A Comprehensive High-Resolution Targeted Workflow for the Deep Profiling of Sphingolipids

Bing Peng, Susan E Weintraub, Cristina Coman, Srigayatri Ponnaiyan, Rakesh Sharma, Björn Tews, Dominic Winter, Robert Ahrends

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Sphingolipids make up a highly diverse group of biomolecules that not only are membrane components but also are involved in various cellular functions such as signaling and protein sorting. To obtain a quantitative view of the sphingolipidome, sensitive, accurate, and comprehensive methods are needed. Here, we present a targeted reversed-phase liquid chromatography-high-resolution mass spectrometry-based workflow that significantly increases the accuracy of measured sphingolipids by resolving nearly isobaric and isobaric species; this is accomplished by a use of (i) an optimized extraction procedure, (ii) a segmented gradient, and (iii) parallel reaction monitoring of a sphingolipid specific fragmentation pattern. The workflow was benchmarked against an accepted sphingolipid model system, the RAW 264.7 cell line, and 61 sphingolipids were quantified over a dynamic range of 7 orders of magnitude, with detection limits in the low femtomole per milligram of protein level, making this workflow an extremely versatile tool for high-throughput sphingolipidomics.

Original languageEnglish (US)
Pages (from-to)12480-12487
Number of pages8
JournalAnalytical Chemistry
Volume89
Issue number22
DOIs
StatePublished - Nov 21 2017

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ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Peng, B., Weintraub, S. E., Coman, C., Ponnaiyan, S., Sharma, R., Tews, B., Winter, D., & Ahrends, R. (2017). A Comprehensive High-Resolution Targeted Workflow for the Deep Profiling of Sphingolipids. Analytical Chemistry, 89(22), 12480-12487. https://doi.org/10.1021/acs.analchem.7b03576