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Column Efficiency: Rate Theory01:12

Column Efficiency: Rate Theory

The rate theory of chromatography provides quantitative insight into the shapes and widths of elution bands. These bands are based on the random-walk mechanism governing molecular migration within a column. The Gaussian profile of chromatographic bands arises from the cumulative effect of random molecular motions as they progress through the column.
During elution, a solute molecule experiences numerous transitions between stationary and mobile phases, exhibiting irregular residence times in...
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...
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.
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Extraction: Advanced Methods00:56

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Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is formed in...
High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

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Supercritical Fluid Chromatography01:18

Supercritical Fluid Chromatography

Supercritical fluid chromatography (SFC) provides a beneficial substitute for gas chromatography (GC) and liquid chromatography (LC) for certain samples because it merges the top attributes of both techniques. SFC allows the separation and analysis of compounds that GC or LC does not easily manage. These compounds are traditionally nonvolatile or thermally unstable, making GC unsuitable and lacking functional groups required for HPLC analysis.
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Related Experiment Video

Updated: Jul 4, 2026

Fabricating High-viscosity Droplets using Microfluidic Capillary Device with Phase-inversion Co-flow Structure
08:02

Fabricating High-viscosity Droplets using Microfluidic Capillary Device with Phase-inversion Co-flow Structure

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Hydrodynamics and mass transfer in two-phase aqueous extraction using spray columns.

S B Sawant1, S K Sikdar, J B Joshi

  • 1Center for Chemical Technology, National Institute of Science and Technology, Boulder, CO 80303, USA.

Biotechnology and Bioengineering
|June 20, 1990
PubMed
Summary

This study shows polyethylene glycol (PEG) and dextran two-phase aqueous extraction in spray columns effectively purifies proteins. Protein mass transfer is independent of PEG velocity, indicating efficient purification.

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Area of Science:

  • Biochemical Engineering
  • Separation Science
  • Protein Purification

Background:

  • Two-phase aqueous extraction using polyethylene glycol (PEG) and dextran is a viable method for protein isolation.
  • Spray columns offer a continuous processing approach for such extractions.

Purpose of the Study:

  • To investigate the impact of PEG phase velocity on dispersed phase holdup and mass transfer coefficients in a spray column.
  • To determine the efficiency of protein purification using this technique under varying flow conditions.

Main Methods:

  • Experiments were conducted in a 9.7 mm internal diameter spray column.
  • Fractional dispersed phase (PEG) holdup was measured.
  • Overall mass transfer coefficients for bovine serum albumin were determined.

Main Results:

  • Dispersed phase holdup increased with increasing PEG phase velocity.
  • Overall mass transfer coefficients for bovine serum albumin, when normalized for PEG holdup, were independent of PEG phase velocity.

Conclusions:

  • The purification efficiency of bovine serum albumin is not limited by phase velocity in this spray column system.
  • Spray column extraction with PEG-dextran systems provides a robust method for protein purification, with mass transfer independent of flow rates.