HILIC SEP R TIONS WITH H LO®

Hydrophilic interaction liquid chromatography (HILIC)
is a useful UHPLC and HPLC mode for the following
situations:
• Polar analytes that are poorly or not retained in
RPLC
• Basic analytes that have poor peak shape
(overloading) and/or poor retention at low pH in
RPLC
• Analytes that have log P values near or less than
zero
HALO ® HILIC is a Fused-Core silica phase that can be
used either in HILIC mode or in normal-phase mode
with water-immiscible solvents (NPLC).
HALO ® Penta-HILIC is a bonded silica phase, which
has a highly polar ligand with 5 hydroxyl groups
tethered via novel proprietary linkage chemistry to
Fused-Core silica particles.
Some Typical Analytes for HILIC Separations
• Basic pharmaceuticals

• When conditions orthogonal to RPLC mode are • Peptides

needed (elution order change) • Polar organic acids
• Catecholamines and other neurotransmitters
HALO® columns are currently available in two different
phases for HILIC separations: • Nucleosides and nucleobases
• HALO® HILIC • Drug glycoside and glycuronide metabolites
• HALO® Penta-HILIC • Mono-, di-, tri- and other oligosaccharides
• Opiates
• Glycosylceramides
• Polar triazines and pyrimidines
• nalytes from metabolomic pro ling

For more information on HILIC separations, please see

references 7-10 on page 31.

RETENTION ORDER REVERSAL AND IMPROVED RETENTION WITH HILIC

adenosine +
cytosine
acenaphthene

Figure R.
You can o ten obtain TEST CONDITIONS:
a complete reversal Column: HALO® 90 Å C18, 2.7 µm, 4.6 x 50 mm
Absorbance mAU

Part Number: 76444-964

in elution order and Mobile Phase A: 90/10 ACN/0.1 M Ammonium Formate
di erent selectivity cytosine Flow Rate: 1.8 mL/min.
adenosine pH: 3.0
using HILIC mode
acenaphthene
compared to TEST CONDITIONS:
Column: HALO® 90 Å HILIC, 2.7 µm, 4.6 x 50 mm
reversed phase Part Number: 76444-962
mode under the Mobile Phase A: 90/10 ACN/0.1 M Ammonium Formate
Flow Rate: 1.8 mL/min
same or appropriate pH: 3.0
conditions.

0 0.5 1 1.5

Time min.

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