| CAS Number | 7365-45-9 |
|---|---|
| Molecular Formula | C8H18N2O4S |
| Molecular Weight | 238.301 |
| InChI Key | JKMHFZQWWAIEOD-UHFFFAOYSA-N |
| LogP | -2.26 |
| Synonyms |
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Applications:
Uv-Vis Spectrum of HEPES
January 30, 2026
If you are looking for optimized HPLC method to analyze HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic Acid) check our HPLC Applications library
For optimal results in HPLC analysis, it is recommended to measure absorbance at a wavelength that matches the absorption maximum of the compound(s) being analyzed. The UV spectrum shown can assist in selecting an appropriate wavelength for your analysis. Please note that certain mobile phases and buffers may block wavelengths below 230 nm, rendering absorbance measurement at these wavelengths ineffective. If detection below 230 nm is required, it is recommended to use acetonitrile and water as low UV-transparent mobile phases, with phosphoric acid and its salts, sulfuric acid, and TFA as buffers.
For some compounds, the UV-Vis Spectrum is affected by the pH of the mobile phase. The spectra presented here are measured with an acidic mobile phase that has a pH of 3 or lower.
HPLC Determination of HEPES on Newcrom AH Column
October 20, 2020
HPLC Method for HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic Acid) on Newcrom AH by SIELC Technologies
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 x 150 mm, 5 µm, 100 A, dual ended |
| 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) |
Application Column
Newcrom AH
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Column options: dual ended
Effect of Buffer on HPLC Separation of Buffers
August 22, 2008
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 options4-Morpholinepropanesulfonic Acid (MOPS)
CAPS (3-(Cyclohexylamino)propanesulfonic acid)
HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic Acid)
Zwitterion
