HPLC Analysis of Polar Compounds on Ascentis® Express OH5 (HILIC Mode)
CONDITIONS
column
Ascentis Express OH5, 10 cm x 3 mm I.D., 2.7 μm particles (53769-U)
mobile phase
5 mM ammonium formate (95:5 acetonitrile:water) pH 6.8
flow rate
0.6 mL/min
pressure
965 psi (66.5 bar)
column temp.
30 °C
detector
UV, 254 nm
injection
1 μL
sample
100 μg/mL in 25:75, water:methanol
Description
Analysis Note
The impact of partitioning and ion exchange in HILIC separations is demonstrated using a mixture of acidic, neutral, and basic polar compounds in this Figure. All three analytes have similar log D values, both acid and base are ionized at mobile phase pH.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany