Stability-Indicating RP-UPLC Analysis of Semaglutide: Integrating Green Analytical Chemistry Principles in Bulk and Pharmaceutical Formulations

Abstract

Aim and Objectives

The aim of this study was to develop and validate a simple, precise, sensitive and stability-indicating method for the separation and determination of semaglutide in pharmaceutical formulations, in compliance with ICH Q2(R1) guidelines and Green Analytical Chemistry principles.

Methods

Chromatographic separation was achieved using a Phenyl XBD column (100 × 2.6 mm, 1.7 μm) with a mobile phase of 0.1% formic acid and acetonitrile (70:30 v/v) at a flow rate of 0.5 mL/min. Detection was performed at 294 nm, and retention time (RT) of semaglutide was observed at 1.729 min. Linearity was evaluated across 2–12 µg/mL, precision was assessed using %RSD, and sensitivity was determined through LOD and LOQ values.

Results

The method showed excellent linearity with R² = 0.999. System and method precision were confirmed with %RSD values ranging from 0.35 to 0.72. Recovery studies demonstrated values between 100.06% and 100.6%. The method exhibited high sensitivity with LOD of 0.083 µg/mL and LOQ of 0.61 µg/mL. Stress degradation studies confirmed complete separation of degradants formed under acid, alkali hydrolysis and oxidation, with good resolution, establishing the method’s stability-indicating nature.

Conclusion

The validated method is precise, accurate, and robust for semaglutide analysis under varied stress conditions. Importantly, it aligns with Green Analytical Chemistry principles by minimizing solvent use, employing a safer aqueous-organic mobile phase, and reducing environmental impact without compromising analytical performance.