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Protein separations are sensitive to pH and ionic strength because these solution characteristics directly affect the structure of the molecule and its interaction with the packing material.While screening combinations of these parameters is recognized as one of the best paths to an optimized chromatographic method, the preparation of a series of buffers for experimental evaluation is time-consuming and error prone.A protocol has been developed for using the chromatographic system to prepare the required combinations of buffer on demand so that separation space can be systematically explored.A set of algorithms allows a user interface where the chromatographic method can be programmed directly in units of pH and salt concentration.This is known as Auto·Blend PlusTM Technology, and is implemented on a four-solvent ACQUITY UPLC SystemTM.