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Pharmaceutical impurity analysis by comprehensive two-dimensional temperature responsive × reversed phase liquid

Kristina Wicht1, Mathijs Baert1, Ardiana Kajtazi1

  • 1Separation Science Group, Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281-S4, B-9000 Ghent, Belgium.

Journal of Chromatography. A
|September 29, 2020
PubMed
Summary

Temperature-responsive × reversed-phase liquid chromatography (TRLC × RPLC) offers a novel, optimization-free approach for pharmaceutical impurity analysis. This method enhances peak capacity, overcoming limitations of traditional techniques for complex drug formulations.

Keywords:
Comprehensive two-dimensional liquid chromatographyImpurity determinationPharmaceutical analysisRefocusingTemperature responsive liquid chromatography

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Area of Science:

  • Analytical Chemistry
  • Chromatography Science

Background:

  • Two-dimensional Liquid Chromatography (2D-LC) is increasingly relevant for analyzing complex pharmaceutical formulations due to higher peak capacity.
  • One-dimensional High-Performance Liquid Chromatography (1D-HPLC) often proves insufficient for detecting the numerous impurities in such formulations.
  • Existing 2D-LC methods face challenges including modulation, solvent compatibility, orthogonality, and sensitivity.

Purpose of the Study:

  • To assess the potential of Temperature-Responsive × Reversed-Phase Liquid Chromatography (TRLC × RPLC) for pharmaceutical impurity analysis.
  • To explore a generic 2D-LC approach that minimizes method development time and effort.
  • To overcome common limitations associated with current 2D-LC techniques.

Main Methods:

  • Investigated TRLC × RPLC using a mixture of 11 corticosteroids and 6 progestogens.
  • Evaluated orthogonality and peak capacities across three different Reversed-Phase (RP) core-shell column selectivities (Poroshell EC-C18, phenyl-hexyl, PFP).
  • Employed full, shifted, or segmented gradients for separation optimization based on mixture composition.

Main Results:

  • The TRLC × PFP column combination demonstrated superior performance for the specific separation.
  • The TRLC × RPLC platform enabled impurity detection at the 0.05% level relative to a main compound.
  • Systematic and optimization-free refocusing in the second dimension was achieved by combining aqueous TRLC with RPLC.

Conclusions:

  • TRLC × RPLC presents a promising, low-development strategy for pharmaceutical impurity profiling.
  • This approach is suitable for analyzing complex formulations, including those with multiple active ingredients.
  • The method overcomes significant hurdles in generic 2D-LC, offering enhanced analytical capabilities.