A comprehensive high-resolution targeted workflow for the deep profiling of sphingolipids
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 neede...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
October 17, 2017
|
| In: |
Analytical chemistry
Year: 2017, Volume: 89, Issue: 22, Pages: 12480-12487 |
| ISSN: | 1520-6882 |
| DOI: | 10.1021/acs.analchem.7b03576 |
| Online Access: | Verlag, Volltext: http://dx.doi.org/10.1021/acs.analchem.7b03576 Verlag, Volltext: https://doi.org/10.1021/acs.analchem.7b03576 
|
| Author Notes: | Bing Peng, Susan T. Weintraub, Cristina Coman, Srigayatri Ponnaiyan, Rakesh Sharma, Björn Tews, Dominic Winter, and Robert Ahrends |
| Summary: | 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. |
|---|---|
| Item Description: | Gesehen am 11.07.2018 |
| Physical Description: | Online Resource |
| ISSN: | 1520-6882 |
| DOI: | 10.1021/acs.analchem.7b03576 |