A poly(ethyleneimine) embedded phenyl mixed-mode stationary phase was prepared through epoxide ring opening reaction. Elemental analysis (EA), Fourier transform infrared spectroscopy (FT-IR) and thermogravimetric analysis (TGA) were used to characterize whether functional groups have been modified on the surface of silica successfully. The researches of chromatographic performance for designed column were divided to four parts. The retention behavior under reversed-phase mode was studied using polycyclic aromatic hydrocarbons (PAHs), positional isomers and alkylbenzenes. The retention characteristic under hydrophilic interaction mode was studied by nucleosides, nucleobases and flavonoids. The chromatographic performance of anion exchange mode was evaluated by benzoic acids and phenols. Due to a high content of amino and phenyl groups on the stationary phase, aromatic amines were chosen for the evaluation of retention mechanisms of benzene ring attraction and amino repulsion. The investigations about effects of mobile phase constitution and pH on retention demonstrated the mixed-mode retention mechanisms of the column. Furthermore, due to amino selectivity of the column, a method for the rapid separation and determination of phenylenediamines in hair dye was established. And both linear correlation coefficients of p-diaminobenzene and m-diaminobenzene in concentration range from 0.1 to 30 μg mL-1 were over 0.999. In conclude, the prepared mixed-mode stationary phase could realize various separation modes by adjusting chromatographic conditions, and it had the potential for the rapid separation and determination of aniline compounds in various complex samples.
Keywords: Hair dye; Mixed-mode chromatography; Poly(ethyleneimine); Reversed phase/hydrophilic interaction/ion exchange; Stationary phase.
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