A new, rapid and sensitive method for the determination of bovine serum albumin (BSA) by flow-injection coupled with chemiLuminescence detection was establish, which based on BSA enhancing effect on weak chemiLuminescence system of Luminol-H2O2 in alkaline solution. Under the optimized conditions, linearity for BSA was found in the range of 2.5×10-9～1.0×10-6 mol·L-1, with the detection limit of 5.8×10-10 mol·L-1. And the sampling detecting rate was about 112 samples·h-1. The relative standard deviation(RSD)was found to be 0.8% for 11 replicate determinations of 1.0×10-7 mol·L-1 BSA. The proposed method has been satisfactorily applied to the determination of BSA in the bovine serum samples. The possible mechanism of Luminol-H2O2-BSA was studied by chemiLuminescence spectrum and UV-visible spectrum.