Abstract
Sphingolipids generate signals in plants in response to a variety of biotic and abiotic stresses. Measuring these signaling compounds is complicated by the heterogeneity of structures within the sphingolipid family and the comparatively low concentration of their metabolites in plant tissues. To date, the only method with the sensitivity, dynamic range, and specificity to measure all sphingolipids in a plant extract is liquid chromatography coupled to mass spectrometry. The drawback of this method is the cost of the hardware, the expertise in mass spectrometry required to critically assess the outcome and the lack of suitable standards for accurate quantitative analysis. The goal of this chapter is to assist researchers in setting up experiments to measure sphingolipids and explain some of the pitfalls and solutions along the way.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Lester RL, Dickson RC (2001) High-performance liquid chromatography analysis of molecular species of sphingolipid-related long chain bases and long chain base phosphates in Saccharomyces cerevisiae after derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Anal Biochem 298:283–292
Merrill AH Jr, Caligan TB, Wang E et al (2000) Analysis of sphingoid bases and sphingoid base 1-phosphates by high-performance liquid chromatography. Methods Enzymol 312:3–9
Cahoon EB, Lynch DV (1991) Analysis of glucocerebrosides of rye (Secale cereale L. cv Puma) leaf and plasma membrane. Plant Physiol 95:58–68
Imai H, Morimoto Y, Tamura K (2000) Sphingoid base composition of monoglucosylceramide in Brassicaceae. J Plant Physiol 157:453–456
Sullards MC (2000) Analysis of sphingomyelin, glucosylceramide, ceramide, sphingosine, and sphingosine 1-phosphate by tandem mass spectrometry. Methods Enzymol 312:32–45
Markham JE, Jaworski JG (2007) Rapid measurement of sphingolipids from Arabidopsis thaliana by reversed-phase high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. Rapid Commun Mass Spectrom 21:1304–1314
Chen M, Markham JE, Dietrich CR et al (2008) Sphingolipid long-chain base hydroxylation is important for growth and regulation of sphingolipid content and composition in Arabidopsis. Plant Cell 20:1862–1878
Acknowledgments
This work was supported by NSF grant MCB-0843312 and the University of Nebraska-Lincoln.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Markham, J.E. (2013). Detection and Quantification of Plant Sphingolipids by LC-MS. In: Munnik, T., Heilmann, I. (eds) Plant Lipid Signaling Protocols. Methods in Molecular Biology, vol 1009. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-401-2_10
Download citation
DOI: https://doi.org/10.1007/978-1-62703-401-2_10
Published:
Publisher Name: Humana, Totowa, NJ
Print ISBN: 978-1-62703-400-5
Online ISBN: 978-1-62703-401-2
eBook Packages: Springer Protocols