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It has been shown that benzethonium chloride produces linear mixed-type inhibition of choline esterase and acetylcholine esterase. These enzymes also show-reagent-carry-over inhibition if the enzyme activities are measured in plastic cuvettes in which previously protein has been determined by the alkaline benzethonium chloride method. Choline esterase is about 10-fold more sensitive to benzethonium chloride than acetylcholine esterase. With acetylthiocholine as substrate Michaelis-Menten constants for choline esterase and acetylcholine esterase are 85 mumol/l and 102 mumol/l, respectively. Carry-over inhibitory effect of benzethonium chloride can be avoided by washing the cuvettes, after protein determination by the benzethonium chloride method, with 5 ml/l Triton X-100, 5 ml/l Tween 20 or 10 g/l sodium dodecyl sulphate. The latter has a disadvantage in that it precipitates out at low temperatures. The dry slide method (Johnson &amp; Johnson) for serum choline esterase is free of the inhibitory effect until the concentration of benzethonium chloride in the sample reaches about 200 mumol/l.

clinical chemistry and laboratory medicine

Inhibition of Acetylcholine Esterase and Choline Esterase by Benzethonium Chloride and Avoidance of the Benzethonium Chloride Carry-Over Inhibitory Effect