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Selective monitoring of enzymatic activity of acetylcholinesterase by flow injection analysis with mass spectrometric detection

Vlastimil Dohnal, Lucie Dohnalova, Daniel Jun, Kamil Kuca

. 2013 ; 82 (3) : 120-125.

Language English Country Czech Republic

Persistent link   https://www.medvik.cz/link/bmc14043929

Flow injection analysis (FIA) is an analytical method where the reaction mixture is injected into flow of liquid. The reaction product is monitored by a suitable detector such as ultraviolet/visible (UV/VIS) spectrophotometric or electrochemical detector. Mass spectrometric detectors (MS) are coming to be a standard equipment of analytical laboratories in the present time. This work is focused on application of FIA-MS instrumentation for monitoring of Ellman's reaction where both reactants (acetylthiocholine and 5,5’-dithiobis-2-nitrobenzoic acid, DTNB) and the reaction product (5-mercapto-2-nitrobenzoic acid) are monitored. This reaction is usually used for monitoring of acetylcholinesterase and butyrylcholinesterase. Due to its simplicity, the developed method is generally applicable for monitoring of enzymatic reactions of cholinesterases. The main advantage of this method is high selectivity and reduction of influence of compounds, which are reacting with DTNB, resulting in a color product of Ellman's reaction.

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Literatura

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$a Dohnal, Vlastimil $7 pna2015873045 $u 1 Department of Chemistry, Faculty of Science, University of Hradec Kralove, Rokitanskeho 62, 500 03 Hradec Kralove, Czech Republic
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$a Selective monitoring of enzymatic activity of acetylcholinesterase by flow injection analysis with mass spectrometric detection / $c Vlastimil Dohnal, Lucie Dohnalova, Daniel Jun, Kamil Kuca
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$a Flow injection analysis (FIA) is an analytical method where the reaction mixture is injected into flow of liquid. The reaction product is monitored by a suitable detector such as ultraviolet/visible (UV/VIS) spectrophotometric or electrochemical detector. Mass spectrometric detectors (MS) are coming to be a standard equipment of analytical laboratories in the present time. This work is focused on application of FIA-MS instrumentation for monitoring of Ellman's reaction where both reactants (acetylthiocholine and 5,5’-dithiobis-2-nitrobenzoic acid, DTNB) and the reaction product (5-mercapto-2-nitrobenzoic acid) are monitored. This reaction is usually used for monitoring of acetylcholinesterase and butyrylcholinesterase. Due to its simplicity, the developed method is generally applicable for monitoring of enzymatic reactions of cholinesterases. The main advantage of this method is high selectivity and reduction of influence of compounds, which are reacting with DTNB, resulting in a color product of Ellman's reaction.
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$a Dohnalová, Lucie $7 _AN076555 $u Department of Military Hygiene, Faculty of Military Health Sciences, University of Defence, Trebesska 1575, 500 01 Hradec Kralove, Czech Republic
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$a Jun, Daniel, $d 1976- $7 xx0040498 $u Biomedical Research Center, University Hospital Hradec Kralove, Sokolska 581, 500 05 Hradec Kralove, Czech Republic; Center of Advanced Studies, Faculty of Military Health Sciences, University of Defence, Trebesska 1575, 500 01 Hradec Kralove, Czech Republic
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$a Kuča, Kamil, $d 1978- $7 xx0041831 $u Biomedical Research Center, University Hospital Hradec Kralove, Sokolska 581, 500 05 Hradec Kralove, Czech Republic; Center of Advanced Studies, Faculty of Military Health Sciences, University of Defence, Trebesska 1575, 500 01 Hradec Kralove, Czech Republic
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