Ultra high performance liquid chromatography (UHPLC) provides improved separation speed, throughput, and sensitivity by employing stationary phase particles of around 2 μm or smaller. UHPLC has found widespread use in the analysis of small molecules in pharmaceutical, food, and environmental areas. Are these analytical benefits also applicable to the separation of larger molecules, such as proteins and peptides? Biomolecules have, apart from their size, other differences from small molecules (e.g. charges, complex structures) that make the application of UHPLC not as straightforward as for small molecules. This article discusses the rationale of using UHPLC in life sciences and demonstrate where the potential is. It also discusses the requirements needed for a true bio UHPLC system and shows examples of relevant biomolecule separations.