Determination of acetylcholinesterase and butyrylcholinesterase activity without dilution of biological samples

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60162694%3AG44__%2F15%3A43875381" target="_blank" >RIV/60162694:G44__/15:43875381 - isvavai.cz</a>

Výsledek na webu

<a href="http://www.degruyter.com/view/j/chempap.2015.69.issue-8/chempap-2015-0117/chempap-2015-0117.xml" target="_blank" >http://www.degruyter.com/view/j/chempap.2015.69.issue-8/chempap-2015-0117/chempap-2015-0117.xml</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1515/chempap-2015-0117" target="_blank" >10.1515/chempap-2015-0117</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Determination of acetylcholinesterase and butyrylcholinesterase activity without dilution of biological samples

Popis výsledku v původním jazyce

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 mu L of 10 mmol L-1 solution) and used for cholinesterases assay after drying at 37 degrees 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 mu L 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. (C) 2015 Institute of Chemistry, Slovak Academy of Sciences

Název v anglickém jazyce

Determination of acetylcholinesterase and butyrylcholinesterase activity without dilution of biological samples

Popis výsledku anglicky

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 mu L of 10 mmol L-1 solution) and used for cholinesterases assay after drying at 37 degrees 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 mu L 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. (C) 2015 Institute of Chemistry, Slovak Academy of Sciences

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CB - Analytická chemie, separace

OECD FORD obor

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Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2015

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Chemical Papers-Chemicke Zvesti

ISSN

0366-6352

e-ISSN

—

Svazek periodika

69

Číslo periodika v rámci svazku

8

Stát vydavatele periodika

SK - Slovenská republika

Počet stran výsledku

6

Strana od-do

1044-1049

Kód UT WoS článku

000354752200004

EID výsledku v databázi Scopus

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