Innovation in pharmaceutical industry in the current decade, both in therapeutics formulations and diagnostic kits, is mainly driven by large biomolecules as active substances. Whether such molecules belong to the class of proteins or nucleotides, they are inherently complex and require substantial analysis and characterization effort, both in development as well as in manufacturing, where e.g., an extended list of critical quality attributes must be monitored for their release control. With HPLC and LC-MS being the predominant analytical techniques, analysis throughput is generally limited and a speed-up of separation methods with UHPLC technology can reduce the critical time from sample to result only to a limited extent.
An elegant way to compensate for the runtime limitation is to use multiple flow paths at the time, which allows to run anything from a single injection to two injections fully in parallel. The highly versatile technology of the Thermo Scientific Vanquish Duo (U)HPLC systems supports this concept very effectively by their dual-flow-path setup allowing the overlapping of two independent injections. The initial design driver was the evolution of 2D-LC that strove for increased peak capacities and the benefit of orthogonal selectivities in different separation dimensions. Such instrumentation, though, can also facilitate the automated combination of different steps in a workflow and enable throughput increase without changing established separation methods. An additional benefit lies in maximizing the data output of high-end mass spectrometers by minimizing idle times, thus boosting laboratory productivity. This presentation will elucidate different ways of throughput increase by executing separation methods partially or completely simultaneously. Examples from the space of 1D-LC-MS will address tandem operation of gradient methods, interlacing of two methods to have the MS continuously generating relevant data, all the way to doubling throughput with a 2-channel set-up for dual optical detection. From the space of 2D-LC, the simultaneous determination of critical attributes in different chromatography dimensions will also be highlighted.