Acrylpimaric acid ethylene glycol acrylate was used as the cross-linking agent to prepare rosin-based polymer microspheres through membrane emulsification–microsuspension polymerization. The prepared microspheres were used as the stationary phase in high-performance liquid chromatography for the separation of polycyclic aromatic hydrocarbons (PAHs) and two analogues of alkaloids. Five PAHs (benzene, naphthalene, fluorene, pyrene, and benzo(e)pyrene) were well separated in the rosin-based

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... column with the resolution (Rs) of 6.088 ± 0.006, 5.759 ± 0.017, 7.019 ± 0.020, and 8.584 ± 0.063. A linear relationship was observed for the carbon numbers of PAHs and lnK as obtained with the equation of ln k = 0.22nc – 2.45, R2 = 0.996 . The rosin-based polymer columns were also employed to separate theophylline–caffeine and camptothecin–10-hydroxy-camptothecin, and the good results were obtained with the Rs of 4.617 ± 0.005 and 2.245 ± 0.049, respectively.