Green and High-Throughput Quantification of Semaglutide in Bulk Substance and Solutions for Injections Using Liquid Chromatography

Abstract

A rapid and robust reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and fully validated strictly as a quantitative assay for the high-throughput determination of semaglutide (SMG) in both active pharmaceutical ingredient (API) powder and solutions for injection. Chromatographic separation was achieved on a Phenomenex Aeris™ 3.6 µm Widepore XB-C18 column (150 × 2.1 mm) using a mobile phase consisting of 100 mM ammonium dihydrogen phosphate buffer combined with acetonitrile , with the addition of perchloric acid using an isocratic elution. The advantage of the developed methodology is the compression of the total run time to 6 minutes, which introduces an over three-fold increase in throughput compared to existing methods. The method was successfully validated according to ICH Q2(R2) , showing specificity, linearity (R2 = 0.998 within 70%–130% range), precision (%RSD ≤ 1.73%) and accurate recovery (98.8% – 100.2%) for bulk and final formulation samples.