Several factors contribute to the generation of high quality and reproducible UPLC-based, protein SEC separations. The selection of an appropriately configured Waters UPLC Instrument and the proper attachment of the SEC column to the LC system are major factors contributing to the generation of successful chromatography.
Table 1 includes the recommended Waters UPLC System components for use in SEC separations that are frequently performed using salt-containing mobile phases that lack organic solvent. As noted in Figure 2, the use of non-recommended ACQUITY UPLC System components can have a significantly negative effect on the quality of the SEC separation. (For more information, reference: "Guidelines for Routine Use and Maintenance of Ultra-Performance Size-Exclusion and Ion-Exchange Chromatography Systems.")
Figure 2. ACQUITY UPLC Protein BEH SEC 200Å chromatogram of bovine serum albumin (BSA) using an ACQUITY UPLC Photodiode Array Detector that shows the adverse effect of the flow-cell material of construction on peak shape. The BSA monomer exhibits extensive peak tailing when the flow cell consists of Teflon AF material.
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