INVESTIGADORES
SALAZAR Mario Oscar
artículos
Título:
Thin Layer Chromatography-Autography-High Resolution Mass Spectrometry Analysis: Accelerating the Identification of Acetylcholinesterase Inhibitors
Autor/es:
I. AYELEN RAMALLO; MARIO O. SALAZAR; RICARDO L. E. FURLAN
Revista:
PHYTOCHEMICAL ANALYSIS
Editorial:
JOHN WILEY & SONS LTD
Referencias:
Lugar: LOndres; Año: 2015
ISSN:
0958-0344
Resumen:
Introduction ? The prevailing treatment for Alzheimer?s disease is the use of acetylcholinesterase (AChE) inhibitors. Natural extractsare the principal source of AChE?s inhibitors. However, their chemical complexity demands for simple, selective and rapid assays.Objective ? To develop a strategy for identification of AChE inhibitors present in mixtures employing high resolution massspectrometry (HRMS) and thin layer chromatography (TLC)-biological staining.Methodology ? The strategy uses an autographic assay based on the á-naphthyl acetate ? fast blue B systemfor the detection ofAChE activity. The immobilisation of AChE in agar allowed the extraction of the compounds for analysis by HRMS. Three TLCexperiments employing different solvent systems were used in parallel and the mass spectra of the compounds extracted fromthe inhibition halos, were compared. The analysis was performed under MatLab environment.Results ? The strategy was used to detect the presence of physostigmine in an extract of Brassica rapa L. spikedwith the inhibitor.Similarly, caffeine was straightforwardly spotted as responsible for the inhibitory properties of an extract of Ilex paraguariensisSaint-Hilaire. Comparison of the HRMS profiles lead to the facile identification of the [M+H]+ and [M+Na]+ of the compoundsresponsible for the inhibition.Conclusion ? The proposedmethodology, coupling TLC-AChE autography-HRMS, illustrates the feasibility of assigningmolecularformulas of active compounds present in complex mixtures directly from autography. The new AChE agar-immobilised assaypresented a more homogenous colour and a better definition than direct spraying methods, reducing the cost of the assayand improving its sensitivity. Copyright © 2015 John Wiley & Sons, Ltd.
