Journal
JOURNAL OF PROTEOME RESEARCH
Volume 15, Issue 1, Pages 291-301Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.5b00841
Keywords
lipidomics; sphingolipids; liquid chromatography; software; high throughput; selected reaction monitoring
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Funding
- Ministry for Innovation, Science, and Research of the Federal State of North Rhine-Westphalia
- Federal Ministry of Education and Research in Germany
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In response to the urgent need for analysis software that is capable of handling data from targeted high-throughput lipidomics experiments, we here present a systematic workflow for the straightforward method design and analysis of selected reaction monitoring data in lipidomics based on lipid building blocks. Skyline is a powerful software primarily designed for proteomics applications where it is widely used. We adapted this tool to a Plug and Play system for lipid research. This extension offers the unique capability to assemble targeted mass spectrometry methods for complex lipids easily by making use of building blocks. With simple yet tailored modifications, targeted methods to analyze main lipid classes such as glycerophospholipids, sphingolipids, glycerolipids, cholesterylesters, and cholesterol can be quickly introduced into Skyline for easy application by end users without distinct bioinformatics skills. To illustrate the benefits of our novel strategy, we used Skyline to quantify sphingolipids in mesenchymal stem cells. We demonstrate a simple method building procedure for sphingolipids screening, collision energy optimization, and absolute quantification of sphingolipids. In total, 72 sphingolipids were identified and absolutely quantified at the fatty acid scan species level by utilizing Skyline for data interpretation and visualization.
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