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Related Concept Videos

Gas Chromatography: Types of Columns and Stationary Phases01:17

Gas Chromatography: Types of Columns and Stationary Phases

Gas chromatography (GC) relies on stationary phases to separate and analyze components in a sample. There are two main types of stationary phases: liquid and solid. Liquid stationary phases are non-volatile, thermally stable, and chemically inert liquids coated onto the column. Solid stationary phases are particles of adsorbent material, such as silica gel or molecular sieves.
For an analyte to remain on the column for a sufficient amount of time, it must exhibit some level of compatibility (or...
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...
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...
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...
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...

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Related Experiment Video

Updated: Jun 4, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
06:25

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

Published on: April 26, 2016

Preparation of reversed-phase microcolumns.

Martin R Larsen

    CSH Protocols
    |March 2, 2011
    PubMed
    Summary

    This study details fabricating reversed-phase microcolumns from GELoader tips for sample cleanup before MALDI-MS analysis. This method also works for Electrospray Ionization-Mass Spectrometry (ESI-MS) with minor adjustments.

    Area of Science:

    • Analytical Chemistry
    • Mass Spectrometry

    Background:

    • Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry (MALDI-MS) requires effective sample cleanup.
    • Microscale columns offer a versatile approach for direct sample elution onto MALDI targets.

    Purpose of the Study:

    • To describe the fabrication of a reversed-phase microcolumn for sample cleanup.
    • To optimize the microcolumn for MALDI-MS analysis.
    • To adapt the method for Electrospray Ionization-Mass Spectrometry (ESI-MS).

    Main Methods:

    • Fabrication of microcolumns using GELoader tips.
    • Development of a reversed-phase chromatography protocol.
    • Optimization for direct elution onto MALDI target plates.

    Main Results:

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    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

    Related Experiment Videos

    Last Updated: Jun 4, 2026

    Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
    06:25

    Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

    Published on: April 26, 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

    • Successful fabrication of functional reversed-phase microcolumns.
    • Demonstrated utility for sample cleanup prior to MALDI-MS.
    • Verified applicability for ESI-MS with modifications.

    Conclusions:

    • The described microcolumn fabrication is an effective method for sample cleanup in mass spectrometry.
    • This technique is adaptable for both MALDI-MS and ESI-MS workflows.