Question

In Size Exclusion Chromatography, what can be done if the void volume is > (greater) than 40% of the column???

Answer 1

size-exclusion chromatography is a high-pressure liquid chromatography technique that separates molecules in solution to their size. columns are packed with a porous stationary phase. These columns are used to measure degradation and aggregation of large molecules.

The three parameters Ve,Vo,Vi are used to describe the behavior of a molecule.

Where, Ve – Elution volume

                Vo- Void volume

                 Vi- Inner volume   

The void volume is the volume of liquid. Molecules with diameter larger than pore size are excluded by gel and the elution volume is equal to void volume of the column. The total volume of the column is the sum of void volume and inner volume.

Void volume is >30-40% of the total volume depending on how tightly the column is packed. It will give better results the sample occupies a greater column depth and getting good resolution of macromolecules within a short time.A symmetrical peak of elution gives the homogeneity of packing.