Abstract
A continuous flow method for the rapid measurement of cholinesterase in blood serum was examined with the use of choline-sensitive electrode (C.S.E.) as the amperometric detector. A C.S.E. was constructed by crosslinking both choline oxidase and catalase with bovine serum albumin using glutaraldehyde on a platinum sheet silanized with 3-aminopropyltriethoxysilane. The continuous flow system comprised a peristaltic pump, a line-sample injector, a reaction coil to allow hydrolysis of acetylcholine in phosphate buffer at pH 7.0 in the carrier stream, and the C.S.E. in a flow cell. The C.S.E. was based on the amperometric detection of dissolved oxygen consumed by the overall enzyme reactions.Acetylcholine cholinesterase→ Choline + Acetic acidCholine +O2 choline oxidase-catalase→ Betaine + H2OThe peak current was linearly related to the cholinesterase activity in the range of 1.0 × 1030.8 × 10-1 U; 4050 samples per h could be processed with a relative standard deviation below 3 % without interferences. The electrode retained most of its original activity after repetitive use for two months.
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