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In situ droplet-based on-tissue chemical derivatization for lipid isomer characterization using LESA

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Abstract

In this work, we present an in situ droplet-based derivatization method for fast tissue lipid profiling at multiple isomer levels. On-tissue derivatization for isomer characterization was achieved in a droplet delivered by the TriVersa NanoMate LESA pipette. The derivatized lipids were then extracted and analyzed by the automated chip-based liquid extraction surface analysis (LESA) mass spectrometry (MS) followed by tandem MS to produce diagnostic fragment ions to reveal the lipid isomer structures. Three reactions, i.e., mCPBA epoxidation, photocycloaddition catalyzed by the photocatalyst Ir[dF(CF3)ppy]2(dtbbpy)PF6, and Mn(II) lipid adduction, were applied using the droplet-based derivatization to provide lipid characterization at carbon–carbon double-bond positional isomer and sn-positional isomer levels. Relative quantitation of both types of lipid isomers was also achieved based on diagnostic ion intensities. This method provides the flexibility of performing multiple derivatizations at different spots in the same functional region of an organ for orthogonal lipid isomer analysis using a single tissue slide. Lipid isomers were profiled in the cortex, cerebellum, thalamus, hippocampus, and midbrain of the mouse brain and 24 double-bond positional isomers and 16 sn-positional isomers showed various distributions in those regions. This droplet-based derivatization of tissue lipids allows fast profiling of multi-level isomer identification and quantitation and has great potential in tissue lipid studies requiring rapid sample-to-result turnovers.

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Funding

The authors gratefully acknowledge the NIH NIGMS Maximizing Investigators’ Research Award R35GM143047, NIH P41GM128577, NIH/NIA 1R01AG064869, and the Welch grant A-2089 for financial supports.

All experimental procedures were approved by the Institutional Animal Care and Use Committee at Texas A&M University (IACUC 2022–0180), and all methods were performed in accordance with the relevant guideline and regulations.

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Authors and Affiliations

  1. Department of Chemistry, Texas A&M University, 580 Ross St, College Station, TX, 77843, USA

    Dallas P. Freitas, Xi Chen, Erin A. Hirtzel, Madison E. Edwards, Joohan Kim, Klaudia I. Kocurek, David Russell & Xin Yan

  2. Department of Nutrition, Texas A&M University, Carter-Mattil Hall, 373 Olven Blvd, College Station, TX, 77843, USA

    Hongying Wang & Yuxiang Sun

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  1. Dallas P. Freitas
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Published in the topical collection Young Investigators in (Bio-)Analytical Chemistry 2023 with guest editors Zhi-Yuan Gu, Beatriz Jurado-Sánchez, Thomas H. Linz, Leandro Wang Hantao, Nongnoot Wongkaew, and Peng Wu.

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Freitas, D.P., Chen, X., Hirtzel, E.A. et al. In situ droplet-based on-tissue chemical derivatization for lipid isomer characterization using LESA. Anal Bioanal Chem 415, 4197–4208 (2023). https://doi.org/10.1007/s00216-023-04653-3

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