A comprehensive high-resolution targeted workflow for the deep profiling of sphingolipids

• Verfasst von: Peng, Bing [VerfasserIn]
• Verfasst von: Sharma, Rakesh [VerfasserIn]
• Verfasst von: Tews, Björn [VerfasserIn]
• Titel: A comprehensive high-resolution targeted workflow for the deep profiling of sphingolipids
• Verf.angabe: Bing Peng, Susan T. Weintraub, Cristina Coman, Srigayatri Ponnaiyan, Rakesh Sharma, Björn Tews, Dominic Winter, and Robert Ahrends
• E-Jahr: 2017
• Jahr: October 17, 2017
• Umfang: 8 S.
• Fussnoten: Gesehen am 11.07.2018
• Titel Quelle: Enthalten in: Analytical chemistry
• Ort Quelle: Columbus, Ohio : American Chemical Society, 1947
• Jahr Quelle: 2017
• Band/Heft Quelle: 89(2017), 22, Seite 12480-12487
• ISSN Quelle: 1520-6882
• DOI: doi:10.1021/acs.analchem.7b03576
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1577521293

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.