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Ultra-high performance supercritical fluid chromatography in impurity control II: Method validation.

Kateřina Plachká1, Maria Khalikova1, Barbora Babičová1

  • 1Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Heyrovského 1203, 500 05, Hradec Králové, Czech Republic.

Analytica Chimica Acta
|May 16, 2020
PubMed
Summary

Ultra-high performance supercritical fluid chromatography (UPS2) methods were optimized and validated for pharmaceutical impurity control. Most methods achieved required resolution and sensitivity for drug substance and product analysis.

Keywords:
Impurity controlMethod validationPharmaceutical compoundsQuality controlUltra-high performance supercritical fluid chromatography

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Area of Science:

  • Analytical Chemistry
  • Pharmaceutical Analysis

Background:

  • Ultra-high performance supercritical fluid chromatography (UPS2) offers a promising approach for pharmaceutical quality control.
  • Previous work established a screening method for 10 pharmaceutical mixtures using UPS2, with most achieving baseline separation.

Purpose of the Study:

  • To optimize and validate UPS2 methods for pharmaceutical impurity control, addressing challenges in resolution, retention time stability, and method applicability.
  • To ensure methods meet stringent validation criteria for impurity detection in drug substances and products.

Main Methods:

  • Optimization of gradient programs, column selection (Viridis UPC2 HSS C18 SB vs. Acquity UPLC HSS C18 SB), modifier composition (acetonitrile addition), and column coupling.
  • Validation of 10 UPS2 methods according to ICH guidelines (Q2, Q3), assessing specificity, linearity, range, LOQ, LOD, accuracy, and precision.
  • Determination of intermediate precision and accuracy profiles for selected methods.

Main Results:

  • Method adjustments, primarily gradient changes and column substitutions, improved selectivity and resolution.
  • Most methods successfully met validation criteria, demonstrating applicability for impurity control.
  • Two impurities failed validation due to insufficient resolution and sensitivity, with only the identification threshold met.

Conclusions:

  • Optimized UPS2 methods are suitable for pharmaceutical impurity control, offering high separation efficiency.
  • While most impurities were quantifiable, challenges with resolution and sensitivity for specific compounds remain.
  • Further refinement may be needed for certain challenging impurity profiles to meet all validation requirements.