Authors: MIROSLAV POHANKA, JANA ZDAROVA KARASOVA, KAMIL KUCA, JIRI PIKULA
Abstract: Multichannel spectrophotometry was performed to assay for paraoxon in spiked beverages. A 96-well microplate was used for this purpose. The measuring protocol was based on inhibition of the enzyme acetylcholinesterase by paraoxon that resulted in decreased or no reaction of the enzyme product thiocholine with Ellman's reagent (5,5'-dithiobis [2-nitrobenzoic acid]). The above assay was practically tested using spiked drinking water, mineral water, and coffee. Analytical parameters such as the limit of detection, time, and sample size consumption were adequate. The limit of detection for beverages ranged from 32 to 48 ppb corresponding to 0.32-0.48 ng of paraoxon in absolute values. The described assay seems to be convenient in terms of practical use.
Keywords: Acetylcholinesterase, cholinesterase, biosensing, pesticides, assay, Ellman.
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