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In this study, the authors performed X-ray photoelectron spectroscopy (XPS) and near-edge X-ray absorption fine structure (NEXAFS) studies of vacuum ultraviolet (VUV)/O3-treated aromatic polyurea films to investigate their treatment effects. XPS and NEXAFS spectra indicate that the benzene ring was cleaved after treatment and that carboxyl, hydroxyl, ketone and aldehyde groups were formed at the cleaved sites. The VUV/O3-treated polyurea film was applied to a polymethylmethacrylate (PMMA) microchip for microchip electrophoresis (MCE) of bovine serum albumin (BSA). Fast electro-osmotic mobility of 4.6 × 10-4 cm2/V/s as well as reduction of the BSA adhesion was achieved. This functional surface is useful for high-speed MCE analysis.