Preparation of mixed-mode chromatographic stationary phases based on modified rosin and its application in the separation of natural medicines

Biomass materials, such as rosin obtained from pine trees, are increasingly employed in developing high-performance liquid chromatography (HPLC) stationary phase due to their environmental sustainability, abundant resources, cost-effectiveness and renewability. In this study, we synthesized a novel mixed-mode chromatographic stationary phase (Sil-HRE-DPM) with reversed-phase, hydrophilic, and weak anion-exchange functionalities via thiol-ene click chemistry by copolymerizing hydrogenated rosin acrylic glycol ester (HRE) and N-(3-dimethylaminopropyl)methacrylamide (DPM). The successful synthesis of Sil-HRE-DPM was confirmed through comprehensive characterizations. Sil-HRE-DPM demonstrated excellent chromatographic stability, repeatability and excellent efficiency (71,796 plate/m). The retention mechanism of Sil-HRE-DPM was examined utilizing the linear solvation energy relationship (LSER) model and thermodynamic analysis. Various probes, including Tanaka test mixtures, hydrophobic and hydrophilic analytes, and anionic chemicals were employed to evaluate retention mechanisms and separation performance. It was found that Sil-HRE-DPM provides various interactions, such as hydrophobic, π-π, hydrophilic and weak anionic exchange. Finally, the column was effectively applied to separate the active components of the natural medicines, specifically Erigeron breviscapus, herb Evodia rutaecarpa and Panax notoginseng . This study provides a theoretical foundation for the quality control of traditional Chinese medicine and the efficient utilization of rosin, thereby facilitating advancements in both pharmaceutical and industrial applications.