Planta Med 2012; 78 - PJ62
DOI: 10.1055/s-0032-1321222

Normal phase HPLC profiling of the acetylcholinesterase activity in apolar plant extracts

A Landreau 1, S Bertrand 2, C Simoes-Pires 2, L Marcourt 2, TD Bach 1, M Litaudon 3, D Guilet 1, P Richomme 1, PA Carrupt 2, JL Wolfender 2
  • 1PRES LUNAM, Université d'Angers, Laboratoire SONAS-UFR Sciences Pharmaceutiques et Ingénierie de la Santé, Angers, France
  • 2School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, Geneva, Switzerland
  • 3Centre de Recherche de Gif, Institut de Chimie des Substances Naturelles, CNRS, Gif-sur-Yvette Cedex, France

Among nineteen evaluated Clusiaceous species, one stem bark CH2Cl2 crude extract was selected based on a significant inhibition of acetylcholinesterase (AChE) using the micro-dilution Ellman's method [1]. A normal phase HPLC profiling with micro-fractionation of this extract provided discrete fractions every 20 seconds. In order to obtain a comprehensive profiling of AChE activity all microfractions were tested [2] in dilution assay (Ellman) as well as by bioautography (the Fast Blue B salt method). Furthermore the potency of inhibition was evaluated both by keeping the genuine concentration within the extract and after normalisation to a standard concentration level. From the active fractions five pure compounds were isolated and identified. The different methods of sample preparation and biological evaluation associated with normal-phase micro-fractionation of plant extracts are critically discussed.

[1] Brühlmann et al., Chem. Biodivers., 2004, 1, 819; [2] Wolfender et al., Chimia, 2011, 65, 400