In-column dilution: Improving volume loadability in reverse phase gradient chromatography through the use of a silica pre-column.

•Use of silica pre-column significantly reduces impact of large volume injections in preparative reversed phase gradient purifications.•In-Column Dilution approach allows use of smaller columns for preparative purifications, resulting in significant solvent and cost reduction.•Injection volumes can be increased by a factor of three when using appropriately sized silica pre-columns.•In-Column Dilution can also benefit analytical HLC analyses that require large volume injections.