ALL CATEGORIES

HPLC Column Cleaning Guide

Are you experiencing unexpected changes in peak shape, retention time, and/or elevated column pressure? This could be a sign that your column is needing cleaned.

The following guidelines describe general indications of column deterioration and column cleaning procedures. For detailed column cleaning procedures, refer to the operating manual packed with each column.

Typical Indicators of Column Deterioration:

  • Elevated column pressure
  • Abnormal peak shape (broadening, fronting, or tailing)
  • Split peaks
  • Change in retention time
  • Unstable baseline

RPC Columns

  • Always use a strong organic solvent such a methanol, acetonitrile or THF for cleaning
  • If a buffered mobile phase has been used, check the miscibility of the buffer solution and the organic solvent before cleaning the column
  • Disconnect any guard column
  • Reverse the direction of the column and disconnect the outlet from the detector
  • Set the flow rate to 0.5mL/min or half of method flow rate
  • Wash the column 3 to 5 time the column volume with 100% organic solvent
  • Wash the column 3 to 5 times the column volume with 100% organic solvent
  • Wash the column with 3 to 5 times the column volume with original mobile phase
  • Repeat the 100% organic step if enhanced cleaning is required

Sugar Analysis Columns (NH2P and VG-50 Series)

In cases where an acidic substance has been bound to the amino functional group, flush with solvents in the following sequence:

  • 100% water
  • 1M perchloric acid (aqueous)
  • 100% water
  • 1M NaOH (aqueous)
  • 100% water
  • New mobile phase

HILIC Chromatography Columns

  • Always use a strong organic solvent such as IPA, methanol or THF for cleaning
  • If a buffered mobile phase has been used, check the miscibility of the buffer solution and the organic solvent before cleaning the column
  • Prepare a 50:50 cleaning solution of solvent and water
  • Disconnect any guard column
  • Reverse the direction of the column and disconnect the outlet from the detector
  • Set the flow rate to 0.5mL/min or half of method flow rate
  • Wash the column 3 to 5 times the column volume with the cleaning solution
  • Wash the column with 3 to 5 times the column volume with original mobile phase
  • Strong buffers may be required to remove ionic contaminants

Download our HPLC Column Guide

For more information and other useful guidance download our HPLC Column Guide.