the separation of polar analytes in HPLC.

The expression HILIC (Hydrophilic Interaction LIquid Chromatography) was firstly published by Andrew Alpert in 1990 – since then it took quite some efforts to develop robust and reproducible hydrophilic HPLC phases for HILIC chromatography [A. Alpert, J. Chromatography 499 (1990), 177–196].
HILIC combines the characteristics of the 3 major methods in liquid chromatography – reversed phase (RPC), normal phase (NPC) and ion chromatography (IC):

”HILIC is NP chromatography of polar and ionic compounds under RP conditions.”

Surface modification
NUCLEODUR® HILIC is a special zwitterionic modified stationary phase based on ultra spherical NUCLEODUR® particles. The betaine character of the ammonium-sulfonic acid ligands results in total charge equalisation and in an overall neutrally charged but highly polar surface.

Retention charcteristic
Commonly HILIC is described as partition chromatography or liquid/liquid extraction system between the mobile and stationary phase. Versus a water-poor mobile phase a water-rich layer on the surface of the polar stationary phase is formed. Thus, a distribution of the analytes between these two layers will occur.
Furthermore HILIC includes weak electrostatic mechanisms as well as hydrogen donor interactions between neutral polar molecules under high organic elution conditions.

Key features of NUCLEODUR® HILIC

  • ideal for reproducible and stable chromatography of highly polar analytes
  • suitable for analytical and preparative applications as well as LC-MS
  • very short column conditioning and equilibration period

Recommended application
hydrophilic compounds such as organic polar acids and bases, polar natural compounds, nucleosides, oligonucleotides, amino acids, peptides, water soluble vitamins

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