Effect of Buffer on HPLC Separation of Buffers

HEPES, CAPS, MES and MOPS are zwitterionic organic chemical buffering agents. These are very polar compounds which are not retained by traditional reverse phase chromatography. These compounds are zwitterions in nature and can be separated by mixed-mode hydrophilic interaction chromatography on Obelisc N column. Retention is achieved by combination of HILIC and ion-exchange mechanisms. These buffering agents do not have UV active groups, but can be analyzed with ESLD, LC/MS, and CAD detection.

Application Column

Obelisc N

SIELC has developed the Obelisc™ columns, which are mixed-mode and utilize Liquid Separation Cell technology (LiSC™). These cost-effective columns are the first of their kind to be commercially available and can replace multiple HPLC columns, including reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion-exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - buffer concentration, buffer pH, and organic modifier concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.

Select options
Application Analytes:
2-Morpholinoethanesulfonic Acid (MES)
4-Morpholinepropanesulfonic Acid (MOPS)
CAPS (3-(Cyclohexylamino)propanesulfonic acid)
HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic Acid)

Application Detection:
ELSD Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.