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Related Concept Videos

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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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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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The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
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A workflow for column interchangeability in liquid chromatography using modeling software and quality-by-design

Róbert Kormány1, Katalin Tamás1, Davy Guillarme2

  • 1Egis Pharmaceuticals Plc., Keresztúri út 30-38, Budapest 1106, Hungary.

Journal of Pharmaceutical and Biomedical Analysis
|September 9, 2017
PubMed
Summary

This study presents a workflow for evaluating chromatographic resolution using LC modeling software. It enables easy identification of interchangeable columns, ensuring method robustness and reliable impurity analysis.

Keywords:
Column interchangeabilityDesign of experimentsDryLabMethod developmentQuality by designUHPLC

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Area of Science:

  • Analytical Chemistry
  • Chromatography
  • Method Development

Background:

  • Optimizing chromatographic resolution is crucial for accurate separation of analytes and impurities.
  • Traditional method development can be time-consuming and resource-intensive.
  • Identifying column interchangeability is essential for robust analytical methods.

Purpose of the Study:

  • To develop a generic workflow for evaluating chromatographic resolution across a wide design space.
  • To facilitate the identification of suitable replacement columns for existing methods.
  • To ensure method robustness and reliability through systematic column evaluation.

Main Methods:

  • Utilized modern Liquid Chromatography (LC) modeling software (Drylab) for predicting chromatographic behavior.
  • Employed a limited experimental design approach (2-3 hours per column) for data acquisition.
  • Modeled resolution across the entire design space using sub-2μm particle columns (UHPLC).
  • Overlapped design spaces from different columns to identify robust operating regions.

Main Results:

  • Successfully modeled chromatographic resolution in a large design space with minimal experimental effort.
  • Demonstrated the interchangeability of columns from different manufacturers (Waters Acquity HSS C18, Thermo Hypersil Gold C18).
  • Identified a working point providing sufficient resolution and robustness for amlodipine and its impurities.

Conclusions:

  • The developed workflow provides an efficient and reliable method for chromatographic column selection and method transfer.
  • LC modeling software significantly accelerates the evaluation of chromatographic resolution and method robustness.
  • The case study confirmed the practical applicability of the workflow for pharmaceutical impurity analysis.