A novel tris(2,2'-bipyridyl)ruthenium(III) [Ru(biPY)(3)(3+)]-based chemiluminescence (CL) detection in CE using an on-line chemical generation scheme has been demonstrated. Two continuous streams respectively containing solutions of Ru(biPY)(3)(2+) and acidic cerium(IV) used as a homogeneous chemical oxidant are employed to generate Ru(biPY)3, which is delivered into the reaction capillary of a coaxial flow interface and then reacted with analytes at the end of the separation capillary to yield light. The important operational parameters for separation and detection are identified and optimized. Four (alpha-ketocarboxylic acids used as models, outside of the amine-containing compounds, are successfully separated and detected to evaluate the feasibility of the approach. The excellent resolution and detection sensitivity was achieved by using 50 mmol./L phosphate running buffer (pH 9.5) with 0.7 mmol/L CTAB, and CL reagent solution streams containing 0.15 mmol/L Ru(biPY)(3)(2+) and 0.8 mmol/L cerium(IV) (0.25 mol/L H2SO4), respectively. The concentration detection limits for alpha-ketocarboxylic acids were below 3.7 x 10(-8) mol/L (S/N = 3). The proposed method was applied to the determination of (alpha-ketocarboxylic acids in five different honey samples with satisfactory results.