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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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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 |
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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 |
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