Green synthesis of biomass derived monodisperse microspheres used as chromatographic stationary phases for separation of dye molecules in ethanol.

Green liquid chromatography (LC) has garnered increasing attention; however, its adoption as a replacement for conventional methods is limited by inferior separation performance. Herein, we present a fully integrated design for green LC, encompassing biomass-derived starting materials, the green synthesis of a solid stationary phase, and environmentally benign separation using ethanol as the mobile phase. Via a green precipitation polymerization technique (room temperature quiescent redox-initiated precipitation polymerization, RTQRPP), we synthesized a series of monodisperse microspheres derived from natural fatty acids. Notably, microspheres composed of poly(divinylbenzene-co-tetradecyl methacrylate) (P(DVB-TMA))-where tetradecyl methacrylate (TMA) is derived from myristic acid-exhibited a uniform particle size of 4.6 μm (polydispersity index, PDI < 1.01). The resulting P(DVB-TMA) material displays exceptional superoleophilicity and superhydrophobicity, with water contact angles reaching 160 ° Consequently, columns packed with P(DVB-TMA) generated significantly lower back pressure under pure water compared to commercial C18 silica columns. Furthermore, when using an ethanol mobile phase, the P(DVB-TMA) column successfully achieved baseline separation of a four-component dye mixture, demonstrating markedly better resolution than a commercial C18 column under identical conditions. Molecular dynamics simulations and density functional theory calculations confirmed a strong reversed-phase retention mechanism, with elution order correlating with decreasing aqueous solubility (Log S) at 25 ℃. The column reproducibility is proved by that the relative standard deviations (RSD) of retention times can be controlled in 0.40∼0.70 % for the intra-day and 0.77∼1.84 % for the inter-day tests, respectively. Our findings establish this novel stationary phase as a superior, greener alternative to commercial columns for target compound separation.