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Related Concept Videos

Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

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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...
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Principles Of Column Chromatography01:13

Principles Of Column Chromatography

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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...
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High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

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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.
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High-Performance Liquid Chromatography: Elution Process01:05

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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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Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
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High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

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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 Videos

pH-zone-refining counter-current chromatography: origin, mechanism, procedure and applications.

Yoichiro Ito1

  • 1Laboratory of Bioseparation Technology, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, 10 Center Drive, Bethesda, MD 20892, USA. itoy2@mail.nih.gov

Journal of Chromatography. A
|December 11, 2012
PubMed
Summary

pH-zone refining counter-current chromatography (CCC) enhances separation by using acids to form sharp peaks. This novel method significantly increases sample loading capacity and improves purification yields for various compounds.

Related Experiment Videos

Area of Science:

  • Analytical Chemistry
  • Separation Science

Background:

  • High-speed counter-current chromatography (HSCCC) has been the standard for natural and synthetic product purification since 1980.
  • A novel elution mode, pH-zone refining CCC, emerged in 1991, utilizing organic acids to create sharp analyte peaks.

Purpose of the Study:

  • To review the principles and applications of pH-zone refining CCC.
  • To highlight its advantages over conventional HSCCC, including increased sample capacity and improved purification efficiency.

Main Methods:

  • Investigated the mechanism of sharp peak formation using bromoacetic acid as a retainer.
  • Developed a new method employing retainers and eluters for controlled border movement.
  • Illustrated the hydrodynamics with acidic samples and presented typical separations, including affinity and chiral separations.

Main Results:

  • pH-zone refining CCC demonstrated a tenfold increase in sample loading capacity compared to conventional HSCCC.
  • Fractions were highly concentrated, yields improved with larger sample sizes, and charged compounds were concentrated at peak boundaries.
  • Minute charged compounds and compounds lacking chromophores were effectively concentrated and detected.

Conclusions:

  • pH-zone refining CCC offers significant advantages for separation and purification, including higher capacity, better yields, and enhanced detection capabilities.
  • The method has seen increasing research interest since 1994, with over 70 papers published.