Trapping mode two-dimensional liquid chromatography for quantitative low-level impurity enrichment in pharmaceutical...

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis
• Chromatography

Background:

• Two-dimensional liquid chromatography (2D-LC) is used in pharmaceutical analysis for analyte enrichment.
• Multi-trapping 2D-LC shows potential for low-level impurity monitoring, but its quantitative aspects are not well-established.
• Existing methods struggle with impurity levels from parts-per-million (ppm) to 0.15% (w/w).

Purpose of the Study:

• To evaluate the quantitative capabilities of a multi-trapping 2D-LC workflow for low-level pharmaceutical impurity analysis.
• To demonstrate the application of this system in real-world pharmaceutical impurity case studies.
• To establish a robust, accessible method for impurity monitoring in quality control.

Main Methods:

• Development of a simple heart-cutting trapping mode 2D-LC workflow using standard 1D-LC components.
• Evaluation of quantitative performance using standard markers, assessing linear enrichment and recovery.
• Application of the developed system to identify and quantify impurities in pharmaceutical samples.

Main Results:

• Demonstrated linear enrichment for up to 20 trapping cycles with over 97.0% recovery.
• Successfully identified unknown impurities at sub-ppm levels and a new impurity at 0.05% (w/w).
• Quantified a potential mutagenic impurity at 10-ppm level with >97.0% recovery and <3.0% RSD.

Conclusions:

• The developed multi-trapping 2D-LC workflow is robust, accurate, and precise for low-level impurity monitoring.
• The system requires no specialized equipment, making it suitable for quality control laboratories.
• This method enables the development of validated low-impurity monitoring methods for pharmaceutical analysis.