Cargando…

An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid

BACKGROUND: Lipidomics plays an important role in understanding plant adaptation to different stresses and improving our knowledge of the genes underlying lipid metabolism. Lipidomics involves lipid extraction, sample preparation, mass spectrometry analysis, and data interpretation. One of the pract...

Descripción completa

Detalles Bibliográficos
Autores principales: Shiva, Sunitha, Enninful, Regina, Roth, Mary R., Tamura, Pamela, Jagadish, Krishna, Welti, Ruth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5812192/
https://www.ncbi.nlm.nih.gov/pubmed/29449874
http://dx.doi.org/10.1186/s13007-018-0282-y
_version_ 1783299999775326208
author Shiva, Sunitha
Enninful, Regina
Roth, Mary R.
Tamura, Pamela
Jagadish, Krishna
Welti, Ruth
author_facet Shiva, Sunitha
Enninful, Regina
Roth, Mary R.
Tamura, Pamela
Jagadish, Krishna
Welti, Ruth
author_sort Shiva, Sunitha
collection PubMed
description BACKGROUND: Lipidomics plays an important role in understanding plant adaptation to different stresses and improving our knowledge of the genes underlying lipid metabolism. Lipidomics involves lipid extraction, sample preparation, mass spectrometry analysis, and data interpretation. One of the practical challenges for large-scale lipidomics studies on plant leaves is the requirement of an efficient and rapid extraction method. RESULTS: A single-extraction method with a polar solvent mixture gives results comparable to a widely used, multi-extraction method when tested on both Arabidopsis thaliana and Sorghum bicolor leaf tissue. This single-extraction method uses a mixture of 30 parts chloroform, 25 parts isopropanol, 41.5 parts methanol, and 3.5 parts water (v/v/v/v) and a 24-h extraction time. Neither inclusion of ammonium acetate nor inclusion of acetic acid increased extraction efficiency. CONCLUSIONS: The extract produced by this method can be used for analysis by mass spectrometry without a solvent evaporation step. The amount of lipid extracted, including phosphatidic acid, is comparable to widely used, more labor-intensive methods. The single-extraction protocol is less laborious, reducing the potential for human error. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-018-0282-y) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5812192
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-58121922018-02-15 An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid Shiva, Sunitha Enninful, Regina Roth, Mary R. Tamura, Pamela Jagadish, Krishna Welti, Ruth Plant Methods Methodology BACKGROUND: Lipidomics plays an important role in understanding plant adaptation to different stresses and improving our knowledge of the genes underlying lipid metabolism. Lipidomics involves lipid extraction, sample preparation, mass spectrometry analysis, and data interpretation. One of the practical challenges for large-scale lipidomics studies on plant leaves is the requirement of an efficient and rapid extraction method. RESULTS: A single-extraction method with a polar solvent mixture gives results comparable to a widely used, multi-extraction method when tested on both Arabidopsis thaliana and Sorghum bicolor leaf tissue. This single-extraction method uses a mixture of 30 parts chloroform, 25 parts isopropanol, 41.5 parts methanol, and 3.5 parts water (v/v/v/v) and a 24-h extraction time. Neither inclusion of ammonium acetate nor inclusion of acetic acid increased extraction efficiency. CONCLUSIONS: The extract produced by this method can be used for analysis by mass spectrometry without a solvent evaporation step. The amount of lipid extracted, including phosphatidic acid, is comparable to widely used, more labor-intensive methods. The single-extraction protocol is less laborious, reducing the potential for human error. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-018-0282-y) contains supplementary material, which is available to authorized users. BioMed Central 2018-02-13 /pmc/articles/PMC5812192/ /pubmed/29449874 http://dx.doi.org/10.1186/s13007-018-0282-y Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Shiva, Sunitha
Enninful, Regina
Roth, Mary R.
Tamura, Pamela
Jagadish, Krishna
Welti, Ruth
An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid
title An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid
title_full An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid
title_fullStr An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid
title_full_unstemmed An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid
title_short An efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid
title_sort efficient modified method for plant leaf lipid extraction results in improved recovery of phosphatidic acid
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5812192/
https://www.ncbi.nlm.nih.gov/pubmed/29449874
http://dx.doi.org/10.1186/s13007-018-0282-y
work_keys_str_mv AT shivasunitha anefficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT enninfulregina anefficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT rothmaryr anefficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT tamurapamela anefficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT jagadishkrishna anefficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT weltiruth anefficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT shivasunitha efficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT enninfulregina efficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT rothmaryr efficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT tamurapamela efficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT jagadishkrishna efficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid
AT weltiruth efficientmodifiedmethodforplantleaflipidextractionresultsinimprovedrecoveryofphosphatidicacid