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Sphingolipid Analysis by High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS)

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Sphingolipids as Signaling and Regulatory Molecules

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 688))

Abstract

Sphingolipid (SPL) metabolism (Fig. 1) serves a key role in the complex mechanisms regulating cellular stress responses to environment. Several SPL metabolites, especially ceramide (Cer), sphingosine (Sph) and sphingosine1-phosphate (S1P) act as key bioactive molecules governing cell growth and programmed cell death (Fig. 2). Perturbations in sphingolipids of one type may enhance or interfere with the action of another. To monitor changes in SPL composition therefore, reliable analytical methods are necessary.

Here we present the liquid chromatography tandem mass spectrometry (LC-MS/MS) approach for simultaneous qualitative and quantitative monitoring of SPL components (classes and molecular species) in biological material as an effective tool to study sphingolipid signaling events. The LC-MS/MS methodology is the only available technique that provides high specificity and sensitivity, along with a wealth of structural identification information.

Sphingolipid biosynthesis and metabolic pathways; metabolomic profiling of sphingolipids. Abbreviations used in the figure: 3-keto-dhSph, 3-keto-dihydrosphingosine; dhSph, dihydrosphingosine; dhS1P, dihydrosphingosine 1-phosphate; dhCer, dihydroceramide; Sph, sphingosine; S1P, sphingosine 1-phosphate; Cer1P, ceramide-1-phosphate; SM, sphingomyelin; lyso-SM, lyso-sphingomyelin; DAG, diacylglycerol; O-Acyl-Sph. O-acyl-sphingosine, O-Acyl-Cer, O-acyl-ceramide; N-Me-Sph, N-methyl-sphingosine; N.N-DMSph, N,N-dimethyl-sphingosine; PA, palmitic acid; EAP, ethanolamine phosphate; HD, hexadecenal; GL, glycerolipids; de-SM, demethylated sphingomyelin; PGLs, phosphoglycerolipids; GlcCer, glucosylceramide; GalCer, galactosylceramide; GSLs, glycosphingolipids.

Natural sphingolipids are a highly heterogenous system related to the sphingoid bases and derivatization made on the amino- and hydroxy-functions. Structures shown in this figure represent derivatives of 18C-SB (sphingoid bases containing C18-backbone chain) indicating SPLs containing 2-amino-1,3-dihydroxy-octadecene-4E, 2-amino-1,3-dihydroxy-octadecane and 2-amino-1,3,4-trihydroxy-octadecane. General structures, nomenclature and abbreviations for SPLs are cited and described in this presentation. Cn -indicates the chain length of N-acyl part of SPLs.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave., P.O. Box 250509, Charleston, South Carolina, 29425, USA

    Jacek Bielawski, Jason S. Pierce, Justin Snider, Barbara Rembiesa, Zdzislaw M. Szulc & Alicja Bielawska

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  1. Jacek Bielawski
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  2. Jason S. Pierce
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Correspondence to Jacek Bielawski .

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Editors and Affiliations

  1. Department of Biochemistry, Virginia Commonwealth University, Richmond, Virginia, USA

    Charles Chalfant PhD

  2. Department of Biochemistry, Medical University of South Carolina, Charleston, South Carolina, USA

    Maurizio Del Poeta MD

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Bielawski, J., Pierce, J.S., Snider, J., Rembiesa, B., Szulc, Z.M., Bielawska, A. (2010). Sphingolipid Analysis by High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS). In: Chalfant, C., Poeta, M.D. (eds) Sphingolipids as Signaling and Regulatory Molecules. Advances in Experimental Medicine and Biology, vol 688. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-6741-1_3

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