Pharmaceutical Sciences Faculty Publications

Quantitation of Ceramide Phosphorylethanolamines Containing Saturated and Unsaturated Sphingoid Base Cores

Document Type

Article

Publication Date

5-15-2010

Journal Title

Analytical Biochemistry

ISSN

1096-0309

Volume

400

Issue

2

First Page

259

Last Page

269

DOI

10.1016/j.ab.2010.01.033

PubMed ID

20122889

PubMed Central® ID

PMC3020903

Abstract

Sphingomyelin (SM) and ceramide-phosphoethanolamines (cer-PEs) are related lipids present in mammals and insects, respectively. Owing to the critical roles that cer-PEs play in eukaryotic cellular function, there is a need to develop methods that provide accurate quantitation of these compounds. Results obtained in this study demonstrate that Drosophila contains cer-PEs with unsaturated sphingoid base cores as well as low levels of cer-PEs that possess saturated sphingoid base cores. Specifically, the method developed in this study enabled the quantitation of picogram amounts of cer-PE containing both unsaturated d14:1(Delta4) and d16:1(Delta4) and saturated d14:0 sphingoid base cores. Using this method, cer-PE compounds with both saturated and unsaturated sphingoid base cores were initially identified by neutral loss scanning, followed by quantitation using selected reaction monitoring (SRM) scans. The SRM scans measured a product ion originating from the sphingoid base backbone, rather than from the head group, increasing the specificity and sensitivity of the quantitation measurement.

Keywords

Chromatography, high pressure liquid, reverse-phase, drosophila, spectrometry, mass, electrospray ionization, sphingomyelins

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