A single LC‑MS injection is usually not sufficient for HCP quantification because the workflow includes multiple sources of variability, such as sample preparation, chromatography, and instrument performance. In addition, data acquisition can introduce stochastic effects, meaning low‑abundance HCPs may be missed or inconsistently quantified in a single run.

Including replicate injections helps assess instrument precision and improves confidence in both identification and quantification. Where higher robustness is required, analysing independent sample replicates also captures variability from sample handling and digestion. Together, this approach provides more reliable and GMP‑relevant data.

Thomas Kofoed, PhD, CEO of Alphalyse in Denmark, discussed key factors in implementing GMP-compatible LC-MS methods for release assays and highlighted successful cases where this approach has replaced traditional ELISA methods. Don’t miss his presentation and expert insights at the panel discussion.