An efficient and highly sensitive chemiluminescence (CL) technique has been developed to detect human serum albumin (HSA). We have previously reported that chemiluminescence produced during interaction between FCLA-1O2 could be modified by the addition of HSA. By carefully selecting the condition of the reaction, the CL intensity could show a linear relationship to the HSA concentration over a clinically interested range of 5times10-9 ~ 8times10-8 mol L-1 ,with a detection limit of 17.32 ng/ml. The method is low cost and has a higher sensitivity than that of fluorescence spectra analysis, synchrony fluorescence scanning technique, and Rayleigh light scattering technique. The technique was used to test HSA in human urine obtained clinically, and the results were consistent to that from other conventional methods. The mechanism of the technique is discussed.