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ZORBAX HILIC Plus
InfinityLab Poroshell 120
HILIC
Most common HILIC applications are for the analysis of
polar compounds that are unretained or poorly retained
on reversed-phase columns.
Best to equilibrate the column with 30-40% water in
acetonitrile before use.
Equilibration may require 20-50 column volumes.
A typical mobile phase is acetonitrile:water with an
acetate or formate buffer at 5-10 mM. Increasing buffer
concentration can improve peak shape and retention.
Decrease the aqueous/buffer component to increase
retention.
HILIC Method Development Notes
Operating Guidelines: HILIC
• Agilent HILIC columns are shipped containing acetonitrile:water or acetonitrile.
• Agilent HILIC columns are compatible with water and all common organic
solvents.
• All silica has some solubility in pH >6 aqueous mobile phases. Solubility of the
silica is also increased at elevated temperatures. Using the columns above pH 6
and 40 °C will reduce the column lifetime.
Column Name pH Range Maximum
Temperature
Suggested
Temperature
ZORBAX HILIC Plus
InfinityLab Poroshell 120
HILIC
pH 1.0 to 8.0 40 °C 30 °C
ZORBAX Carbohydrate pH 2.0 to 8.0 40 °C 30 °C
HILIC Column Operating Parameters
Operating at extreme ends of pH and temperature ranges will have a significant impact
on column lifetime.
As acetonitrile content of the mobile phase increases, the resolution of the sugars
may improve, with an accompanying increase in analysis time. The typical range
for the mobile phase is 60% to 75% acetonitrile. Resolution may be improved when
column temperature is above room temperature. Operation at 30 °C or 35 °C may
give better resolution of a carbohydrate mixture.
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