Compounds should be dissolved in the mobile phase for best results during purification. Using mobile phase as the dissolution solvent avoids disturbing equilibration of the column and maximizes resolution between peaks. Unfortunately, complex mixtures contain compounds that may not be soluble in the mobile phase. Dimethyl sulfoxide (DMSO) and dimethylforamide (DMF) are often used in reverse phase chromatography because they dissolve a wide variety of compounds and are “weaker” solvents compared to the “strong” solvent in the mobile phase, which causes the compounds to elute. These solvents are still stronger eluting solvents than water, causing limited sample loading. On-column dilution is a method that allows the use of DMSO and DMF while preserving peak shape and loading capacity. The use of On-Column Dilution allows 2 to 3 fold improvement in purification throughput.

On-column dilution: A method to improve loading and resolution in chromatography

2019 Spring National Meeting

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