on 08-03-201605:48 AM - edited on 10-15-202105:14 AM by AnalyteGuru
Reiko Kiyonami 1, Claire Dauly 2, Ralf Tautenhahn1, David Peak1 and Ken Miller1
Metabolomics has become essential for understanding cell biology, physiology and medicine by providing a direct functional readout of cellular biochemical state. Recent advances in UHPLC separation, high resolution accurate mass spectrometers and metabolite database annotations have allowed the rapid and sensitive detection of a variety of metabolites from biological samples with minimal sample preparation. However, metabolite identification from LC-MS based data sets remains challenging due to the presence of many isomeric and isobaric metabolites In order to confidently identify metabolites high quality MS/MS or MSn data are required for many of the m/z features captured during a metabolomics profiling experiment.
The new Thermo ScientificTM Orbitrap Fusion TM Lumos TM Tribrid TM mass spectrometer offers high sensitivity and fast scan speed (up to 20 Hz), enabling comprehensive coverage of MS/MS spectra of detected m/z features in a LC-MS-MS run. It also enables multiple dissociation techniques, including HCD and CID, to be performed in parallel during a single LC-MS-MS run, providing comprehensive fragment information per m/z feature for confident metabolite identification. In this study, we demonstrate that high metabolite coverage and precise relative quantitation on two phenotypes of human urine samples can be achieved by combined three data dependent (dd) MS-MS analysis (dd HCD MS-MS with inclusion list, dd HCD MS-MS with exclusion list, dd alternated HCD-CID MS-MS with inclusion list) using a Orbitrap Fusion Lumous MS. The Thermo ScientificTM Compound Discoverer TM 2.0 (CD 2.0) is used for confident metabolite identification as well as differential and statistical analysis. Here we report that the metabolite identification coverage is increased by combining multiple MS-MS acquisitions while the precise analytical precision is simultaneously achieved by using fixed cycle time.
1. Thermo Fisher Scientific, San Jose, CA 95134
2. Thermo Fisher Scientific, Courtaboeuf, France