Comparison of different lipid extraction methods for roots of barley

Autores
Reyna, M.; Vilchez, Ana Carolina; Peppino Margutti, Micaela Yesica; Villasuso, Ana Laura
Año de publicación
2020
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds in solution at concentrations adequate for subsequent detection, quantitation and analysis through spectroscopic methods are required. To date, numerous methods are used to extract lipids from plant tissues. However, a comprehensive analysis of the efficiency and reproducibility of these methods to extract multiple lipid classes from roots of barley has not been undertaken. In this study, we report the comparison of three different lipid extraction procedures in order to determine the most effective lipid extraction protocol to extract lipids from root tissues of the Barley linked to a lipidomicsbased ESI-MS/MS assay coupled with a statistical analysis. While particular methods were best suited to extract different lipid classes from barley roots, overall a single-step extraction method with a 24 h extraction period, which uses a mixture of chloroform, isopropanol, methanol and water, was the most efficient, reproducible and the least labor-intensive to extract a broad range of lipids for untargeted lipidomic analysis of Barley roots. In addition, the optimal protocol for barley root lipid extraction involves immediately submerging plant tissues in hot isopropanol containing 0.01% BHT right upon harvesting of fresh tissues from their respective culturing conditions, which will produce undistorted lipid profiles truly reflective of the endogenous levels of phosphatidylcholine and phosphatidic acid molecular forms (PCs and PAs). Given the critical role of PCs as predominant membrane lipids, as well as the established function of PAs as signal lipid, it is evident that strict adherence to sample pre-treatment protocol is indispensable in deriving biologically meaningful lipid profiles in plants. On the other hands, results indicate that the use of the simplified method allows improving the extraction performance of acidic lipids (PA and phosphatidylserine), which is reflected in a significant increase in the lipid content. Moreover, the data variability obtained by new method extraction was higher than classic method. However, the relation between the amount of extracted lipids and data variability was lower with a single-step extraction method suggesting this extraction could generate a lipid profiles closer to the real composition of the analyzed tissue.
Fil: Reyna, M.. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; Argentina
Fil: Vilchez, Ana Carolina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; Argentina
Fil: Peppino Margutti, Micaela Yesica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Villasuso, Ana Laura. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; Argentina
LVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology y XV Annual Meeting Argentinean Society for General Microbiology
Virtual
Argentina
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Sociedad Argentina de Microbiología General
Materia
Lipids extraction
Barley
Phosphatidic acid
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:11336/271600
Acceder
id CONICETDig_8f5d6f49c9c7325aad255c8a13a5806f
oai_identifier_str oai:ri.conicet.gov.ar:11336/271600
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Comparison of different lipid extraction methods for roots of barleyReyna, M.Vilchez, Ana CarolinaPeppino Margutti, Micaela YesicaVillasuso, Ana LauraLipids extractionBarleyPhosphatidic acidhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds in solution at concentrations adequate for subsequent detection, quantitation and analysis through spectroscopic methods are required. To date, numerous methods are used to extract lipids from plant tissues. However, a comprehensive analysis of the efficiency and reproducibility of these methods to extract multiple lipid classes from roots of barley has not been undertaken. In this study, we report the comparison of three different lipid extraction procedures in order to determine the most effective lipid extraction protocol to extract lipids from root tissues of the Barley linked to a lipidomicsbased ESI-MS/MS assay coupled with a statistical analysis. While particular methods were best suited to extract different lipid classes from barley roots, overall a single-step extraction method with a 24 h extraction period, which uses a mixture of chloroform, isopropanol, methanol and water, was the most efficient, reproducible and the least labor-intensive to extract a broad range of lipids for untargeted lipidomic analysis of Barley roots. In addition, the optimal protocol for barley root lipid extraction involves immediately submerging plant tissues in hot isopropanol containing 0.01% BHT right upon harvesting of fresh tissues from their respective culturing conditions, which will produce undistorted lipid profiles truly reflective of the endogenous levels of phosphatidylcholine and phosphatidic acid molecular forms (PCs and PAs). Given the critical role of PCs as predominant membrane lipids, as well as the established function of PAs as signal lipid, it is evident that strict adherence to sample pre-treatment protocol is indispensable in deriving biologically meaningful lipid profiles in plants. On the other hands, results indicate that the use of the simplified method allows improving the extraction performance of acidic lipids (PA and phosphatidylserine), which is reflected in a significant increase in the lipid content. Moreover, the data variability obtained by new method extraction was higher than classic method. However, the relation between the amount of extracted lipids and data variability was lower with a single-step extraction method suggesting this extraction could generate a lipid profiles closer to the real composition of the analyzed tissue.Fil: Reyna, M.. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; ArgentinaFil: Vilchez, Ana Carolina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; ArgentinaFil: Peppino Margutti, Micaela Yesica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Villasuso, Ana Laura. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; ArgentinaLVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology y XV Annual Meeting Argentinean Society for General MicrobiologyVirtualArgentinaSociedad Argentina de Investigación en Bioquímica y Biología MolecularSociedad Argentina de Microbiología GeneralTech Science Press2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectEncuentroJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttp://hdl.handle.net/11336/271600Comparison of different lipid extraction methods for roots of barley; LVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology y XV Annual Meeting Argentinean Society for General Microbiology; Virtual; Argentina; 2020; 92-930327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://samige.org.ar/libros/2020.pdfinfo:eu-repo/semantics/altIdentifier/url/https://samige.org.ar/congresos-anteriores/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T12:08:03Zoai:ri.conicet.gov.ar:11336/271600instacron: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:34982025-10-22 12:08:04.074CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Comparison of different lipid extraction methods for roots of barley
title Comparison of different lipid extraction methods for roots of barley
spellingShingle Comparison of different lipid extraction methods for roots of barley
Reyna, M.
Lipids extraction
Barley
Phosphatidic acid
title_short Comparison of different lipid extraction methods for roots of barley
title_full Comparison of different lipid extraction methods for roots of barley
title_fullStr Comparison of different lipid extraction methods for roots of barley
title_full_unstemmed Comparison of different lipid extraction methods for roots of barley
title_sort Comparison of different lipid extraction methods for roots of barley
dc.creator.none.fl_str_mv Reyna, M.
Vilchez, Ana Carolina
Peppino Margutti, Micaela Yesica
Villasuso, Ana Laura
author Reyna, M.
author_facet Reyna, M.
Vilchez, Ana Carolina
Peppino Margutti, Micaela Yesica
Villasuso, Ana Laura
author_role author
author2 Vilchez, Ana Carolina
Peppino Margutti, Micaela Yesica
Villasuso, Ana Laura
author2_role author
author
author
dc.subject.none.fl_str_mv Lipids extraction
Barley
Phosphatidic acid
topic Lipids extraction
Barley
Phosphatidic acid
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds in solution at concentrations adequate for subsequent detection, quantitation and analysis through spectroscopic methods are required. To date, numerous methods are used to extract lipids from plant tissues. However, a comprehensive analysis of the efficiency and reproducibility of these methods to extract multiple lipid classes from roots of barley has not been undertaken. In this study, we report the comparison of three different lipid extraction procedures in order to determine the most effective lipid extraction protocol to extract lipids from root tissues of the Barley linked to a lipidomicsbased ESI-MS/MS assay coupled with a statistical analysis. While particular methods were best suited to extract different lipid classes from barley roots, overall a single-step extraction method with a 24 h extraction period, which uses a mixture of chloroform, isopropanol, methanol and water, was the most efficient, reproducible and the least labor-intensive to extract a broad range of lipids for untargeted lipidomic analysis of Barley roots. In addition, the optimal protocol for barley root lipid extraction involves immediately submerging plant tissues in hot isopropanol containing 0.01% BHT right upon harvesting of fresh tissues from their respective culturing conditions, which will produce undistorted lipid profiles truly reflective of the endogenous levels of phosphatidylcholine and phosphatidic acid molecular forms (PCs and PAs). Given the critical role of PCs as predominant membrane lipids, as well as the established function of PAs as signal lipid, it is evident that strict adherence to sample pre-treatment protocol is indispensable in deriving biologically meaningful lipid profiles in plants. On the other hands, results indicate that the use of the simplified method allows improving the extraction performance of acidic lipids (PA and phosphatidylserine), which is reflected in a significant increase in the lipid content. Moreover, the data variability obtained by new method extraction was higher than classic method. However, the relation between the amount of extracted lipids and data variability was lower with a single-step extraction method suggesting this extraction could generate a lipid profiles closer to the real composition of the analyzed tissue.
Fil: Reyna, M.. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; Argentina
Fil: Vilchez, Ana Carolina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; Argentina
Fil: Peppino Margutti, Micaela Yesica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Villasuso, Ana Laura. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Biotecnología Ambiental y Salud - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Biotecnología Ambiental y Salud; Argentina
LVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology y XV Annual Meeting Argentinean Society for General Microbiology
Virtual
Argentina
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Sociedad Argentina de Microbiología General
description The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds in solution at concentrations adequate for subsequent detection, quantitation and analysis through spectroscopic methods are required. To date, numerous methods are used to extract lipids from plant tissues. However, a comprehensive analysis of the efficiency and reproducibility of these methods to extract multiple lipid classes from roots of barley has not been undertaken. In this study, we report the comparison of three different lipid extraction procedures in order to determine the most effective lipid extraction protocol to extract lipids from root tissues of the Barley linked to a lipidomicsbased ESI-MS/MS assay coupled with a statistical analysis. While particular methods were best suited to extract different lipid classes from barley roots, overall a single-step extraction method with a 24 h extraction period, which uses a mixture of chloroform, isopropanol, methanol and water, was the most efficient, reproducible and the least labor-intensive to extract a broad range of lipids for untargeted lipidomic analysis of Barley roots. In addition, the optimal protocol for barley root lipid extraction involves immediately submerging plant tissues in hot isopropanol containing 0.01% BHT right upon harvesting of fresh tissues from their respective culturing conditions, which will produce undistorted lipid profiles truly reflective of the endogenous levels of phosphatidylcholine and phosphatidic acid molecular forms (PCs and PAs). Given the critical role of PCs as predominant membrane lipids, as well as the established function of PAs as signal lipid, it is evident that strict adherence to sample pre-treatment protocol is indispensable in deriving biologically meaningful lipid profiles in plants. On the other hands, results indicate that the use of the simplified method allows improving the extraction performance of acidic lipids (PA and phosphatidylserine), which is reflected in a significant increase in the lipid content. Moreover, the data variability obtained by new method extraction was higher than classic method. However, the relation between the amount of extracted lipids and data variability was lower with a single-step extraction method suggesting this extraction could generate a lipid profiles closer to the real composition of the analyzed tissue.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Encuentro
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/271600
Comparison of different lipid extraction methods for roots of barley; LVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology y XV Annual Meeting Argentinean Society for General Microbiology; Virtual; Argentina; 2020; 92-93
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/271600
identifier_str_mv Comparison of different lipid extraction methods for roots of barley; LVI Annual Meeting Argentine Society for Biochemistry and Molecular Biology y XV Annual Meeting Argentinean Society for General Microbiology; Virtual; Argentina; 2020; 92-93
0327-9545
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://samige.org.ar/libros/2020.pdf
info:eu-repo/semantics/altIdentifier/url/https://samige.org.ar/congresos-anteriores/
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/vnd.openxmlformats-officedocument.wordprocessingml.document
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
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
_version_ 1846782449271439360
score 12.982451

Publicaciones similares