Development of a Novel Chromatographic Method for Analysis of Drugs and Formulations

The pharmaceutical industry requires robust, validated analytical methods for accurate drug quantification and quality control. This study aimed to develop and validate novel stability-indicating chromatographic methods for three therapeutically diverse drugs: Benzgalantamine (Alzheimer's disease), Ensifentrine (COPD), and Tovorafenib (oncology) using advanced chromatographic techniques integrated with sustainable analytical practices. A comprehensive analytical approach was employed utilizing high-
performance liquid chromatography (HPLC), ultra-performance liquid chromatography (UPLC), and liquid chromatography-mass spectrometry (LC-MS). The methodology incorporated Analytical Quality by Design (AQbD) principles and green chemistry concepts to optimize analytical performance while minimizing environmental impact. Method development followed ICH Q2(R1) validation guidelines, including assessment of accuracy, precision, specificity, linearity, detection limits, and robustness. Stability-indicating capacity was established through forced degradation studies under varied stress conditions (acid/base hydrolysis, oxidative degradation, photolysis, and thermal stress) as per ICH Q1A(R2) guidelines. The developed chromatographic methods demonstrated exceptional analytical performance across all three drug
compounds. HPLC methods successfully achieved baseline separation of parent drugs from their degradation products, confirming stability-indicating capability. UPLC methodology reduced analysis time by approximately 50% compared to conventional HPLC while maintaining superior resolution and significantly
decreasing solvent consumption, aligning with green analytical chemistry principles. LC-MS methods provided enhanced sensitivity and selectivity, enabling simultaneous multi-analyte detection with improved specificity for complex formulation analysis. The systematic AQbD approach identified critical method parameters including pH optimization, column temperature control, and mobile phase composition. Design
of Experiments (DoE) statistical modeling established optimal parameter ranges, while Method Operable Design Region (MODR) definition ensured robust analytical performance and reproducibility across different laboratory conditions. This research successfully established validated, stability-indicating
chromatographic methods for Benzgalantamine, Ensifentrine, and Tovorafenib analysis. The integration of
advanced chromatographic techniques with QbD principles and green chemistry considerations resulted in
robust, environmentally sustainable analytical methodologies suitable for routine pharmaceutical quality control, stability testing, and regulatory compliance. These methods address a significant analytical gap in the literature while providing a framework for sustainable pharmaceutical analysis.