HEPES is a zwitterionic organic chemical buffering agent. It is a very polar compound which is not retained by traditional reverse phase chromatography. HEPES can be separated by mixed-mode hydrophilic interaction chromatography on the Newcrom AH mixed-mode column, which has both hydrophobic and cationic exchange properties. The mobile phase consists of H2O and Methanesulfonic acid. It can be detected using a conductivity detector.
Column | Newcrom AH, 4.6×150 mm, 5 µm, 100A |
Mobile Phase | H2O- 100% |
Buffer | Methanesulfonic acid – 10 mM |
Flow Rate | 1.0 ml/min |
Detection | Conductivity |
Class of Compounds |
Hydrophilic |
Analyzing Compounds | HEPES |
Application Column
Newcrom AH
The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.
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