Expanded Coverage of Phytocompounds by Mass Spectrometry Imaging Using On-Tissue Chemical Derivatization by 4-APEBAClick to copy article linkArticle link copied!
- Kevin J. ZemaitisKevin J. ZemaitisEarth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington 99354, United StatesMore by Kevin J. Zemaitis
- Vivian S. LinVivian S. LinEarth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington 99354, United StatesMore by Vivian S. Lin
- Amir H. AhkamiAmir H. AhkamiEarth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington 99354, United StatesMore by Amir H. Ahkami
- Tanya E. WinklerTanya E. WinklerEarth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington 99354, United StatesMore by Tanya E. Winkler
- Christopher R. AndertonChristopher R. AndertonEarth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington 99354, United StatesMore by Christopher R. Anderton
- Dušan Veličković*Dušan Veličković*Email: [email protected]Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, Washington 99354, United StatesMore by Dušan Veličković
Abstract
Probing the entirety of any species metabolome is an analytical grand challenge, especially on a cellular scale. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) is a common spatial metabolomics assay, but this technique has limited molecular coverage for several reasons. To expand the application space of spatial metabolomics, we developed an on-tissue chemical derivatization (OTCD) workflow using 4-APEBA for the confident identification of several dozen elusive phytocompounds. Overall, this new OTCD method enabled the annotation of roughly 280 metabolites, with only a 10% overlap in metabolic coverage when compared to analog negative ion mode MALDI-MSI on serial sections. We demonstrate that 4-APEBA outperforms other derivatization agents by providing: (1) broad specificity toward carbonyls, (2) low background, and (3) introduction of bromine isotopes. Notably, the latter two attributes also facilitate more confidence in our bioinformatics for data processing. The workflow detailed here trailblazes a path toward spatial hormonomics within plant samples, enhancing the detection of carboxylates, aldehydes, and plausibly other carbonyls. As such, several phytohormones, which have various roles within stress responses and cellular communication, can now be spatially profiled, as demonstrated in poplar root and soybean root nodule.
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- Kevin J Zemaitis, Christopher R Anderton, Dusan Velickovic. A selective process for application of EDC and 4-APEBA for cabonyl containing metabolites by MALDI-MSI v1. 2024https://doi.org/10.17504/protocols.io.ewov19d9ylr2/v1
- Kevin J Zemaitis, Christopher R Anderton, Dusan Velickovic. A selective process for application of EDC and 4-APEBA for carbonyl containing metabolites by MALDI-MSI v2. 2024https://doi.org/10.17504/protocols.io.ewov19d9ylr2/v2
- Zhi Sun, Fangfang Wang, Yuwei Liu, Bowen Deng, Ruobing Ren, Yifei Wang, Di Chen, Lihua Zuo. Recent strategies for improving MALDI mass spectrometry imaging performance towards low molecular weight compounds. TrAC Trends in Analytical Chemistry 2024, 175 , 117727. https://doi.org/10.1016/j.trac.2024.117727
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- Dušan Veličković, Kevin J. Zemaitis, Arunima Bhattacharjee, Christopher R. Anderton, . Mass spectrometry imaging of natural carbonyl products directly from agar-based microbial interactions using 4-APEBA derivatization. mSystems 2024, 9
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