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A novel fluorescein-based probe <b>FLA-Boe</b> was developed for detecting H₂O₂. Modified by 2-Bromomethylphenylboronic acid pinacol ester, <b>FLA-Boe</b> is a Fluorescein derivative with eminent photostability and remarkable H₂O₂ sensitivity and selectivity. <b>FLA-Boe</b> was utilized to synthesize hydrogel sensors in the manner of guest–host interaction by taking advantage of its aforementioned features. The hydrogel sensor can be used to detect H₂O₂ effectively in both flowing and static water environments with satisfactory performance. It is expected that this application may open a new page to develop a neoteric fluorescent property analysis method aiming at H₂O₂ detection.