Abstract: In this study, a chiral polyimine macrocycle, BINOL-CPM, was synthesized via a one-step condensation of (S)-2,2'-dihydroxy-[1,1'-binaphthalene]-3,3'-dialdehyde and 1,2-phenylenediamine. After modification, it was bonded to the surface of thiolated silica via thiol-ene click reaction to fabricate a chiral stationary phase (CSP) for high-performance liquid chromatography (HPLC). The CSP-packed column exhibits good chiral separation performance in both normal-phase (NP) and reversed-phase (RP) elution modes, with 12 racemates resolved in the NP mode and 6 racemates resolved in the RP mode. The chiral separation performance of this column for these resolved racemates was compared with two commonly used commercial columns, Chiralpak AD-H and Chiralcel OD-H. Among the 12 racemates resolved in the NP mode, 4 racemates [1-(3-fluorophenyl)ethanol, 1-phenylethanol, 3-hydroxy-2-butanone, and 1-(4-fluorophenyl)ethanol] could not be resolved on the Chiralpak AD-H column, and 3 racemates [1-(3-fluorophenyl)ethanol, 3-hydroxy-2-butanone, and 1-(4-fluorophenyl)ethanol] could not be resolved on the Chiralcel OD-H column. Among the 6 racemates resolved in the RP mode, 1 racemates (1-phenylethanol) could not be resolved on the Chiralpak AD-H column. Moreover, the resolution values (Rs) of some racemates on this column was higher than that on the two commercial columns. Therefore, this prepared column demonstrates good complementarity in chiral separation with the two commonly used commercial columns, enabling the separation of some chiral compounds that cannot be resolved or not well be resolved by these two commercial columns. In addition, the column also demonstrates excellent reproducibility and stability in chiral separation. After hundreds of injections, the retention time (tR) and Rs of the two enantiomeric peaks for the separation of ethyl mandelate shows no significant changes compared to the initial use of the column. The relative standard deviation (RSD, n = 5) of tR for the chromatographic peaks of the two enantiomers was less than 0.49 %, and the RSD of Rs (n = 5) was less than 2.88 %. Moreover, the column-to-column reproducibility was also good, with the RSD (n = 3) of tR and Rs for the two enantiomeric peaks of ethyl mandelate separation on columns prepared from different batches being less than 1.09 % and 5.75 %, respectively. This study indicates that BINOL-CPM is a promising chiral separation material for HPLC and has good research significance and value. It also provides a reference for the development of novel HPLC CSP.

Key words: High-performance liquid chromatography, Chiral separation, Chiral stationary phase, Chiral polyimide macrocycle