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Quantitative analysis of organophosphorus pesticides in freshwater using an optimized firefly luciferase‐based coupled bioluminescent assay
Author(s) -
Marques SM,
Silva JCG Esteves
Publication year - 2014
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/bio.2556
Subject(s) - luciferase , bioluminescence , detection limit , firefly protocol , chromatography , chemistry , malathion , acetylcholinesterase , pesticide , linear range , luciferases , enzyme , biochemistry , biology , zoology , transfection , agronomy , gene
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. Copyright © 2013 John Wiley & Sons, Ltd.