Novel post-functionalized polymer microspheres containing capsaicin as reversed-phase chromatography stationary phase separation for hydrophilic compounds

[Display omitted] •The highly stable polymer microspheres were copolymerized, whose size was 2 μm.•Microspheres had the potential to be produced in large quantities.•The spheres were post-functionalized by double coating to form core–shell structure.•The microspheres, as stationary phase, could be efficient for hydrophilic compounds. Here, we created for the first time that highly stable polymer microspheres copolymerized with capsaicin derivative, divinyl benzene, and N-(Hydroxymethyl)acrylamide could be efficient for reversed-phase high-performance liquid chromatography (RP-HPLC) by using a simple “one-pot cooking” method, which had the potential to be produced in large quantities. Then, the spheres were post-functionalized with two different monomers, N-vinyl formamide and 4-acrylomorpholine by double coating, which had ideal mechanical strength and efficiency as stationary phase. In order to reach best separation condition, the effects of buffer concentration, pH, column temperature, and mobile phase polarity on the retention rate were studied. Specifically, the interaction mechanism showed that the stationary phase had hydrophobic and electrostatic interaction with the water-soluble vitamins. It is of significant interest that all columns had excellent precision and accuracy (RSD