Theoretically, if a 3:1 solvent:sample ratio is used for protein precipitation during the filtration steps, then the final filtrate will be approximately 75% acetonitrile. If the initial liquid chromatography mobile phase composition is not close to 75% organic solvent, or if poor chromatographic peak shapes are observed for early eluting compounds, we recommend diluting the filtrate in the collection plate with water or aqueous mobile phase to more closely match the initial mobile phase composition. If further dilution is not suitable for your application due to limited instrument sensitivity, evaporation followed by reconstitution with your initial mobile phase can be performed. This approach can be used to obtain a suitable concentration while avoiding peak shape issues that can be caused by unmatched sample solvent and initial mobile phase.
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