Format:
8
ISSN:
1520-6882
Content:
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.
Note:
Gesehen am 11.07.2018
In:
Analytical chemistry, Columbus, Ohio : American Chemical Society, 1947, 89(2017), 22, Seite 12480-12487, 1520-6882
In:
volume:89
In:
year:2017
In:
number:22
In:
pages:12480-12487
In:
extent:8
Language:
English
DOI:
10.1021/acs.analchem.7b03576
