Lipid Profiling of Secreted Lung Lipids by MSMSALL Shotgun Lipidomics
November 18, 2021
Recent advances in the analytical field, mainly mass spectrometry, has fueled lipidomics research. Data independent acquisition (DIA) based MS analysis is now considered as a go-to approach for performing lipidomics analysis. Among the DIA-based approaches, the sequential precursor ion fragmentation, MSMSALL, has shown prominence in performing both quantitative and qualitative lipid analysis. In this approach, the quadrupole Q1 transmits ions (wide mass range, 1 amu) consisting of multiple analytes through the collision cell. These transmitted ions are subjected to fragmentation and are analyzed in a TOF-MS analyzer at higher resolution. The MS/MS spectrum generated comprises a composite of all analytes within the Q1 m/z window.
Data analysis of this complex spectrum has been challenging. To overcome this complexity, SimLipid® software offers a comprehensive workflow using automated large volume loop injection (FIA) coupled with a SCIEX Triple TOF mass spectrometer.
Samples from mouse bronchoalveolar lavage that were lipopolysaccharide treated and control mice were extracted and reconstituted. Using the automated loop injection, these samples were subjected to the SCIEX Triple TOF instrument for MSMSALL acquisition. The MSMSALL acquisition was performed with 1 Da selection window under positive ion mode. The acquired data was submitted to SimLipid for automatic identification of the lipids using product ion data. MS/MS database search was performed using 0.5 Da for precursors and 0.01 Da for product ions as error tolerance. A total of 452 unique lipids were identified from the search, and a custom database was created using these lipids. For quantitation, the TOF-MS data from two biological groups were imported into SimLipid by averaging the TOF-MS scans with 5 ppm tolerance. The exact mass search was performed on these two groups using the custom database. The results were exported for further statistical analysis using other third party software tools.
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