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Screening of acetylcholinesterase inhibitors by CE after enzymatic reaction at capillary inlet by Yolanda Martín‐Biosca; Lucia Asensi‐Bernardi; Rosa M. Villanueva‐Camañas; Salvador Sagrado; Maria J. Medina‐Hernández is a Chemistry article available to read on EtoBox.

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## Abstract In this study the development of a procedure based on capillary electrophoresis after enzymatic reaction at capillary inlet methodology for the screening and __in vitro__ evaluation of the biological activity of acetylcholinesterase (AChE) inhibitors is presented. The progress of the enzymatic reaction of the hydrolysis of acetylthiocholine at pH 8 in the presence of AChE and the inhibitor studied is determined by measuring at 230 nm the peak area of the reaction product thiocholine (TCh). In the method employed the capillary was first filled with 30 mM borate‐phosphate buffer (pH 8.0) and subsequently, plugs of: (i) water, (ii) AChE solution, (iii) substrate solution with or without inhibitor, (iv) AChE solution, and (v) water, were hydrodynamically injected into the capillary, and were allowed to stand (and react) during a waiting period of 2 min. The applicability of the proposed methodology to estimate different kinetic parameters of interest such as inhibition constants __K__~i~, identification of inhibitory action mechanism and IC~50~, is evaluated using compounds with known activity, tacrine edrophonium, and neostigmine. The results obtained are compared with bib

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Author

Yolanda Martín‐Biosca; Lucia Asensi‐Bernardi; Rosa M. Villanueva‐Camañas; Salvador Sagrado; Maria J. Medina‐Hernández

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley (ISSN 1615-9306)

Published

2009

Language

EN

Field

Chemistry (Physical Sciences)