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Rellin, K.F. ; Abel, L.* ; Barsch, A.* ; Witting, M.

Optimizing nanoLC-TIMS-TOF and lipidomics sample preparation for low volume of plasma.

Sci. Rep. 16:30707 (2026)
Verlagsversion Forschungsdaten DOI
Open Access Gold
Creative Commons Lizenzvertrag
Lipidomics provides critical insights into disease mechanisms, biomarker discovery, and precision medicine, but conventional LC-MS workflows often require sample volumes that limit their application in minimally invasive studies. Here, we evaluated a nanoflow liquid chromatography workflow coupled with trapped ion mobility spectrometry and time-of-flight mass spectrometry (nanoLC-TIMS-TOF) for positive ion lipid profiling from as little as 1 µL of plasma. Two commonly used single-phase extraction protocols, MeOH: MTBE and IPA: H₂O, were scaled to microsample volumes using NIST® SRM® 1950 plasma. The effects of plasma input volume and mobile-phase ammonium formate concentration (2, 5, and 10 mM) on feature detection and chromatographic performance were evaluated. Descriptive comparisons revealed extraction-, volume-, and buffer-associated differences in the detected feature profiles, while putative lipid annotations were evaluated using MS/MS and ion-mobility information, where available. The optimized workflow enabled broad positive ion lipid class profiling and detection of low intensity features from only 1 µL of starting material. The 1 µL extracts retained the dominant annotated lipid classes observed at higher input volumes, although profiling depth and feature distributions remained dependent on plasma input volume. Among the tested mobile phase conditions, 5 mM ammonium formate produced the highest detected feature count while limiting pressure drift during extended acquisition. These results support the feasibility and practical scalability of positive ion nanoLC-TIMS-MS lipid profiling for sample-limited applications under the evaluated conditions.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Ion Mobility ; Lipidomics ; Nanolc ; Timstof ; Ultra-low Volume Sampling; Mass-spectrometry; Extraction; Stability; Lc
ISSN (print) / ISBN 2045-2322
e-ISSN 2045-2322
Zeitschrift Scientific Reports
Quellenangaben Band: 16, Heft: 1, Seiten: , Artikelnummer: 30707 Supplement: ,
Verlag Springer
Verlagsort London
Begutachtungsstatus Peer reviewed
Institut(e) Core Facility Metabolomics and Proteomics (CF-MPC)
Förderungen Helmholtz Zentrum Mnchen - Deutsches Forschungszentrum fr Gesundheit und Umwelt (GmbH) (4209)