Detection of halogenated hydrocarbon pollutants using enzymatic reflectance biosensor

A reflectometric biosensor for enzymatic determination of halogenated organic pollutant in environmental water sample was developed. Polyacrylate microspheres modified with succinimide functional groups were employed as the substrate matrix for covalent immobilization of haloalkane dehalogenase (DhlA) enzyme, and ETH5294 Nile Blue chromoionophore (NBC) pH dye was physically immobilized on the acrylic micromatrix to function as an optical proton sensor. The hydrolytic dehalogenation of halocarbon catalyzed by the immobilized DhlA resulting in the release of a halogen and a proton, which protonated the immobilized NBC proton indicator, and a colour change of the biosensor from violet to blue was measured with a reflectance spectrophotometer for indirect quantification of the halogenated hydrocarbon concentration. The enzymatic halocarbon reflectance biosensor exhibited a dynamic linear response range of 1–30 mg L−1 1,2-dichloroethane (DCA) (R2 = 0.9762) with a detection limit of 0.3 mg L−1. The proposed enzymatic biosensor gave rapid response, within 2 min, towards the detection of 50 mg L−1 DCA at 662 nm, and demonstrated stable response up to 6 days of storage period at 4 °C and pH 8.0. Validation between the developed biosensor with standard gas chromatography–electron capture detector (GC-ECD) showed that both methods have a comparable accuracy in detecting halocarbon species.