Size Exclusion Chromatography Method Optimization using the European Pharmacopoeia for Insulin Fibrils

Insulin is a peptide hormone which plays a central role in metabolism. In its monomer form, insulin can partially misfold, causing a cascade of non-native cross assembly of beta-pleated sheets or amyloid-like fibrils. A common method for analyzing insulin HMWP is size exclusion chromatography (SEC). However, the method requires a lengthy 35 minute run time which is time prohibitive for any laboratory with many samples to run and data points to generate. This application shows the method transfer of the Ph. Eur. method from a traditional, 10µm hydrophilic silica 300 x 7.8mm column to a high performance, 1.8µm hydrophilic silica 150 x 4.6mm column.