A REVIEW OF THE DEVELOPMENT AND VALIDATION OF THE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) METHOD FOR ESTIMATING THE AMINO ACID MOLE RATIO IN SEMAGLUTIDE

Swetha Vegesna, Mishall Masa, Sampath Sri Venkata Sai Kumar, Naga Datta Sasi Mrudula Kothapalli

ABSTRACT

Blood sugar regulation and the treatment of type 2 diabetes depend on semaglutide, an agonist of the glucagon-like peptide-1 (GLP-1) receptor composed of a specific amino acid sequence. A precise determination of the mole ratio and amino acid composition is necessary for a peptide-based treatment to be safe, effective, and of high quality. High-Performance Liquid Chromatography (HPLC) has emerged as the most reliable and versatile analytical technique for peptide and amino acid detection due to its exceptional sensitivity, selectivity, and repeatability. This review focuses on the creation and verification of HPLC methods for measuring amino acid mole ratios in semaglutide. It discusses how important amino acid profiling is for tracking degradation, confirming peptide integrity, and figuring out batch-to-batch consistency. The study emphasizes crucial aspects of method development, including the selection of stationary and mobile phases, optimization of chromatographic conditions, and detection technique selection. The validation standards, which address crucial subjects like specificity, linearity, precision, accuracy, sensitivity, and robustness, are also specified using the International Council for Harmonisation (ICH) guidelines Q2(R1).

Keywords: Semaglutide, HPLC, manufacturing machinery, and the Quality Control division.