Detection of halogenated hydrocarbon pollutants using enzymatic reflectance biosensor

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Abstract

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.

Original languageEnglish
Pages (from-to)80-89
Number of pages10
JournalSensors and Actuators, B: Chemical
Volume281
DOIs
Publication statusPublished - 15 Feb 2019

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Halogenated Hydrocarbons
bioinstrumentation
Biosensors
contaminants
Halocarbons
halocarbons
hydrocarbons
reflectance
Protons
haloalkane dehalogenase
protons
succinimides
Dehalogenation
Ethylene Dichlorides
acrylic resins
Halogens
Organic pollutants
Spectrophotometers
spectrophotometers
Polyacrylates

Keywords

  • Enzymatic biosensor
  • Haloalkane dehalogenase
  • Halogenated hydrocarbon
  • Optical biosensor
  • Reflectance spectrophotometry

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Instrumentation
  • Condensed Matter Physics
  • Surfaces, Coatings and Films
  • Metals and Alloys
  • Electrical and Electronic Engineering
  • Materials Chemistry

Cite this

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title = "Detection of halogenated hydrocarbon pollutants using enzymatic reflectance biosensor",
abstract = "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.",
keywords = "Enzymatic biosensor, Haloalkane dehalogenase, Halogenated hydrocarbon, Optical biosensor, Reflectance spectrophotometry",
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AU - Shahar, Hidayah

AU - Tan @ Chong, Ling Ling

AU - Goh, Choo Ta

AU - Lee, Yook Heng

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N2 - 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.

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