A complete workflow for improved untargeted metabolome annotation and identification using ultra high-resolution accurate mass and LC-MSn Orbitrap-based mass spectrometry

[Orbitrap_SciLib]

David A. Peake, Reiko Kiyonami, Ioanna Ntai, Amanda Souza and Ralf Tautenhahn

Purpose: Demonstrate confidence in metabolite annotation from ultra-high resolution LC-MS and data dependent MS2 data using a combined global semi-targeted (GST) metabolomics workflow. Methods: NIST SRM1950 human plasma was analyzed using data dependent LC-MS/MS analysis and the AcquireX data acquisition strategy. A list of highly-confident annotations was prepared and 58 reference compounds were targeted for confirmation of metabolite annotations. Results: Comparing the results from standard mixtures, SRM1950 plasma and spiked plasma extracts provided unequivocal evidence that 56 out of the 58 compounds were identified.


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