Optimize the separation of fluorinated amphiles using high-performance liquid chromatography

•The elution order of the analytes corresponds to their eluent-philicity.•Optimal separation was achieved through hetero-pairing of column and eluent.•Separation is further improved by elevated temperature.•Separation in fluorous mixture synthesis is based on the fluorine content percentage F% rather than the number of fluorine atoms.

Using the set of fluorinated amphiles that contain the same fluorocarbon moiety but differ in their fluorine content percentage F% (25–45%), the optimal condition for a F%-based separation of these analytes using reverse-phase chromatography was explored. It is found that optimal separation can be achieved by pairing a regular reverse-phase column (such as C8) with a fluorinated eluent (such as trifluoroethanol). Separation is further improved at higher chromatographic temperature with baseline separation achieved at 45 °C. This result indicates that the separation of fluorocarbon-tagged molecules can be based on the fluorine content percentage rather than the number of fluorine atoms.

Graphical abstractOptimal separation of fluorinated amphiles with HPLC can be achieved by pairing a regular reverse-phase column with a fluorinated eluent and separation is further improved by elevated temperature.Figure optionsDownload full-size imageDownload as PowerPoint slide