Authors
Frederico G Pinto, Alexander D Giddey, Nesrin Mohamed, Rauda S B Almarri, Munazza Murtaza, Nasna Nassir, Omer S Alkhnbashi, Mohammed J Uddin, Nelson C Soares
Published in
Expert review of proteomics. Jul 21, 2025. Epub Jul 21, 2025.
Abstract
Many of the advanced MS methods applied in proteomics such as nanoflow LC-MS with data-independent acquisition have yet to be verified and/or optimized on metabolomics applications.
This study evaluates the feasibility of repurposing a proteomics-optimized nanoLC-MS platform for untargeted metabolomics. Using NIST SRM 1950 reference human plasma, we compared the performance of polarity switching and separate polarity modes under DIA conditions, focusing on metabolite coverage, annotation, and response linearity.
We observed, in the separate polarity and switching polarity runs 669 and 353 features in (+) mode and 558 and 446 features in (-) mode, respectively. A total of 233 metabolites were annotated using the (±) separate polarities and 179 using the (±) switching polarity based on MassBank of North America (MoNA) public MS library and filtered with the Human Metabolome Database (HMDB). Both switching and separate polarity methods performed well regarding response linearities which were investigated by spiking some amino acid compounds into plasma matrix.
The polarity switching DIA approach for metabolomics reduced sample consumption and analysis time, but led to fewer detected features and annotations compared to separate polarity runs. These findings support the use of unified nanoLC-MS platforms for integrated multi-omics analysis.
PMID:
40685892
Bibliographic data and abstract were imported from PubMed on 21 Jul 2025.
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