TY  - JOUR
AU  - Peng, Bing
AU  - Weintraub, Susan T
AU  - Coman, Cristina
AU  - Ponnaiyan, Srigayatri
AU  - Sharma, Rakesh
AU  - Tews, Björn
AU  - Winter, Dominic
AU  - Ahrends, Robert
TI  - A Comprehensive High-Resolution Targeted Workflow for the Deep Profiling of Sphingolipids.
JO  - Analytical chemistry
VL  - 89
IS  - 22
SN  - 1520-6882
CY  - Columbus, Ohio
PB  - American Chemical Society
M1  - DKFZ-2017-06276
SP  - 12480 - 12487
PY  - 2017
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:29039908
DO  - DOI:10.1021/acs.analchem.7b03576
UR  - https://inrepo02.dkfz.de/record/131644
ER  -