A water-soluble fluorine magnetic resonance spectroscopy host-guest probe, P(HPA-co-AdamCFA), was successfully constructed from the facile synthesis of a bifunctional monomer via a quantitative Passerini reaction. Supramolecular complexation with (2-hydroxypropyl)-β-cyclodextrin promoted a change in the chemical environment, leading to modulation of both the relaxation properties as well as chemical shift of the fluorine moieties. This change was used to probe the supramolecular interaction by F MRI spectroscopy and give insight into fluorine probe formulation. This work provides a fundamental basis for an F MR imaging tracer capable of assessing host-guest inclusion and a potential model to follow the fate of a drug delivery system in vivo.