Acetylcholinesterase inhibitors are currently the first choice for the clinical treatment of AD.With the improvement of science and technology,the screening methods are also in constant innovation.This article introduced the research progress of AD therapeutic targets and drug research based on the"cholinergic damage theory",and then emphatically expounded the current methods that could be used for acetylcholinesterase inhibitors’screening,which including Ellman's colorimetric method based on ultraviolet spectrophotometer,thin layer biological autoradiography method,fluorescence analysis method,and ultra-high performance liquid chromatography-mass spectrometry combined with their principles and advantages or disadvantages.We hoped that it would be more effectively and quickly to screen new acetylcholinesterase inhibitors from natural products by summarizing the experience and keeping innovating.