Determination of acetylcholinesterase and butyrylcholinesterase activity without dilution of biological samples
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Miroslav Pohanka
Abstract
Two cholinesterases: acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), are known. The enzymes are important in the body and alteration of their activity has significant use in the diagnosis of poisoning, liver function, etc. Currently available methods for the determination of cholinesterases have some major drawbacks including various interferences and the inability to be used for decreasing the enzyme activity in the presence of reversible inhibitors due to sample dilution; hence, a method for dilution free assay of cholinesterases is desired. Here, microplates were modified with indoxylacetate (100 μL of 10 mmol L−1 solution) and used for cholinesterases assay after drying at 37◦C. The fact that indoxylacetate remains stable in dry state and serves simultaneously as a chromogen and substrate provide good prerequisites for the method. The limit of detection for BChE was 0.71 U while that for AChE was 2.8 U per a 100 μL sample (solution of enzyme or plasma sample). The limit of detection is low enough to allow standard examination of cholinesterasemia. The two cholinesterases can be distinguished from each other using selective inhibitors such as donepezil and iso-OMPA. The new method was also successfully validated for the standard Ellman’s assay using plasma samples with BChE activity adjusted by carbofuran. The new method based on indoxylacetate seems promising for routine tests.
References
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© Institute of Chemistry, Slovak Academy of Sciences
Articles in the same Issue
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- Synthesis, biological evaluation, quantitative-SAR and docking studies of novel chalcone derivatives as antibacterial and antioxidant agents
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Articles in the same Issue
- Deferoxamine–paper for iron(III) and vanadium(V) sensing
- Integrated investigations for the characterisation of Roman lead-glazed pottery from Pompeii and Herculaneum (Italy)
- Determination of acetylcholinesterase and butyrylcholinesterase activity without dilution of biological samples
- Characterization of a novel Aspergillus niger beta-glucosidase tolerant to saccharification of lignocellulosic biomass products and fermentation inhibitors
- Immobilisation of tyrosinase on siliceous cellular foams affording highly effective and stable biocatalysts
- Displacement washing of soda rapeseed pulp
- Hydrovisbreaking of vacuum residue from Russian Export Blend: influence of brown coal, light cycle oil, or naphtha addition
- Antimicrobial properties and chemical composition of liquid and gaseous phases of essential oils
- Syntheses, structures and properties of isonicotinamidium, thionicotinamidium, 2- and 3-(hydroxymethyl)pyridinium nitrates
- Density of lithium fluoride–lithium carbonate-based molten salts
- Synthesis and antimicrobial activity of sulphamethoxazole-based ureas and imidazolidine-2,4,5-triones
- Synthesis, biological evaluation, quantitative-SAR and docking studies of novel chalcone derivatives as antibacterial and antioxidant agents
- Application of polypyrrole nanowires for the development of a tyrosinase biosensor
- Synthesis of a sialic acid derivative of ristocetin aglycone as an inhibitor of influenza virus
- Erratum to “Ľubomír Vančo, Magdaléna Kadlečíková, Juraj Breza, Pavol Michniak, Michal Čeppan, Milena Reháková, Eva Belányiová, Beata Butvinová: Differentiation of selected blue writing inks by surface-enhanced Raman spectroscopy”, Chemical Papers 69 (4) 518–526 (2015)
