Abstract
This protocol describes a method to extract total polar glycerol lipids from plant materials, followed by mass spectrometry profiling. Different glycerol lipid classes can be distinguished by their head-groups, which can be profiled automatically and quantitatively by a triple quadrupole mass spectrometry in multiple reaction monitoring (MRM) mode with an autosampler. Comparing with other established methods, such as thin layer chromatography (TLC) separation followed by Gas spectrometry (GC) analysis, this method requires little effort in sample preparation and separation, while the resolution is not limited to general lipid classes but at side chain level. This method was described and used successfully to profile plant lipids changes under freezing stress in Welti et al. (2002).
Keywords: Mass spectrometry, Lipidomics, Phospholipids, Glycolipids
Materials and Reagents
Equipment
Software
Procedure
Representative data
Figure 1. Illustration of lipid extraction workflow. 1,000 rpm (~200 x g). Figure 2. TLC separation of phospholipids. Lipids extracted in section A can be resolved on a TLC plate. The mobile phase used here is chloroform:methanol:acetic acid:H2O = 85:15:12.5:3.5 (v/v). Figure 3. Representative MS data. A. Different lipid classes were profiled in tandem periods. Period 1 is for control purpose and not used in calculation. B. PE profile in IS sample. C. PE profile in plant sample. D. Enlarged view of plant PE species.
Notes
Recipes
Acknowledgments
This protocol was adapted from Welti et al. (2002). Dr. Welti also provided help in lipid standards, data processing and analysis. Work was supported by the U.S. Department of Energy (DOE) Grant DE-AR0000202 and by the National Science Foundation Grants MCB-1412901 and DBI-1427621.
References
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