What does this strategy enable?
Mass spectrometry imaging–guided molecular networking connects where a metabolite occurs with how its fragmentation spectrum relates to other molecules. Spatially resolved MSI features are used to identify chemically informative regions or ions, while LC-MS/MS molecular networking supplies fragmentation-based family context and annotation. The combination can focus structural analysis on spatially localized natural products rather than treating imaging and molecular networking as independent readouts.
Transferable workflow
Workflow
- 1
Acquire spatial metabolite maps
Use MALDI or another suitable imaging-MS method to map metabolite ions across the tissue or material of interest.
- 2
Prioritize spatially informative features
Select ions or chemical regions whose localization suggests biological or anatomical relevance.
- 3
Acquire complementary LC-MS/MS data
Generate fragmentation-rich LC-MS/MS data from matched or relevant material so spatially prioritized chemistry can be interpreted structurally.
- 4
Build molecular networks
Organize related MS/MS spectra into molecular families and connect the spatially prioritized features to family-level structural context.
- 5
Validate spatial and structural assignments
Confirm prioritized annotations and localization with orthogonal standards, fragmentation interpretation, or additional imaging experiments where possible.
Connected resources
Tools used in this strategy
Strategy profile
Related objectives
Natural-product discovery, Structure elucidation, Molecular-family discovery
Components
Sample preparation, Data acquisition, Molecular networking, Annotation or identification, Biological validation
Sample types
Plant tissue, Tissue or biopsy
Biological context
Plant, Broadly applicable
Ion mobility
Ion mobility not required
Scope and considerations
Designed for
- Natural-product or specialized-metabolite discovery where spatial localization can guide which chemistry deserves structural follow-up.
- Multimodal studies combining imaging MS with fragmentation-rich LC-MS/MS.
Important requirements
- Spatially resolved MSI data and complementary MS/MS data that can be related to the same chemistry.
Considerations
- Matching an MSI ion to an LC-MS/MS feature requires careful mass, adduct, and sample-context reasoning.
- Spatial colocalization and spectral family membership do not substitute for complete structural confirmation.
Reusable resources
Publication
Kuo TH, Huang HC, Hsu CC. Mass spectrometry imaging guided molecular networking to expedite discovery and structural analysis of agarwood natural products. Analytica Chimica Acta 1080, 95–103 (2019).