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000131644 037__ $$aDKFZ-2017-06276
000131644 041__ $$aeng
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000131644 1001_ $$aPeng, Bing$$b0
000131644 245__ $$aA Comprehensive High-Resolution Targeted Workflow for the Deep Profiling of Sphingolipids.
000131644 260__ $$aColumbus, Ohio$$bAmerican Chemical Society$$c2017
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000131644 520__ $$aSphingolipids make up a highly diverse group of biomolecules that not only are membrane components but also are involved in various cellular functions such as signaling and protein sorting. To obtain a quantitative view of the sphingolipidome, sensitive, accurate, and comprehensive methods are needed. Here, we present a targeted reversed-phase liquid chromatography-high-resolution mass spectrometry-based workflow that significantly increases the accuracy of measured sphingolipids by resolving nearly isobaric and isobaric species; this is accomplished by a use of (i) an optimized extraction procedure, (ii) a segmented gradient, and (iii) parallel reaction monitoring of a sphingolipid specific fragmentation pattern. The workflow was benchmarked against an accepted sphingolipid model system, the RAW 264.7 cell line, and 61 sphingolipids were quantified over a dynamic range of 7 orders of magnitude, with detection limits in the low femtomole per milligram of protein level, making this workflow an extremely versatile tool for high-throughput sphingolipidomics.
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000131644 7001_ $$aWeintraub, Susan T$$b1
000131644 7001_ $$00000-0002-3771-2410$$aComan, Cristina$$b2
000131644 7001_ $$aPonnaiyan, Srigayatri$$b3
000131644 7001_ $$0P:(DE-He78)712670f67b229b25980c0e1c88ae8a09$$aSharma, Rakesh$$b4$$udkfz
000131644 7001_ $$0P:(DE-He78)a33ae52a1d80b847405db3ab83b9e90d$$aTews, Björn$$b5$$udkfz
000131644 7001_ $$00000-0001-6788-6641$$aWinter, Dominic$$b6
000131644 7001_ $$00000-0003-0232-3375$$aAhrends, Robert$$b7
000131644 773__ $$0PERI:(DE-600)1483443-1$$a10.1021/acs.analchem.7b03576$$gVol. 89, no. 22, p. 12480 - 12487$$n22$$p12480 - 12487$$tAnalytical chemistry$$v89$$x1520-6882$$y2017
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