A rapid, sensitive, and practical CE with C4D detection was developed for the analysis of three polyphenols (rutin, scopoletin, and chlorogenic acid) in tobacco samples. The constructed mini detection cell (12 mm x 10 mm x 10 mm) of C4D featured with small inner cell volume (similar to 2 nL), smaller noise (<0.9 mV), repeatability, high strength and durableness. Three polyphenols were ultrasonically extracted with methanolwater (70:30, v/v) solution following SPE cleanup. The CE method was optimized with the running buffer of 150 mmol L-1 2-amino-2-methyl-1-propanol (pH 11.2), and the applied separation voltage of +20 kV over a capillary of 50 mu m id x 375 mu m od x 50 cm (38 cm to the C4D window, 41.5 cm to the UV detector window), which gave a baseline separation of three polyphenols within ca. 6 min. The method provided the limits of quantification (S/N = 10) at about 0.080.15 mu g g-1 for three polyphenols, whereas the overall recoveries ranged from 82% to 88%. The proposed method has been successfully applied to measure three polyphenols in the actual tobacco samples, and their contents were calculated and evaluated.