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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 2018 v.14 no.1 pp. 14
Arabidopsis thaliana, Sorghum bicolor, acetic acid, ammonium acetate, chloroform, evaporation, genes, isopropyl alcohol, leaves, lipid metabolism, lipids, mass spectrometry, methanol, plant adaptation, solvents
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.