DIUTHAME (desorption ionization using through hole alumina membrane) is a novel matrix-free laser desorption/ionization method that enables highly reproducible acquisition of mass spectra. This study aims to evaluate the applicability of DIUTHAME to the acetylcholinesterase reaction assay (AChE assay) commonly used in high-throughput screening (HTS) in the drug discovery process. The commercially available 9-ch DIUTHAME chip was applied to a series of AChE assays prepared with different reaction times. Numerous positive-mode TOF mass spectra were acquired from the raster-scanned sample spots of the AChE assays to analyze the progress of the enzyme reaction and to perform mass spectrometry imaging of the sample spots. The reaction kinetics plots obtained by DIUTHAME were found to reflect the time course of reaction progression as much as those obtained by MALDI and were found to have less error variation than MALDI. DIUTHAME allowed the already irradiated sample spots to be reused to reproduce the reaction kinetics plots by the second measurement conducted a week after, whereas MALDI was not able to properly reproduce the kinetics plots by remeasuring the irradiated sample spots. In DIUTHAME, which does not require a matrix, the experimental procedure for measuring AChE assay becomes extremely simple. DIUTHAME is potentially able to provide more precise AChE reaction kinetics plots than what have been accomplished by MALDI.

中文翻译：

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在 DIUTHAME-MS 中监测乙酰胆碱酯酶反应测定的可行性。

DIUTHAME（使用通孔氧化铝膜的解吸电离）是一种新型的无基质激光解吸/电离方法，可实现高重现性的质谱采集。本研究旨在评估 DIUTHAME 对药物发现过程中高通量筛选 (HTS) 常用的乙酰胆碱酯酶反应测定 (AChE 测定) 的适用性。将市售的 9 通道 DIUTHAME 芯片应用于一系列不同反应时间制备的 AChE 检测。从 AChE 检测的光栅扫描样品点获得了大量正模式 TOF 质谱，以分析酶反应的进程并执行样品点的质谱成像。发现由 DIUTHAME 获得的反应动力学图与 MALDI 获得的反应动力学图反映了反应进程的时间过程，并且发现误差变化比 MALDI 小。通过一周后进行的第二次测量，DIUTHAME 允许重复使用已经照射过的样品点来再现反应动力学图，而 MALDI 无法通过重新测量照射过的样品点来正确再现动力学图。在不需要基质的 DIUTHAME 中，测量 AChE 分析的实验程序变得非常简单。DIUTHAME 有可能提供比 MALDI 已经完成的更精确的 AChE 反应动力学图。通过一周后进行的第二次测量，DIUTHAME 允许重复使用已经照射过的样品点来再现反应动力学图，而 MALDI 无法通过重新测量照射过的样品点来正确再现动力学图。在不需要基质的 DIUTHAME 中，测量 AChE 分析的实验程序变得非常简单。DIUTHAME 有可能提供比 MALDI 已经完成的更精确的 AChE 反应动力学图。通过一周后进行的第二次测量，DIUTHAME 允许重复使用已经照射过的样品点来再现反应动力学图，而 MALDI 无法通过重新测量照射过的样品点来正确再现动力学图。在不需要基质的 DIUTHAME 中，测量 AChE 分析的实验程序变得非常简单。DIUTHAME 有可能提供比 MALDI 已经完成的更精确的 AChE 反应动力学图。