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

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.