Conventional extraction-based phytochemical analysis can identify many plant constituents but removes the anatomical context needed to determine where specialized metabolites accumulate, co-occur, or change within medicinal and bioactive plants. Mass-spectrometry imaging (MSI) addresses this spatial gap, although ion images differ substantially in chemical-identification confidence, quantitative meaning, and biological interpretability. This scoping review mapped peer-reviewed Q1 literature published from 2017 through 2024 on MSI-based localization of specialized metabolites in medicinal or bioactive plants, together with a limited set of transferable analytical and reporting studies required to interpret those applications. A DOI-unique candidate set of 52 records was screened using predefined eligibility and evidence-fit criteria, yielding 37 included articles. Study characteristics were charted across plant material, spatial scale, metabolite class, imaging modality, orthogonal validation, biological question, uncertainty, and transferability. The evidence shows a methodologically diverse field dominated by problem-specific analytical choices rather than a single preferred platform. MALDI-, DESI-, SALDI-, ambient-, and surface-aware approaches addressed different constraints involving tissue geometry, low-mass background, volatility, ionization, isomerism, and quantitation. Applications ranged from tissue-specific biosynthesis and developmental localization to cultivation, processing, material differentiation, and quality characterization. Stronger biological claims consistently depended on evidence beyond spatial co-localization. MSI can make phytochemistry anatomically explicit, but localization, molecular identity, abundance, and biological explanation should be treated as distinct evidential claims. The literature supports expanding spatial phytochemistry in medicinal plants while prioritizing claim-matched validation, reproducible sampling, and explicit interpretation boundaries.
