Separation of phenylpropionic acids both by strong anion exchange stationary phase and strong cation exchange...

This study demonstrates the expanded application of ion-exchange stationary phases (including strong cation exchange, SCX and strong anion exchange, SAX) in supercritical fluid chromatography (SFC), and more importantly, provides a deeper understanding of the retention mechanisms of these two stationary phases when using the same acidic additive. Phenylpropionic acid compounds (belonging to phenolic acids) were selected as probes. On the SCX column, the π-π and polar interactions originating from the bonded benzenesulfonic acid groups were important foundations for prolonging the retention time of solutes, but they were also the main reason for solutes' tailing profiles. It was found that adding 0.1 % phosphoric acid can generate sufficient strength of electrostatic repulsion to obtain satisfactory peak shapes. Here, phosphoric acid can be adsorbed on the surface of the stationary phase to accumulate more negative charges, and at the same time, the phosphate anion and the phenylpropionic acid in mobile phase might combine into the contact ion pair (CIP)- like through the polar interactions to form the apparently negatively charged solute. On the SAX column, phosphate anions generated by ionization of phosphoric acid, were considered as counterions to effectively mask the electrostatic attraction of quaternary ammonium groups, to significantly reduce the retention of phenylpropionic acids, but improve their resolution. Finally, utilizing two developed SFC methods, i.e., SCX with MeOH + 0.1 % phosphoric acid in CO₂, or SAX with MeOH + 0.3 % phosphoric acid in CO₂, the baseline separation of the extract of Lonicerae Japonicae Flos was achieved within 6 min and 10 min, respectively.