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Development of a Fast and Robust UHPLC Method for Apixaban In-Process Control Analysis.

Róbert Kormány1, Norbert Rácz1, Szabolcs Fekete2

  • 1Drug Substance Analytical Development Division, Egis Pharmaceuticals Plc., Keresztúri út 30-38, H-1106 Budapest, Hungary.

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|July 2, 2021
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Summary

Quality by Design (QbD) optimizes high-performance liquid chromatography (HPLC) for pharmaceutical in-process control. This study validates software for rapid UHPLC method development, ensuring robust analysis of synthesis mixtures like apixaban.

Keywords:
apixabandesign of experimentsliquid chromatographymethod developmentquality by designrobustness

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Area of Science:

  • Analytical Chemistry
  • Pharmaceutical Analysis
  • Chromatography

Background:

  • In-process control (IPC) is crucial in pharmaceutical synthesis.
  • High-performance liquid chromatography (HPLC) is the standard IPC technique.
  • Quality by Design (QbD) principles enhance HPLC method development over traditional approaches.

Purpose of the Study:

  • To evaluate multifactorial liquid chromatographic optimization software within an expanded knowledge space.
  • To assess the application of ultra-high performance liquid chromatography (UHPLC) for rapid IPC analysis.
  • To determine a robust working range for method parameters in apixaban synthesis analysis.

Main Methods:

  • Utilized ultra-high performance liquid chromatography (UHPLC) with a short, narrow-bore column (50 × 2.1 mm, sub-2 µm particles).
  • Employed a multifactorial liquid chromatographic optimization software for method development.
  • Investigated a narrow range of method parameters: gradient time (tG), temperature (T), and mobile phase pH.
  • Compared virtual robustness testing results with experimental measurements and Design of Experiments (DoE) predictions.

Main Results:

  • UHPLC enabled significant reduction in analysis time, with reaction mixture composition analyzed within minutes.
  • The optimization software effectively predicted chromatographic conditions.
  • Virtual robustness testing results showed good agreement with experimental data and DoE predictions.
  • A robust working zone for the apixaban synthesis mixture analysis was successfully identified.

Conclusions:

  • Multifactorial optimization software is applicable for UHPLC method development in pharmaceutical IPC.
  • QbD principles combined with UHPLC and optimization software facilitate rapid and robust analytical method development.
  • This approach supports efficient and reliable in-process control during pharmaceutical manufacturing.