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Related Concept Videos

High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:
Principles Of Column Chromatography01:13

Principles Of Column Chromatography

The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...
Types Of Column Chromatography01:29

Types Of Column Chromatography

The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
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.
Polar components tend to bind strongly to the silica gel, causing them to move slowly through the column. In contrast, nonpolar compounds...
Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...
High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

High-performance liquid chromatography, or HPLC, is an analytical technique that separates liquid samples under high pressures. An HPLC instrument consists of glass bottles for storing solvents called mobile phase reservoirs. HPLC-grade solvents are used to maintain high purity, and the dissolved gases are removed using a degasser, such as a vacuum pumping system or sparging with helium. The solvents are then pumped into the analytical column using a screw-driven syringe or reciprocating pumps.

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Related Experiment Video

Updated: Jun 14, 2026

Histone Modification Screening using Liquid Chromatography, Trapped Ion Mobility Spectrometry, and Time-Of-Flight Mass Spectrometry
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Histone Modification Screening using Liquid Chromatography, Trapped Ion Mobility Spectrometry, and Time-Of-Flight Mass Spectrometry

Published on: January 12, 2024

Multi-dimensional liquid chromatography in proteomics--a review.

Xiang Zhang1, Aiqin Fang, Catherine P Riley

  • 1Department of Chemistry, University of Louisville, 2320 South Brook Street, Louisville, KY 40292, USA. xiang.zhang@louisville.edu

Analytica Chimica Acta
|April 6, 2010
PubMed
Summary

Proteomics uses mass spectrometry to analyze proteins, but peptide mixtures overwhelm analysis. Multidimensional liquid chromatography (MDLC) enhances peptide separation for better protein identification and quantification in biological systems.

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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)
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Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)

Published on: June 20, 2019

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Last Updated: Jun 14, 2026

Histone Modification Screening using Liquid Chromatography, Trapped Ion Mobility Spectrometry, and Time-Of-Flight Mass Spectrometry
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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)
10:00

Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)

Published on: June 20, 2019

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Proteomics involves the large-scale study of proteins, including their expression, structures, and functions.
  • Current mass spectrometry techniques face limitations in analyzing complex peptide mixtures generated from protein digestion.

Purpose of the Study:

  • To review existing multidimensional liquid chromatographic (MDLC) platforms for proteomics.
  • To discuss the application of MDLC in combination with techniques like stable isotope labeling.
  • To provide perspectives on future developments in proteomic analysis.

Main Methods:

  • Focus on multidimensional liquid chromatography (MDLC) platforms.
  • Integration of MDLC with mass spectrometry for peptide analysis.
  • Application of stable isotope labeling techniques in conjunction with MDLC.

Main Results:

  • MDLC effectively fractionates complex peptide mixtures, overcoming mass spectrometry limitations.
  • Enhanced separation enables improved identification and quantification of peptides.
  • Combined approaches increase the depth and accuracy of proteomic profiling.

Conclusions:

  • MDLC is a crucial technology for advancing proteomics.
  • Optimized chromatographic separations are key to comprehensive protein analysis.
  • Future developments will likely focus on further refining MDLC strategies and integrating novel analytical tools.