Hydrophilic Interaction Chromatography
Hydrophilic Interaction Chromatography - Related Videos
03:07
Hydrophilic Interaction Liquid Chromatography: A Technique to Separate Hydrophilic Polar Analytes Using Hydrophilic Beads
0 Views •
2025
This video demonstrates the principle of separation of hydrophilic polar compounds using hydrophilic interaction liquid chromatography. This technique helps in the purification of various polar analytes, including N-glycans.
03:22
Calcium-Dependent Hydrophobic Interaction Chromatography: A Technique to Purify Calcium-Binding Proteins Based on Hydrophobic Interactions
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2025
In this video, we demonstrate the purification of calcium-binding protein from a dialyzed cell lysate through calcium-dependent hydrophobic interaction chromatography. The calcium-binding proteins expose a hydrophobic region upon binding with calcium, facilitating interaction with a hydrophobic group on resin. Later these proteins are eluted using calcium chelator EDTA that reverses the interaction.
10:21
Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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Cited by 17 •
2011
An automated method for identifying suitable hydrophobic interaction chromatography (HIC) media to be used in the process of protein purification is presented. The method utilizes a medium-pressure liquid chromatography system including automated buffer blending, dynamic sample loop injection, sequential column selection, multi-wavelength analysis, and split fraction eluate collection.
03:46
Butyl-Dependant Hydrophobic Interaction Liquid Chromatography: A Technique to Characterize Antibody-Drug Conjugates Based on Hydrophobic Interactions
0 Views •
2025
This video demonstrates the hydrophobic interaction liquid chromatography-based characterization of antibody-drug conjugates. Using a butyl ligand-based chromatography column, the antibody-drug conjugates are characterized via hydrophobic interactions with the ligand.
09:01
A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating
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2016
This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic. The produced nanoparticle is an assembly of gliadin-cyanoacrylate diblock copolymers. Spray coating with the produced nanoparticle changes the surface of target material to a hydrophilic surface.
FAQs
The polar stationary phase supports a water-enriched layer at its surface even though the mobile phase is rich in organic solvent. Polar analytes interact with this interfacial layer and can also participate in hydrogen bonding and electrostatic interactions. Because these mechanisms contribute alongside partitioning, compounds with different polar characteristics can show distinct retention and separation behavior.
An organic-rich mobile phase, typically containing acetonitrile, creates the conditions needed for the surface water layer and provides selectivity for highly polar compounds. This complements reversed-phase chromatography, where such compounds may be poorly retained. The contrast allows investigators to choose HILIC when polar biomolecules require a different separation environment from that provided by traditional reversed-phase methods.
Retention reflects several interactions rather than a single property. Partitioning between the mobile phase and the water-enriched surface layer contributes to separation, while hydrogen bonding and electrostatic interactions further distinguish analytes. Their combined effects help resolve chemically diverse polar compounds, including metabolites, carbohydrates, glycopeptides, and nucleotides, in complex biological samples.
A HILIC analysis requires a polar stationary phase and an organic-rich mobile phase, typically based on acetonitrile, so the interfacial water-enriched layer can form. The sample is then assessed according to how its polar constituents interact with that environment. This combination provides the separation conditions needed to examine multiple classes of polar biomolecules within complex biological material.
It is particularly useful when a study targets highly polar compounds that are poorly retained by traditional reversed-phase chromatography. Biological applications include metabolomics, glycomics, pharmaceutical research, and biomarker studies. The method can address analytes such as metabolites, carbohydrates, glycopeptides, and nucleotides, making it relevant when biological interpretation depends on measuring diverse polar molecular features.
Its compatibility with mass spectrometry links chromatographic separation with sensitive molecular analysis of polar compounds. By separating constituents before detection, HILIC can provide complementary selectivity for complex biological samples and broaden the range of features examined. This combination is valuable in metabolomics, glycomics, pharmaceutical research, and biomarker studies where polar biomolecules are central to the investigation.