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In vivo effects of reversible inhibitors of cholinesterase activity were determined radiometrically in undiluted samples of erythrocytes and plasma. [14C]acetylcholine at substrate saturation, 25 degrees C and pH 7.4 permitted rapid and precise determination of butyrylcholinesterase (EC 3.1.1.8) and acetylcholinesterase (EC 3.1.1.7) activities. Reference values for acetylcholinesterase and butyrylcholinesterase were estimated in the plasma and erythrocyte haemolysate of 102 healthy volunteers. The time course of in vitro inhibition was monitored, starting immediately after addition of 9-amino-1,2,3,4-tetrahydroacridine (tacrine), eserine or pyridostigmine to undiluted human plasma. Maximal inhibition (in vitro) was seen within 60 min with tacrine and eserine, in contrast to 180 min with pyridostigmine. The inhibition remained constant for more than 10 h except with eserine, from which enzyme activity showed an early recovery. Concentration response experiments were performed in undiluted human plasma and undiluted human erythrocyte haemolysate. Ki-values of tacrine, eserine and pyridostigmine were estimated. In contrast to pyridostigmine and eserine, tacrine was found to have a higher affinity for butyrylcholinesterase than for acetylcholinesterase. Tacrine at 2.5 mumol/l resulted in complete inhibition of butyrylcholinesterase and 70% inhibition of acetylcholinesterase activity. Dilution of these samples up to 100-fold was accompanied by almost complete recovery of acetylcholinesterase and by 50% recovery of butyrylcholinesterase.

clinical chemistry and laboratory medicine

Estimation of Cholinesterase Activity (EC 3.1.1.7; 3.1.1.8) in Undiluted Plasma and Erythrocytes as a Tool for Measuring In Vivo Effects of Reversible Inhibitors