Can I use a reverse-phase HPLC column after I accidentally run the column in the reverse direction? - WKB242325

ENVIRONMENT

• Column characteristics
• Reverse phase
• Reverse direction

ANSWER

It depends on how standards look after returning the column to its normal flow direction.

ADDITIONAL INFORMATION

In some cases, there may be problems with channeling or voiding on some types of  reverse-phase HPLC columns (5 µm, 3.5 µm particles) if they are run in the reverse direction inadvertently. 

To determine if a column will work in the normal flow direction, reinstall the column and make standard injections to see if the peak shape for the standards look Gaussian and if there is good resolution. If peak shape is split or shouldering, this might indicate a potential issue with the packed bed (although there can also be other reasons for peak shape issues that are not related to the packed bed being disrupted).

See: What are common causes of peak splitting when running an LC column?