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:
Protect it:
• 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
Modern HPLC Strategies: Improving Retention and Peak Shape for Basic Analytes
August 16th 2024In high-performance liquid chromatography (HPLC), it is common for bases and unreacted ionized silanols on silica-based columns to cause irreproducible retention, broad peaks, and peak tailing when working with basic analytes. David S. Bell, Lead Consultant at ASKkPrime LLC offers innovative HPLC strategies that can help mitigate such issues.