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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:
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...
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...
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...
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...

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Related Experiment Video

Updated: Jul 13, 2026

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
10:17

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

Published on: April 23, 2019

Efficient method development strategy for challenging separation of pharmaceutical molecules using advanced

Kang Ping Xiao1, Yuan Xiong, Fang Zhu Liu

  • 1Global Quality Services - Analytical Sciences, Schering-Plough Corporation, Union, NJ 07083, USA.

Journal of Chromatography. A
|July 14, 2007
PubMed
Summary

This study presents an efficient strategy for separating challenging pharmaceutical molecules using advanced chromatography tools, significantly saving time for scientists. The method successfully resolved difficult-to-separate epimers with high resolution.

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Automated HPLC Separation Using LC-Mate: An Integrated Repetitive Autosampler and Fraction Collector for Microscale Purification
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Automated HPLC Separation Using LC-Mate: An Integrated Repetitive Autosampler and Fraction Collector for Microscale Purification

Published on: February 27, 2026

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Last Updated: Jul 13, 2026

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
10:17

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

Published on: April 23, 2019

Automated HPLC Separation Using LC-Mate: An Integrated Repetitive Autosampler and Fraction Collector for Microscale Purification
07:11

Automated HPLC Separation Using LC-Mate: An Integrated Repetitive Autosampler and Fraction Collector for Microscale Purification

Published on: February 27, 2026

Area of Science:

  • Analytical Chemistry
  • Pharmaceutical Sciences

Background:

  • Developing high-performance liquid chromatography (HPLC) methods for complex pharmaceutical molecules can be time-consuming.
  • Structural isomers like epimers often present significant separation challenges in pharmaceutical analysis.

Purpose of the Study:

  • To describe an efficient strategy for developing HPLC separations of challenging pharmaceutical molecules.
  • To demonstrate the utility of advanced chromatographic technologies for method development.

Main Methods:

  • Utilized a computer-assisted method development tool (ChromSword) for mobile phase screening and optimization.
  • Employed an automated column switching system (LC Spiderling) for HPLC column selection.
  • Applied the strategy to separate structural epimers: betamethylepoxide and alphamethylepoxide.

Main Results:

  • Achieved a rapid separation of betamethylepoxide and alphamethylepoxide in 10 minutes with a resolution factor of 3.0.
  • Demonstrated baseline separation even with a 10,000:1 concentration ratio of the epimers.
  • The developed strategy significantly enhanced the probability of achieving adequate separations.

Conclusions:

  • The described strategy, integrating advanced chromatographic tools, offers an efficient approach to developing HPLC methods for challenging pharmaceutical separations.
  • This method development strategy is a significant time-saver for analytical scientists and can be extended to stability-indicating and impurity profiling methods.