Cholinesterase assay automated in the Cobas-Bio centrifugal analyzer

We describe a method for measuring plasma cholinesterase (EC 3.1.1.8) and dibucaine and fluoride numbers by using the Cobas-Bio. Benzoyl choline chloride is used as substrate. Reaction conditions are the same as in the corresponding manual method. The reaction is stopped with physostigmine. Choline oxidase (EC 1.1.3.17) is coupled with 4-aminoantipyrene. In the presence of peroxidase (EC 1.11.1.7), the 2-hydroxy-3,5-dichlorobenzenesulfonate indicator reaction gives a red product, measured at 505 nm. The analyzer is used in the "Multi Run" mode, with plasma cholinesterase and dibucaine and fluoride inhibition concurrently measured for eight samples. Coefficients of correlation between the manual and present method for plasma cholinesterase and dibucaine and fluoride numbers in 40 patients of various phenotypes were 0.843, 0.923, and 0.717, respectively. The inter-batch CV for each of the three assays was 2% to 3%.