%0 Journal Article
%A Peng, Bing
%A Weintraub, Susan T
%A Coman, Cristina
%A Ponnaiyan, Srigayatri
%A Sharma, Rakesh
%A Tews, Björn
%A Winter, Dominic
%A Ahrends, Robert
%T A Comprehensive High-Resolution Targeted Workflow for the Deep Profiling of Sphingolipids.
%J Analytical chemistry
%V 89
%N 22
%@ 1520-6882
%C Columbus, Ohio
%I American Chemical Society
%M DKFZ-2017-06276
%P 12480 - 12487
%D 2017
%X 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.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:29039908
%R 10.1021/acs.analchem.7b03576
%U https://inrepo02.dkfz.de/record/131644