ISSN print edition: 0366-6352
ISSN electronic edition: 1336-9075
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A new fluorimetric method for determination of submicrogram amounts of cholinesterase inhibitors

J. Matoušek, J. Fischer, and J. Cerman

The Antonín Zápotocký Military Academy, Brno

 

Abstract: A highly sensitive method for determining organophosphorus compounds on the base of inhibition of choline esterase is described. Indoxyl acetate is used as a substrate, the reaction index being the change in fluorescence at time АФ/At. As preparation of choline esterase a lyophilised horse blood plasm was used. The optimum pH of the enzymatic hydrolysis is at 7.4 — 7.6 (a spontaneous hydrolysis at this pH and 25 °C does not take place). Km of indoxyl acetate at pH 7.6 and 25 °C is 6 . 10-4 м. The rate of the enzymatic hydrolysis increases linearly with concentration of choline esterase. The relative standard deviation for the choline esterase determination s = ± 1.5 %. The sensitivity of determination for O-isopropylmethanenuorophosphonate 1.4 . 10-7mg/ml;O-pinacolylmethanenuorophosphonate 1.10-7 mg/ml;O-ethyl-N,N-dimethylamidocyanophosphate 1.6 . 10-7 mg/ml; O,0'-diisopropylrluorophosphate 1.6 10- 5 mg/ml; O,O'-diethyl-O''-nitrophenylphosphate 2.8 10-7 mg/ml; 0-ethyl-S-2-dimethylaminoethylmethanethiophosphonate 2 . 10-5 mg/ml. Determination time 12—15 minutes.

Full paper in Portable Document Format: 223a184.pdf (in Czech)

 

Chemical Papers 22 (3) 184–189 (1968)

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