An automated method for the determination of acetyl and pseudo cholinesterase in hemolyzed whole blood

Abstract

The aim of the present study was to develop a method which allows determination of pseudo (PsChE) and acetyl cholinesterase (AChE) activities in single hemolyzed blood samples of workers exposed to cholinesterase-inhibiting compounds, avoiding the time-consuming and laborious separation of plasma and erythrocytes. Two methods based on Ellman's colorimetric determination of cholinesterase activity were compared, and three different substrates were tested. The best results were obtained with the substrates butyrylthiocholine and acetyl(beta-methyl)thiocholine, both showing a substrate specificity of more than 97% with respect to PsChE and AChE, respectively. The method showed sensitivity to detect low levels of inhibition of AChE and PsChE in blood. The between-day precision was less than 4% for both cholinesterase activities. It was demonstrated with this method that hemolyzed blood can be stored at -20 degrees C at least 18 months without loss of cholinesterase activity. The method has been used for 18 months in a monitoring program for laboratory employees working with various cholinesterase-inhibiting compounds. The average co-efficients of intraindividual variation amounted to 6.8% (range 2.2-9.6%; 90 percentile, 8%) and 6.6% (range 2.9-9.9%; 90 percentile, 7.9%) for PsChE and AChE, respectively. In a group of non-exposed workers the average intraindividual variations were 4.0% (range 1.5-7.7%; 90 percentile, 7.6%) and 3.6% (range 0.6-6.6%; 90 percentile, 5.3%), respectively. Using the value of 4.0%, it appears possible to detect an individual decrease in cholinesterase activity of more than 8% below a baseline based on three determinations. The method can thus be used to detect relatively low levels of cholinesterase inhibition.