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Magnetic resonance with hyperpolarized contrast agents is one of the most powerful and noninvasive imaging platforms capable for investigating in vivo metabolism. While most of the utilized hyperpolarized agents are based on 13 C nuclei, a milestone advance in this area is the emergence of 15 N hyperpolarized contrast agents. Currently, the reported 15 N hyperpolarized agents mainly utilize the dissolution dynamic nuclear polarization (d-DNP) protocol. The parahydrogen enhanced 15 N probes have proven to be elusive and have been tested almost exclusively in organic solvents. Herein, we designed a reaction based reactive oxygen sensor 15 N-boronobenzyl-2-styrylpyridinium ( 15 N-BBSP) which can be hyperpolarized with para-hydrogen. Reactive oxygen species plays a vital role as one of the essential intracellular signalling molecules. Disturbance of the H 2 O 2 level usually represents a hallmark of pathophysiological conditions. This H 2 O 2 probe exhibited rapid responsiveness toward H 2 O 2 and offered spectrally resolvable chemical shifts. We also provide strategies to bring the newly developed probe from the organic reaction solution into a biocompatible injection buffer and demonstrate the feasibility of in vivo 15 N signal detection. The present work manifests its great potential not only for reaction based reactive sensing probes but also promises to serve as a platform to develop other contrast agents., (© 2024 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.)