Nico C. van de Merbel

The technical requirements for a successful LC–MS/MS method for the quantitation of biopharmaceuticals are presented and the advantages and disadvantages compared to ligand-binding assays are evaluated.

Advances in Liquid Chromatography–Tandem Mass Spectrometry (LC–MS–MS)-Based Quantitation of Biopharmaceuticals in Biological Samples

This article describes and compares a number of approaches to increase the speed of liquid chromatographic separations. On a standard LC column, a gain of a factor two in the run time (from 10 to 5 minutes) was achieved by increasing the flow-rate two-fold. On a monolithic column, a column operated at high temperature (120 ?C) and a short column, flow-rates could be increased typically five-fold, resulting in run times in the order of 2 minutes. This was accompanied by a sometimes considerable loss in separation efficiency. A combination of a very short run time and unaffected separation efficiency was realized on a UPLC system, designed for use at higher pressure. By working at approximately 800 bar, the analytes could be well separated within 30 seconds.