Optical enzymatic biosensor membrane for rapid in situ detection of organohalide in water samples

An optical biosensor employing immobilized haloalkane dehalogenase (HLD), the halide degrading enzyme for the detection of halogenated organic in environmental water and drinking water samples was developed. The enzymatic biosensor was fabricated by incorporating H+ ion selective chromoionophore ETH5294 and HID enzyme in a stacked chitosan films system on a glass slide. Hydrolytic dehalogenation of dichloroethane (DCA) by the carbon-halide degrading HLD enzyme resulting in the release of a halogen, a proton and a primary alcohol. The halocarbon concentration was optically transduced by the pH transducer layer as a result of protonation reaction of the chromoionophore pH indicator dye embedded in the underneath layer. The resulting colour change of the protonated chromoionophore was measured by fiber optic reflectance spectrophotometry method. Under optimized conditions the detection limit of the proposed reflectance-based enzymatic biosensor membrane was estimated to be 1 mg L-1 with a wide dynamic linear concentration range of 5-60 mg L-1 DCA (R-2 = 0.9792) and satisfactory reproducibility within the relative standard deviation (RSD) range of 3.4-4.3%. Validation test demonstrated that the optical halocarbon biosensor could be a promising tool for rapid (6 min) in situ and direct evaluation of organohalide in river water, tap water and bottled water samples without any sample pre-treatment or extraction steps.