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Related Concept Videos

Mass Spectrometry of Amines01:15

Mass Spectrometry of Amines

In mass spectroscopy, amines undergo fragmentation to give parent ions with odd molecule weights. This observed mass spectrum follows the nitrogen rule; a molecule with an odd number of nitrogen atoms produces a molecular ion with an odd molecular weight. Amines undergo fragmentation through α cleavage, producing nitrogen-containing cations—iminium ions—and alkyl radicals. Mass spectra of aromatic and cyclic aliphatic amines exhibit strong molecular ion peaks, but acyclic aliphatic amines show...
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
Mass Spectrometry: Amine Fragmentation00:55

Mass Spectrometry: Amine Fragmentation

Amines can be identified using mass spectroscopy based on their characteristic fragmentation patterns. The molecular ions of amines undergo fragmentation via ⍺-cleavage. The ⍺-cleavage of the carbon-carbon bonds in amines generates an alkyl radical and resonance-stabilized nitrogen-containing cation.
In amines, the number of nitrogen atoms affects the mass of the molecular ion, which is described by the nitrogen rule of mass spectrometry. This rule states that a compound containing a single or...
Silica Gel Column Chromatography: Overview01:10

Silica Gel Column Chromatography: Overview

Silica gel column chromatography is a technique for separating compounds using a column packed with silica gel as the stationary phase. This method relies on differences in the polarity of compounds. Based on their polarities, compounds move between the stationary phase (silica gel) and the mobile phase (the solvent), forming discrete bands in the column.
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Affinity Chromatography01:03

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
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Sample Preparation for Analysis: Advanced Techniques

Accurate analysis of complex samples often requires advanced preparation techniques to achieve reliable and reproducible results. Samples containing inorganic or organic materials can be challenging to dissolve or decompose effectively. Standard sample preparation methods include acid digestion, fusion, dry ashing, and wet digestion.
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Biochemical Purification and Proteomic Characterization of Amyloid Fibril Cores from the Brain
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Amino acid analysis using core-shell particle column.

Yanting Song1, Takashi Funatsu, Makoto Tsunoda

  • 1Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|October 2, 2012
PubMed
Summary

Core-shell particle columns offer superior separation efficiency for amino acid analysis compared to traditional columns. A new, rapid High-Performance Liquid Chromatography (HPLC) method using these columns significantly reduces analysis time for amino acids.

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Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Conventional particle-packed and monolithic silica columns have limitations in separation efficiency and speed for amino acid analysis.
  • Core-shell particle technology offers potential advantages in chromatographic performance.

Purpose of the Study:

  • To compare the separation efficiency of core-shell particle columns with conventional chromatography columns.
  • To develop a fast High-Performance Liquid Chromatography (HPLC) method for amino acid determination using a core-shell column.

Main Methods:

  • Comparative analysis of separation efficiency using core-shell, particle-packed, and monolithic silica columns.
  • Development of an HPLC method employing a core-shell Kinetex C18 column.
  • Fluorescence derivatization of amino acids using 4-Fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F).

Main Results:

  • Core-shell columns exhibited smaller theoretical plate heights and maintained separation efficiency at higher flow rates.
  • A rapid HPLC method achieved analysis of 21 NBD-amino acids within 7 minutes.
  • Excellent linearity (40 fmol to 40 pmol) and accuracy (90.9-107%) were demonstrated for amino acid quantification.

Conclusions:

  • Core-shell particle columns provide enhanced separation efficiency and speed for amino acid analysis.
  • The developed HPLC method is significantly faster than previous approaches using conventional columns.
  • This method shows promise for rapid amino acid determination in biological samples.