Chelminska-Bertilsson M, Allenmark S
Laboratory of Microbiological Chemistry, University of Gothenburg, Sweden.
J Chromatogr. 1992 Mar 27;575(2):237-42. doi: 10.1016/0378-4347(92)80151-f.
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.
为了研究体外假性胆碱酯酶的活性，已开发出两种液相色谱技术。一种基于底物水解过程中形成的羧酸的相转移催化（PTC）酯化反应，另一种基于使用放射性同位素标记的底物。在这两种情况下，所使用的底物都是长链胆碱酯。PTC方法在水-有机两相系统中利用（N-9-吖啶基）溴乙酰胺作为荧光标记试剂，生成具有非常高荧光强度的酯。放射色谱法利用对底物和产物的在线放射性监测来跟踪水解反应。两种方法均使用反相液相色谱。还描述了一种合成3H标记胆碱酯的方法。对这两种技术在灵敏度、重现性和实际考虑因素方面进行了比较。
