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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/271600
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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 Técnicas2025-09-29T10:35:41Zoai: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:34982025-09-29 10:35:42.015CONICET 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 |
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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 |
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publishedVersion |
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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 |
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eng |
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eng |
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Tech Science Press |
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Tech Science Press |
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