VZL 2010, 79(3):111-113

Testing of a Referential Method Based on Indoxylacetate as a Acetylcholinesterase Substrate for Identification of Reactivation Efficacy

Miroslav Pohanka ORCID...1,2, Martina Hrabinová ORCID...2, Jana Žďárová Karasová ORCID...2, Ondřej Holas3, Kamil Kuča ORCID...1,2, Jiří Cabal2
1 Univerzita obrany, Fakulta vojenského zdravotnictví, Centrum pokročilých studií, Hradec Králové
2 Univerzita obrany, Fakulta vojenského zdravotnictví, katedra toxikologie, Hradec Králové
3 Univerzita Karlova v Praze, Farmaceutická fakulta v Hradci Králové, katedra farmaceutické chemie, Hradec Králové

Biochemické vyhodnocení aktivity acetylcholinesterasy je důležitým faktorem při výběru a hodnocení látek modulující její aktivitu, jako jsou oximové reaktivátory. Úskalím v současnosti používané Ellmanovy metody pro stanovení aktivity cholinesteras je samovolná reakce mezi Ellmanovým činidlem a oximovým reaktivátorem. V této práci je použit a popsán alternativní chromogenní substrát indoxylacetát umožňující vyhodnotit účinnost reaktivace acetylcholinesterasy bez znatelné interference in vitro. Metoda byla úspěšně ověřena pro pět standardních oximových reaktivátorů (HI-6, methoxim, trimedoxim, obidoxim, 2-PAM) a tři organofosfátové inhibitory: paraoxon-methyl, sarin a tabun.

Keywords: Organofosfátový insekticid; Nervově paralytická látka; Acetylcholinesterasa; Reaktivátor; Indoxylacetát

Biochemical evaluation of acetylcholinesterase activity is an important factor for selection and evaluation of drugs, such as oxime reactivators, which modulate acetylcholinesterase activity. A spontaneous reaction between an Ellman reagent and an oxime reactivator is the main disadvantage of currently available Ellman's method used for identification of cholinesterase activity. In this study, an alternative chromogen, substrate indoxylacetate is used. It is suitable for evaluation of acetylcholinesterase reactivation efficacy without oxime interference in vitro. The method was successfully examined for five standard oxime reactivators (HI-6, methoxime, trimedoxime, obidoxime, and 2-PAM) and three organophosphate inhibitors: paraoxon-methyl, sarin and tabun.

Keywords: Organophosphate insecticide; Nerve agent; Acetylcholinesterase; Reactivator; Indoxylacetate

Received: December 15, 2009; Published: September 1, 2010  Show citation

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Pohanka, M., Hrabinová, M., Žďárová Karasová, J., Holas, O., Kuča, K., & Cabal, J. (2010). Testing of a Referential Method Based on Indoxylacetate as a Acetylcholinesterase Substrate for Identification of Reactivation Efficacy. Vojenské Zdravotnické Listy, 79(3), 111-113
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