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

6 Citations (Scopus)

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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Sphingolipids
Liquid chromatography
Biomolecules
Sorting
Mass spectrometry
Proteins
Cells
Throughput
Membranes
Monitoring

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

A Comprehensive High-Resolution Targeted Workflow for the Deep Profiling of Sphingolipids. / Peng, Bing; Weintraub, Susan E; Coman, Cristina; Ponnaiyan, Srigayatri; Sharma, Rakesh; Tews, Björn; Winter, Dominic; Ahrends, Robert.

In: Analytical Chemistry, Vol. 89, No. 22, 21.11.2017, p. 12480-12487.

Research output: Contribution to journalArticle

Peng, B, Weintraub, SE, 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, vol. 89, no. 22, pp. 12480-12487. https://doi.org/10.1021/acs.analchem.7b03576
Peng, Bing ; Weintraub, Susan E ; Coman, Cristina ; Ponnaiyan, Srigayatri ; Sharma, Rakesh ; Tews, Björn ; Winter, Dominic ; Ahrends, Robert. / A Comprehensive High-Resolution Targeted Workflow for the Deep Profiling of Sphingolipids. In: Analytical Chemistry. 2017 ; Vol. 89, No. 22. pp. 12480-12487.
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