Abstract

Method development for synthetic peptide molecules is often a challenging task as peptides get easily degraded in mild stress conditions and also offer chromatographic challenges like carryover, peptide loss during chromatographic run and adsorption of the drug onto the walls of the storage container. This article presents an easy, accurate and reproducible stability-indicating, mass compatible Reverse Phase-Liquid Chromatographic method for the detection of liraglutide in bulk active pharmaceutical ingredient. The stress conditions chosen for the study (hydrolytic, oxidative, photolytic and thermal) were as per the International Council for Harmonization guideline. Chromatographic separation of liraglutide from its degradation products was achieved on a bioZen™ 2.6 μm Peptide XB-C18 100 Å, liquid chromatography column 150×4.6 mm, using ammonium formate buffer (pH 3.0) and acetonitrile (0.1 % formic acid) as the mobile phase in gradient elution mode. The elution was carried out at a flow rate of 0.7 ml/min. Detection of liraglutide and its degradants were monitored at 215 nm. Liraglutide was found to degrade significantly in hydrolytic, oxidative and thermal stress conditions. Apart from stress testing, adsorption study for liraglutide was performed using 5 different types of containers. Also, several buffers/diluents were screened to reduce the carryover of active pharmaceutical ingredient in subsequent chromatographic runs.