title: Quantitative analysis of organophosphorus pesticides in freshwater using an optimized firefly luciferase-based coupled bioluminescent assay.
abstract: In this paper, a coupled bioluminescent assay, relying on the coupling of the enzymes acetylcholinesterase, S-acetyl-coenzyme A synthetase and firefly luciferase, for the detection and quantitation of organophosphorus pesticides, is presented. Using malathion as a model organophosphorus pesticide, the assay was optimized through statistical experimental design methodology, namely Plackett-Burman and central composite designs. The optimized method requires only 20 μL of sample. The linear range for the assay was 2.5-15 μM of malathion, with limits of detection and quantitation of 1.5 and 5.0 μM, respectively. This simple, fast and robust method allows samples to be analyzed at room temperature and without any pretreatment.
authors.affiliation: Centro de Investigação em Química da Universidade do Porto (CIQ-UP), Department of Chemistry and Biochemistry, University of Porto, Porto, Portugal.
authors.forename: S M
authors.initials: SM
authors.lastname: Marques
authors.forename: J C G
authors.initials: JC
authors.lastname: Esteves da Silva
mesh_terms: D005618:Fresh Water / Q000032:analysis; D049409:Luciferases, Firefly / Q000737:chemistry*; D008163:Luminescent Measurements / Q000379:methods*; D009943:Organophosphorus Compounds / Q000032:analysis*; D010575:Pesticides / Q000032:analysis*; D014874:Water Pollutants, Chemical / Q000032:analysis*
chemical_list: D009943:Organophosphorus Compounds; D010575:Pesticides; D014874:Water Pollutants, Chemical; D049409:Luciferases, Firefly
keywords: bioanalytical chemistry; bioluminescence; enzymatic assay; experimental design; luminometry