HPLC Determination of HEPES on Newcrom AH Column
https://sielc.com/hplc-determination-of-hepes-on-newcrom-ah-columnChromatogram
Description
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.
Method Parameters
| Mobile Phase | H2O- 100% |
|---|---|
| Buffer | Methanesulfonic acid – 10 mM |
| Flow Rate | 1.0 ml/min |
| Detection | Conductivity |
| Class of Compounds | Hydrophilic |
| Analyzing Compounds | HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic Acid) |
HPLC Column Used
Newcrom AH, 4.6 x 150 mm, 5 µm, 100 A, dual ended