Amide Hydrophilic Chromatography
Amide Hydrophilic Chromatography - Related Videos
03:07
Hydrophilic Interaction Liquid Chromatography: A Technique to Separate Hydrophilic Polar Analytes Using Hydrophilic Beads
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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.
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A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating
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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.
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Preparation of Amides
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2023
Amides are synthesized by treating carboxylic acids with amines in the presence of dehydrating agents like dicyclohexylcarbodiimide (DCC). The DCC-promoted synthesis of amides begins with the protonation of DCC by carboxylic acid. The protonation makes it a better acceptor. Next, the addition of carboxylate to the protonated carbodiimide gives a reactive acylating agent. Subsequently, the amine acts as a nucleophile that attacks the acylating agent to form a tetrahedral intermediate. In the...
01:19
Amines to Amides: Acylation of Amines
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2025
Various carboxylic acid derivatives (such as acid chlorides, esters, and anhydrides) can be used for the acylation of amines to yield amides. The reaction requires two equivalents of amines. The first amine molecule functions as a nucleophile and attacks the carbonyl carbon to produce a tetrahedral intermediate. This is followed by the loss of the leaving group and restoration of the C=O bond. Next, the second equivalent of amine serves as a Brønsted base and deprotonates the quaternary amide...
01:28
Amides to Carboxylic Acids: Hydrolysis
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2023
Amides can undergo either acid-catalyzed hydrolysis or base-promoted hydrolysis through a typical nucleophilic acyl substitution. Each hydrolysis requires severe conditions. Acid-catalyzed hydrolysis: Hydrolysis of amides under acidic conditions yields carboxylic acids. Since the reaction occurs slowly, hydrolysis requires the conditions of heat. The mechanism begins with the protonation of the carbonyl oxygen by the acid catalyst. The protonation makes the amide carbonyl carbon more...
FAQs
Retention reflects more than simple polarity. A water-enriched layer forms at the polar amide stationary-phase surface, and analytes partition between this layer and the organic-rich mobile phase. Hydrogen bonding and other hydrophilic interactions further influence how strongly each compound is retained. Differences in these interactions allow highly polar biomolecules to elute at distinct times.
The organic-rich mobile phase creates a contrast with the water-enriched environment associated with the stationary-phase surface. Polar analytes therefore distribute between two chemically different environments rather than remaining uniformly in the mobile phase. Their differing affinity for the surface-associated water layer and mobile phase produces retention differences that enable chromatographic resolution.
The principal distinction is the type of compounds each approach can resolve effectively. Highly polar metabolites, carbohydrates, and related biomolecules may perform poorly in conventional reversed-phase chromatography, whereas the amide hydrophilic format is designed to retain and separate them through surface-associated water, hydrogen bonding, and other hydrophilic interactions. This expands coverage of biological molecules.
Analyte polarity is a central factor, but retention also depends on the compound's ability to participate in hydrogen bonding and other hydrophilic interactions. Its partitioning between the organic mobile phase and water-enriched stationary-phase layer contributes as well. Consequently, structurally different biomolecules with similar overall polarity can still show distinct retention and separation behavior.
The technique is applied to biological samples to separate polar constituents before molecular measurement. Its resolving power helps distinguish metabolites, carbohydrates, and other biomolecules within complex extracts. In many workflows, the chromatographic separation is combined with mass spectrometry, so separated compounds can support molecular identification and quantitative analysis rather than being assessed only as unresolved mixtures.
It is particularly useful when a study targets highly polar compounds that are inadequately resolved by conventional reversed-phase methods. In metabolomics, it can improve separation of metabolite constituents; in glycomics, it supports analysis of carbohydrates and related biomolecules. The same capabilities also assist targeted biomarker measurements in complex biological extracts.