Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors

Autores
Ramallo, Ivana Ayelen; Salazar, Mario Oscar; Furlan, Ricardo Luis Eugenio
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Introduction: The prevailing treatment for Alzheimer's disease is the use of acetylcholinesterase (AChE) inhibitors. Natural extracts are 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 mass spectrometry (HRMS) and thin layer chromatography (TLC)-biological staining. Methodology: The strategy uses an autographic assay based on the α-naphthyl acetate – fast blue B system for the detection of AChE activity. The immobilisation of AChE in agar allowed the extraction of the compounds for analysis by HRMS. Three TLC experiments employing different solvent systems were used in parallel and the mass spectra of the compounds extracted from the 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. spiked with the inhibitor. Similarly, caffeine was straightforwardly spotted as responsible for the inhibitory properties of an extract of Ilex paraguariensis Saint-Hilaire. Comparison of the HRMS profiles lead to the facile identification of the [M+H]+ and [M+Na]+ of the compounds responsible for the inhibition. Conclusion: The proposed methodology, coupling TLC-AChE autography-HRMS, illustrates the feasibility of assigning molecular formulas of active compounds present in complex mixtures directly from autography. The new AChE agar-immobilised assay presented a more homogenous colour and a better definition than direct spraying methods, reducing the cost of the assay and improving its sensitivity.
Fil: Ramallo, Ivana Ayelen. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Salazar, Mario Oscar. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Furlan, Ricardo Luis Eugenio. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Materia
Acetylcholinesterase
Bioautography
Hrms
Thin Layer Chromatography
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/13392
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/13392
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitorsRamallo, Ivana AyelenSalazar, Mario OscarFurlan, Ricardo Luis EugenioAcetylcholinesteraseBioautographyHrmsThin Layer Chromatographyhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Introduction: The prevailing treatment for Alzheimer's disease is the use of acetylcholinesterase (AChE) inhibitors. Natural extracts are 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 mass spectrometry (HRMS) and thin layer chromatography (TLC)-biological staining. Methodology: The strategy uses an autographic assay based on the α-naphthyl acetate – fast blue B system for the detection of AChE activity. The immobilisation of AChE in agar allowed the extraction of the compounds for analysis by HRMS. Three TLC experiments employing different solvent systems were used in parallel and the mass spectra of the compounds extracted from the 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. spiked with the inhibitor. Similarly, caffeine was straightforwardly spotted as responsible for the inhibitory properties of an extract of Ilex paraguariensis Saint-Hilaire. Comparison of the HRMS profiles lead to the facile identification of the [M+H]+ and [M+Na]+ of the compounds responsible for the inhibition. Conclusion: The proposed methodology, coupling TLC-AChE autography-HRMS, illustrates the feasibility of assigning molecular formulas of active compounds present in complex mixtures directly from autography. The new AChE agar-immobilised assay presented a more homogenous colour and a better definition than direct spraying methods, reducing the cost of the assay and improving its sensitivity.Fil: Ramallo, Ivana Ayelen. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Salazar, Mario Oscar. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Furlan, Ricardo Luis Eugenio. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; ArgentinaWiley2015-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/13392Ramallo, Ivana Ayelen; Salazar, Mario Oscar; Furlan, Ricardo Luis Eugenio; Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors; Wiley; Phytochemical Analysis; 26; 6; 11-2015; 404-4121099-1565enginfo:eu-repo/semantics/altIdentifier/doi/10.1002/pca.2574info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/pca.2574/abstractinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:10:07Zoai:ri.conicet.gov.ar:11336/13392instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:10:08.139CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
title Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
spellingShingle Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
Ramallo, Ivana Ayelen
Acetylcholinesterase
Bioautography
Hrms
Thin Layer Chromatography
title_short Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
title_full Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
title_fullStr Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
title_full_unstemmed Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
title_sort Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors
dc.creator.none.fl_str_mv Ramallo, Ivana Ayelen
Salazar, Mario Oscar
Furlan, Ricardo Luis Eugenio
author Ramallo, Ivana Ayelen
author_facet Ramallo, Ivana Ayelen
Salazar, Mario Oscar
Furlan, Ricardo Luis Eugenio
author_role author
author2 Salazar, Mario Oscar
Furlan, Ricardo Luis Eugenio
author2_role author
author
dc.subject.none.fl_str_mv Acetylcholinesterase
Bioautography
Hrms
Thin Layer Chromatography
topic Acetylcholinesterase
Bioautography
Hrms
Thin Layer Chromatography
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Introduction: The prevailing treatment for Alzheimer's disease is the use of acetylcholinesterase (AChE) inhibitors. Natural extracts are 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 mass spectrometry (HRMS) and thin layer chromatography (TLC)-biological staining. Methodology: The strategy uses an autographic assay based on the α-naphthyl acetate – fast blue B system for the detection of AChE activity. The immobilisation of AChE in agar allowed the extraction of the compounds for analysis by HRMS. Three TLC experiments employing different solvent systems were used in parallel and the mass spectra of the compounds extracted from the 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. spiked with the inhibitor. Similarly, caffeine was straightforwardly spotted as responsible for the inhibitory properties of an extract of Ilex paraguariensis Saint-Hilaire. Comparison of the HRMS profiles lead to the facile identification of the [M+H]+ and [M+Na]+ of the compounds responsible for the inhibition. Conclusion: The proposed methodology, coupling TLC-AChE autography-HRMS, illustrates the feasibility of assigning molecular formulas of active compounds present in complex mixtures directly from autography. The new AChE agar-immobilised assay presented a more homogenous colour and a better definition than direct spraying methods, reducing the cost of the assay and improving its sensitivity.
Fil: Ramallo, Ivana Ayelen. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Salazar, Mario Oscar. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Furlan, Ricardo Luis Eugenio. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Inv. Para El Descubrimiento de Farmacos de Rosario; Argentina. Universidad Nacional de Rosario; Argentina
description Introduction: The prevailing treatment for Alzheimer's disease is the use of acetylcholinesterase (AChE) inhibitors. Natural extracts are 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 mass spectrometry (HRMS) and thin layer chromatography (TLC)-biological staining. Methodology: The strategy uses an autographic assay based on the α-naphthyl acetate – fast blue B system for the detection of AChE activity. The immobilisation of AChE in agar allowed the extraction of the compounds for analysis by HRMS. Three TLC experiments employing different solvent systems were used in parallel and the mass spectra of the compounds extracted from the 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. spiked with the inhibitor. Similarly, caffeine was straightforwardly spotted as responsible for the inhibitory properties of an extract of Ilex paraguariensis Saint-Hilaire. Comparison of the HRMS profiles lead to the facile identification of the [M+H]+ and [M+Na]+ of the compounds responsible for the inhibition. Conclusion: The proposed methodology, coupling TLC-AChE autography-HRMS, illustrates the feasibility of assigning molecular formulas of active compounds present in complex mixtures directly from autography. The new AChE agar-immobilised assay presented a more homogenous colour and a better definition than direct spraying methods, reducing the cost of the assay and improving its sensitivity.
publishDate 2015
dc.date.none.fl_str_mv 2015-11
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/13392
Ramallo, Ivana Ayelen; Salazar, Mario Oscar; Furlan, Ricardo Luis Eugenio; Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors; Wiley; Phytochemical Analysis; 26; 6; 11-2015; 404-412
1099-1565
url http://hdl.handle.net/11336/13392
identifier_str_mv Ramallo, Ivana Ayelen; Salazar, Mario Oscar; Furlan, Ricardo Luis Eugenio; Thin layer Chromatography-Autography-High resolution mass spectrometry analysis: accelerating the identification of Acetylcholinesterase inhibitors; Wiley; Phytochemical Analysis; 26; 6; 11-2015; 404-412
1099-1565
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1002/pca.2574
info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/pca.2574/abstract
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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