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Related Concept Videos

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,...
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...
High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
Optimizing Chromatographic Separations01:15

Optimizing Chromatographic Separations

Optimizing chromatographic separations is crucial for obtaining clean separations in a minimum amount of time. Optimization is required for several factors, including kinetic effects related to band broadening, plate height, capacity factor, and separation factor.
Band broadening refers to spreading solute bands as they travel through the column. This broadening can impact resolution. Plate height (H) represents the length required for one theoretical plate. A lower plate height corresponds to...
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:
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: Jul 15, 2026

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry
09:34

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry

Published on: September 20, 2016

Fast size-exclusion chromatography at high temperature.

Soojin Park1, Heesook Cho, Youngtak Kim

  • 1Department of Chemistry and Polymer Research Institute, Pohang University of Science and Technology, Pohang 790-784, South Korea.

Journal of Chromatography. A
|May 8, 2007
PubMed
Summary

High-temperature size-exclusion chromatography (SEC) significantly reduces analysis time. This method uses high column temperatures and backpressure to prevent eluent boiling, maintaining resolution for faster, efficient SEC analysis.

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Last Updated: Jul 15, 2026

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry
09:34

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry

Published on: September 20, 2016

Simple In-House Ultra-High Performance Capillary Column Manufacturing with the FlashPack Approach
13:36

Simple In-House Ultra-High Performance Capillary Column Manufacturing with the FlashPack Approach

Published on: December 4, 2021

Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline
11:09

Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline

Published on: January 5, 2017

Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Size-exclusion chromatography (SEC) is a widely used technique for polymer and protein analysis.
  • Traditional SEC methods can be time-consuming, limiting throughput.

Purpose of the Study:

  • To investigate the feasibility of operating size-exclusion chromatography (SEC) at high column temperatures.
  • To reduce overall analysis time without compromising separation resolution.

Main Methods:

  • High column temperature operation was employed, exceeding the eluent's boiling point.
  • Sufficient column backpressure was applied to maintain the mobile phase in a liquid state.
  • Narrow bore tubing was inserted between the column and detector to cool the effluent and prevent detector damage.

Main Results:

  • Significant reduction in SEC analysis time was achieved.
  • High-temperature operation did not lead to a serious deterioration in separation resolution.
  • Standard SEC detectors were compatible with the modified setup without modification.

Conclusions:

  • High-temperature SEC is a viable method for accelerating chromatographic analyses.
  • This approach offers a practical solution for increasing the efficiency of SEC workflows.
  • The method allows for faster characterization of macromolecules using conventional instrumentation.