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Do you have a recommended column cleaning procedure for reversed phase columns?

GENERAL CARE AND USE

HPLC UHPLC

Last Updated: 12/2/2014

 

The following can be used as a guideline for regeneration/column cleaning for many reversed phase HPLC columns. 

Rinse with 10 Column Volumes each of:

  • 95 % Water/5% Acetonitrile (for buffer removal)
  • THF
  • 95% Acetonitrile/5% Water
  • Mobile Phase​

For instances of high back pressure it is permissible to "reverse- flush" Phenomenex columns. This practice should not be used on other manufacturers' columns, which may not be to similar standards.

Before starting any kind of cleaning procedure, make sure your in-column solvent or mobile phase is miscible with the recommended cleaning solvents. Also, flow-rates should be 1/5-/12 of the typical flow-rate

Column volume can be estimated using the following equation:

V=πr²L

  • ​V= column volume in mL
  • r= column radius in cm
  • L= column length in cm