Peak Purity of Peptides and Large Molecules by Accurate Mass LC-MS

Purity of a chromatographic peak has traditionally been analysed by HPLC analysis using 3D UV to assess compounds spectrally different to the Active Pharmaceutical Ingredient (API) peak. This is a well-established methodology that is routinely and widely used to assess peak purity of small molecules, relying on differences in the 3D UV spectra to capture potential co-eluting impurities.

In this paper, two example case studies are presented for peak purity determination. Firstly, for a typical peptide, determined using Agilent 6230/6530 Accurate Mass TOF/QTOF instrumentation and Agilent MassHunter B.10 software. Secondly, a larger biologics product in the form of a monoclonal antibody determined using a Thermo Q-Exactive Plus BioPharma Orbitrap MS with Chromeleon 7.2.10 Software and BioPharma Finder 4.3 Software.