A new chemiluminescence biochemical oxygen demand (BOD(CL)) determining method was studied by employing redox reaction between quinone and Baker's yeast. The measurement was carried out by utilizing luminol chemiluminescence (CL) reaction catalyzed by ferricyanide with oxidized quinone of menadione, and Saccharomyces cerevisiae using a batch-type luminometer. In this study, dimethyl sulfoxide was used as a solvent for menadione. After optimization of the measuring conditions, the CL response to hydrogen peroxide in the incubation mixture had a linear response between 0.1 and 100 microM H2O2 (r2=0.9999, 8 points, n=3, average of relative standard deviation; R.S.D.(av)=4.22%). Next, a practical relationship between the BOD(CL) response and the glucose glutamic acid concentration was obtained over a range of 11-220 mg O2 L(-1) (6 points, n=3, R.S.D.(av) 3.71%) with a detection limit of 5.5 mg O2 L(-1) when using a reaction mixture and incubating for only 5 min. Subsequently, the characterization of this method was studied. First, the BOD(CL) responses to 16 pure organic substances were examined. Second, the influences of chloride ions, artificial seawater, and heavy metal ions on the BOD(CL) response were investigated. Real sample measurements using river water were performed. Finally, BOD(CL) responses were obtained for at least 8 days when the S. cerevisiae suspension was stored at 4 degrees C (response reduction, 69.9%; R.S.D. for 5 testing days, 18.7%). BOD(CL) responses after 8 days and 24 days were decreased to 69.9% and 35.8%, respectively, from their original values (R.S.D. for 8 days involving 5 testing days, 18.7%).