A spectrophotometric biochemical oxygen demand determination method using 2,6-dichlorophenolindophenol as the redox color indicator and the eukaryote Saccharomyces cerevisiae

Abstract

A method to determine the spectrophotometric biochemical oxygen demand (BOD sp) was studied with high sensitivity and reproducibility by employing 2,6-dichlorophenolindophenol (DCIP) as a redox color indicator, the yeast Saccharomyces cerevisiae, and a temperature-controlling system providing a three-consecutive-stir unit. The absorbance of DCIP decreased due to the metabolism of organic substances in aqueous samples by S. cerevisiae. Under optimum conditions, a calibration curve for glucose glutamic acid concentration between 1.1 and 22 mg O 2 L −1 ( r = 0.988, six points, n = 3) was obtained when the incubation mixture was incubated for 10 min at 30 °C. The reproducibility of the optical responses in the calibration curve was 1.77% (average of relative standard deviations; RSD av). Subsequently, the characterization of this method was studied. The optical responses to pure organic substances and the influence of chloride ions, artificial seawater, and heavy metal ions on the sensor response were investigated before use with real samples. Measurements of real samples using river water were performed and compared with those obtained using the BOD 5 method. Finally, stable responses were obtained for 36 days when the yeast cell suspension was stored at 4 °C (response reduction, 89%; RSD av value for 9 testing days, 8.4%).