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Abstract
Probing the entirety of any species metabolome is an analytical grand challenge, especially at a cellular scale. Where spatial metabolomics, completed primarily by matrix-assisted laser desorption/ionization (MALDI), has limited molecular coverage for several reasons. To expand the scope of spatial metabolomics, we developed an on-tissue chemical derivatization (OTCD) workflow using 4-APEBA for confident identification of several dozen elusive phytocompounds, including several phytohormones, which have various roles within stress responses and cellular communication. Superiority of 4-APEBA is established in comparison to other derivatization agents with (1) broad specificity towards carbonyls, (2) low background, and (3) introduction of bromine isotopes, where the latter two facilitate confident bioinformatics. The outlined workflow trailblazes a path towards spatial hormonomics within plant samples, enhancing detection of carboxylates, aldehydes, ketones, and plausibly phenols.
Supplementary materials
Title
Supporting Information
Description
Materials and Methods:
Supporting Figure S1: MALDI-CID of derivatized standards of carboxylic acids and lignin decay products.
Supporting Figure S2: Ion images of in-situ derivatized exemplary natural product standards.
Supporting Figure S3: Additional ion images from derivatization performed within a solution of 50% MeOH.
Supporting Figure S4: NOR vs DHB. MALDI matrix effect on sensitivity and molecular coverage of 4-APEBA in-situ derivatization.
Supporting Figure S5: Derivative abscisate within METASPACE in poplar roots after 4-APEBA OTCD.
Supporting Figure S6: Non-derivatized vs derivatized molecules. Comparison of spatial patterns of exemplary metabolites with and without in-situ derivatization.
Supporting Figure S7: 1H-NMR spectrum of synthesized APEBA.
Supporting Figure S8: 1H-NMR spectrum of synthesized BTA.
Supporting Table S1: Characteristic neutral losses within Supporting Figure S1.
Supporting Table S2: Variable on tissue chemical derivatization conditions.
Supporting Table S3: Additional details within Supporting Figure S2.
Actions
Title
Supporting Workbook
Description
Complete list of METASPACE annotations from multiple databases from the imaging results.
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