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What steps should be taken to extend the life of a reversed-phase HPLC column?
The answer to the following question was provided by LCGC’s “LC Troubleshooting” columnist John Dolan.
Q: What steps should be taken to extend the life of a reversed-phase HPLC column?
A:John Dolan: Here’s a short list of the basics:
• Perform sample cleanup and filtration or centrifugation
• Place an inline filter between the autosampler and column with smaller frit porosity than that of the column frit (for example, 0.5 µm is good for 3- and 5-µm particles columns)
• Use a guard column (optional)
Keep it clean:
• Flush with the strong solvent of the mobile phase (for example, acetonitrile or methanol) after each batch of samples
• Backflush if necessary to remove particulates from frit (shouldn't happen if you take the steps in “Protect it”)
Use dedicated columns:
• One column per method, even if a different method calls for the same column
Remember that the column is a consumable item:
• After you reach ~500 injections, the cost per sample from the column is