Development of a Robust and Repeatable UPLC−MS Method for the Long-Term Metabolomic Study of Human Serum

A method for performing untargeted metabolomic analysis of human serum has been developed based on protein precipitation followed by Ultra Performance Liquid Chromatography and Time-of-Flight mass spectrometry (UPLC−TOF-MS). This method was specifically designed to fulfill the requirements of a long-term metabolomic study, spanning more than 3 years, and it was subsequently thoroughly evaluated for robustness and repeatability. We describe here the observed drift in instrumental performance over time and its improvement with adjustment of the length of analytical block. The optimal setup for our purpose was further validated against a set of serum samples from 30 healthy individuals. We also assessed the reproducibility of chromatographic columns with the same chemistry of stationary phase from the same manufacturer but from different production batches. The results have allowed the authors to prepare SOPs for “fit for purpose” long-term UPLC−MS metabolomic studies, such as are being employed in the HUSERMET project. This method allows the acquisition of data and subsequent comparison of data collected across many months or years.