Liquid chromatographic monitoring of pseudocholinesterase activity: comparison of methods.

In order to study the activity of pseudocholinesterase in vitro, two liquid chromatographic techniques have been developed. One is based on phase transfer-catalyzed (PTC) esterification of the carboxylic acid formed during hydrolysis of the substrate, and the other on the use of a radioisotopically labeled substrate. In both cases, the substrate used was a long-chain choline ester. The PTC method, utilizing (N-9-acridinyl)bromoacetamide as a fluorogenic labeling reagent in an aqueous-organic two-phase system, gives esters with very high fluorescence intensity. The radiochromatographic method makes use of on-line radioactivity monitoring of the substrate and product in order to follow the hydrolysis reaction. In both methods reversed-phase liquid chromatography is used. A method for the synthesis of 3H-labeled choline esters is also described. Both techniques are compared with regard to sensitivity, reproducibility and practical considerations.