Improved Thin-layer Chromatography Bioautographic Assay for the Detection of Actylcholinesterase Inhibitors in Plants

doi:10.1002/pca.1310

	Improved Thin-layer Chromatography Bioautographic Assay for the Detection of Actylcholinesterase Inhibitors in Plants
	Yang, Zhong-Duo 1,2; Song, Zhu-Wen 1; Ren, Jin 1; Yang, Ming-Jun 1; Li, Shuo 1
刊名	PHYTOCHEMICAL ANALYSIS
	2011
卷号	22 期号:6 页码:509-515
关键词	TLC bioautography method Acetylcholinesterase inhibitors new synthesised substrate Alzheimer's disease
ISSN号	0958-0344
DOI	10.1002/pca.1310
英文摘要	Introduction - Thin-layer chromatography (TLC) bioautographic method is a simple and rapid method to screen acetylcholinesterase inhibitors from plant extracts. However, the high consumption of enzyme (6 U/mL) in current methods makes the procedure expensive, which is an obstacle to scientific research centers lacking funding. Objective - To develop a new low-cost TLC bioautographic method. Methodology - A series of compounds, as substrates, were synthesised and their ability to be hydrolysed by acetylcholinesterase was evaluated by the HPLC method. Results - 4-Methoxyphenyl acetate (14) was proved to be an appropriate substrate for TLC bioautographic assay. Therefore a new and cheap TLC bioautographic assay was set up. The mechanism of this new method is that the enzyme converts 4-methoxylphenyl acetate into 4-methoxyphenol, which reacts with a solution of potassium ferricyanide ([K(3)(FeCN)(6)]) and iron chloride hexahydrate (FeCl(3)center dot 6H(2)O) to make an aquamarine blue coloured background on the TLC plates. Regions of the TLC plate which contain acetylcholinesterase inhibitors show up as light yellow spots against the background. The consumption of enzyme (1 U/mL) in the new method is low and the detection limit of two known acetylcholinesterase inhibitors, huperzine A (0.0001 mu g) and physostigmine (0.001 mu g), for this assay are close to published values. Conclusion - A low-cost TLC bioautographic method was developed, which will benefit research groups pursuing natural acetylcholinesterase inhibitors. Copyright (C) 2011 John Wiley & Sons, Ltd.
WOS研究方向	Biochemistry & Molecular Biology ; Plant Sciences ; Chemistry
语种	英语
出版者	WILEY-BLACKWELL
WOS记录号	WOS:000296421100005
内容类型	期刊论文
源URL	[http://ir.lut.edu.cn/handle/2XXMBERH/155551]
专题	生命科学与工程学院
作者单位	1.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou 730050, Peoples R China; 2.Lanzhou Univ, State Key Lab Appl Organ Chem, Lanzhou 730000, Peoples R China
推荐引用方式 GB/T 7714	Yang, Zhong-Duo,Song, Zhu-Wen,Ren, Jin,et al. Improved Thin-layer Chromatography Bioautographic Assay for the Detection of Actylcholinesterase Inhibitors in Plants[J]. PHYTOCHEMICAL ANALYSIS,2011,22(6):509-515.
APA	Yang, Zhong-Duo,Song, Zhu-Wen,Ren, Jin,Yang, Ming-Jun,&Li, Shuo.(2011).Improved Thin-layer Chromatography Bioautographic Assay for the Detection of Actylcholinesterase Inhibitors in Plants.PHYTOCHEMICAL ANALYSIS,22(6),509-515.
MLA	Yang, Zhong-Duo,et al."Improved Thin-layer Chromatography Bioautographic Assay for the Detection of Actylcholinesterase Inhibitors in Plants".PHYTOCHEMICAL ANALYSIS 22.6(2011):509-515.