Biotage® Sfär Flash Columns User Guide

By connecting the tubing, the columns are easily attached to all Biotage®  flash systems. Ensure you remove the Luer-slip adapter to make a secure fitting with the outlet tubing. When needed, keep the Luer-slip adapter on to fit other systems.

Equilibration

Equilibration of flash columns prior to sample addition
» Removes entrapped air
» Eliminates the heat band effect from solvent adsorption on a dry column from the separation run
» Ensures equilibrium between the solvent and the silica.
The minimum recommended equilibration volume is two CV for Biotage®  Sfär columns.

Reversed-Phase Equilibration

Reversed-phase chromatography columns require several steps to ensure they become wet with solvent. Wetting reversed-phase media improves compound retention and therefore loading capacity. For optimal purification with Biotage®  Sfär C18 D and Biotage®  Sfär Bio family columns, follow these steps.
1. Flush with 100% for 3-5 column volumes (CV) of the strong (organic) solvent, typically methanol, acetonitrile or THF.
2. Flush another 3 to 5 CV with 50% strong + 50% aqueous (if system compatible additives are required, include them also at this stage).
3. Flush with another 3 to 5 CV with the initial gradient conditions of your run.