Augustinsson K B, Eriksson H, Faijersson Y
Clin Chim Acta. 1978 Oct 16;89(2):239-52. doi: 10.1016/0009-8981(78)90322-4.
A sensitive method, especially suitable for clinical laboratories, for the routine determination of cholinesterase activities in whole blood is presented. This method is based on the hydrolysis of propionylthiocholine and the spectrophotometric determination of the thiocholine produced by reaction with 4,4'-dithiodipyridine. The reaction product 4-thiopyridone has an absorption maximum at 324 nm, so that measurement in the presence of hemoglobin is possible. Propionylthiocholine is used at the substrate for both plasma butyrylcholinesterase and erythrocyte acetylcholinesterase. These two enzymes, in the relative amounts at which they are present in human blood, split this ester at about the same rate. Consequently, a first determination gives the total activity of which each individual activity is about 50%. A second determination in the presence of a selective inhibitor ("Astra 1397") for plasma butyrylcholinesterase gives the activity of the erythrocyte acetylcholinesterase. The difference between the two values represents the activity of the plasma enzyme. The validity of the method and the reliability of the results were checked with each blood sample in two ways: (1) by determining the activities of whole blood with an earlier gasometric technique which uses blood sample dried on filter paper; and (2) by measuring the activities in separated plasma and erythrocyte hemolysate eith propionylthiocholine as the substrate.
本文介绍了一种特别适用于临床实验室的灵敏方法，用于常规测定全血中的胆碱酯酶活性。该方法基于丙酰硫代胆碱的水解以及与4,4'-二硫代二吡啶反应生成的硫代胆碱的分光光度测定。反应产物4-硫代吡啶酮在324nm处有最大吸收，因此可以在血红蛋白存在的情况下进行测量。丙酰硫代胆碱用作血浆丁酰胆碱酯酶和红细胞乙酰胆碱酯酶的底物。这两种酶在人血中的相对含量下，以大致相同的速率分解这种酯。因此，第一次测定得到的总活性中，每种酶的活性约为50%。在存在血浆丁酰胆碱酯酶的选择性抑制剂（“Astra 1397”）的情况下进行第二次测定，可得到红细胞乙酰胆碱酯酶的活性。这两个值之间的差值代表血浆酶的活性。该方法的有效性和结果的可靠性通过两种方式对每个血样进行了检验：（1）使用在滤纸上干燥的血样，通过早期的气体测量技术测定全血的活性；（2）以丙酰硫代胆碱为底物，测量分离的血浆和红细胞溶血产物中的活性。
