Mosca A, Onelli E, Rosti E, Paleari R, Luzzana M, Imbimbo B P
Department of Sciences and Biomedical Technologies, University of Milan, Italy.
Ther Drug Monit. 1995 Jun;17(3):230-8. doi: 10.1097/00007691-199506000-00004.
Rapid and reliable measurement of acetylcholinesterase (AChE) activity is of crucial importance to the pharmacodynamic monitoring of anticholinesterase drugs. A new assay has been developed to measure AChE from 10 microliter samples of capillary blood. AChE activity was calculated from the change in pH of the reaction medium caused by the hydrolysis of acetylcholine and measured with a highly sensitive differential pH apparatus (CL-10, Eurochem, Rome, Italy). Interference by butyrylcholinesterase was eliminated by a specific inhibitor, quinidine sulfate. The assay lasts 1 min. The coefficient of variation (CV) for replicated measurements was 2.8% (3267 U/L, n = 33). Linearity ranged from 0 to 10,000 U/L. The correlation coefficient between the new technique and Ellman's colorimetric method on washed erythrocytes was r = 0.987 (y = 1.299x - 63, n = 29). The correlation coefficient between assays on capillary and venous samples was r = 0.979 (y = 0.974x + 174, n = 47). A cross-laboratory validation study was performed in 10 centers using glycerol-stabilized hemolysates with normal and reduced AChE activity. Samples were assayed in triplicate. The within- and between-laboratory CVs for samples with normal AChE activity (6,018 U/L) were 2.2 and 8.1%, respectively. The new method was applied to a double-blind, placebo-controlled multicenter study of eptastigmine in Alzheimer patients and proved to be a simple, noninvasive, rapid, and reliable method for pharmacodynamic monitoring of this drug.
快速、可靠地测定乙酰胆碱酯酶（AChE）活性对于抗胆碱酯酶药物的药效学监测至关重要。现已开发出一种新的检测方法，用于测定10微升毛细血管血样中的AChE。通过乙酰胆碱水解引起的反应介质pH变化来计算AChE活性，并使用高灵敏度差分pH仪（CL - 10，Eurochem，意大利罗马）进行测量。通过特定抑制剂硫酸奎尼丁消除丁酰胆碱酯酶的干扰。该检测耗时1分钟。重复测量的变异系数（CV）为2.8%（3267 U/L，n = 33）。线性范围为0至10,000 U/L。新技术与洗过红细胞的埃尔曼比色法之间的相关系数为r = 0.987（y = 1.299x - 63，n = 29）。毛细血管样本和静脉样本检测之间的相关系数为r = 0.979（y = 0.974x + 174，n = 47）。在10个中心进行了一项跨实验室验证研究，使用具有正常和降低AChE活性的甘油稳定溶血产物。样本进行了三次检测。正常AChE活性（6,018 U/L）样本的实验室内和实验室间CV分别为2.2%和8.1%。该新方法应用于一项关于艾斯他明治疗阿尔茨海默病患者的双盲、安慰剂对照多中心研究，结果证明它是一种用于该药物药效学监测的简单、无创、快速且可靠的方法。
