How do I decrease retention of a very hydrophobic compound on a reverse-phase column? - WKB250345

ENVIRONMENT

• Reverse-phase chromatography
• Hydrophobic compound
• Method development
• Experimental steps

ANSWER

Explore the following steps to elute a very hydrophobic compound from a reverse-phase column:

1) Check if the compound will elute in 100% organic.

2) Test the elution of the compound by increasing the amount of co-solvent in the organic phase (example - adding IPA to acetonitrile to change the elutropic strength of the overall solvent composition).

3) Increase the column temperature.

4) Increase the column flow rate if applicable.

5) Try a less retentive reverse-phase column chemistry (example - a C8, C4, or C1).

6) Try a very different reverse-phase column chemistry with a very different selectivity potential (example - a CSH Fluoro-Phenyl).

7) Try a shorter column length.

ADDITIONAL INFORMATION

For additional information, see Method Development Considerations and Alternative Method Development Techniques

and Exploring Selectivity Benefits of Fluorinated-Phenyl Stationary Phases.