Why use the empty column volume calculation for equilibration and cleaning of a column and not the actual volume of solvent in a packed column? - WKB117069

ENVIRONMENT

• Column equilibration and cleaning
• Method development
• Method transfer

ANSWER

We suggest calculating the empty column volume to accomplish the following:

• Maximize exposure to as much mobile phase volume that is needed to adequately equilibrate a column and/or clean it.
• Minimize column-to-column and system-to-system method transfer variations. Columns, depending on what they are exposed to, can and will equilibrate faster or slower, even between gel batches. In addition, columns are put on different systems, so technicians do not always take into account delays produced by system module components or tubing/timing, which, when not accounted for, also contribute to robustness issues for chromatographic methods.

ADDITIONAL INFORMATION

Columns will be exposed to whatever is flushing into them—from mobile phase to buildup from used or fouled systems. It is never a bad thing to have more equilibration time or cleaning time to minimize chromatographic differences.

For additional information, see How do I determine column void volume?