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What is HILIC?
? Hydrophilic Interaction Liquid Chromatography (HILIC) was first described by Alpert*.
? HILIC is ideal for the separation and retention of polar species including polar neutral and polar ionised analytes.
? HILIC separations typically include a polar stationary phase with high organic solvent containing mobile phases.
? Mechanistically HILIC is complex (Fig 1) and provides multiple modes of interaction between the analyte,stationary phase, eluent and water enriched layer at the stationary phase

particle-eluent interface**.
* A. J. Alpert, J. Chromatogr., 499 (1990) 177.
** See the FREE ACE guide to reproducible HILIC method development for more information – order your copy now.

When Should You Consider HILIC?
? HILIC provides the retention and separation of hydrophilic or polar to very polar analytes not well retained in RPLC.
? Hydrophilic or polar to very polar analytes have log P values (measure of lipophilicity) of around zero or less (Fig 2A).
? Generally, polar analytes are suitable for HILIC if they elute before caffeine in gradient RPLC (Fig 2B).

HILIC Provides Alternative Selectivity and Retention to RPLC

? HILIC provides improved retention and alternative selectivity to RPLC for polar and very polar analytes.
? Analysis of a six component polar analyte mixture by RPLC and HILIC shows inverted elution profiles (see below).

Conclusions
ACE HILIC columns provide alternative selectivity to ACE RPLC columns with excellent retention of polar analytes.

ACE HILIC Columns - 3 Options
Three ACE HILIC stationary phases have been developed to provide the solution to your HILIC separation needs.

To extend column lifetime under HPLC conditions (up to 5000 psi / 350 bar), ACE guard cartridges (5/pk) or ACE pre-column filters (5/pk) are recommended. For the
guard system a guard cartridge holder (H0001) and coupler (C0001) are also required. To extend column lifetime under UHPLC conditions (up to 15000 psi / 1000 bar) an ACE UHPLC pre-column filter is recommended.

ACE HILIC-A
? An acidic character phase with an ionisable negative surface charge depending upon mobile phase pH.

ACE HILIC-B
? A basic character phase with an ionisable positive surface charge depending upon mobile phase pH.

ACE HILIC-N
? A polar neutral character phase with the capability of H-bonding amongst other mechanisms of interaction.

ACE HILIC Method Development
? ACE HILIC columns provide alternative selectivity to each other.
? The power of systematic screening of different phase chemistries for HILIC method development is seen below.
? Maximise your HILIC method development success by following the ACE HILIC method development protocol using three optimised ACE HILIC column chemistries – protocol available in a FREE HILIC guide.

ACE HILIC UHPLC / HPLC and Microbore Columns Part Numbers
Matched guard cartridges (for HPLC columns) or prefilters (for UHPLC columns) are available – please enquire.