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Fenton chemistry has aroused widespread concern due to its application in the green oxidation and mineralization of organic wastes. Inorganic pyrophosphatase (PPase) catalyzes the hydrolysis of pyrophosphate ions (PPi) and provides a thermodynamic driving force for many biosynthetic reactions. Fluoride (F - ) is widely applied to fight against tooth decay and reduce cavities. The electrochemical determination of PPase activity and F - was realized based on Fenton chemistry in this work. Glassy carbon electrode modified with poly (azure A) and acetylene black (GCE/PAA-AB) was fabricated. Hydroxyl radicals (∙OH) that were generated from a Cu 2+ -catalyzed Fenton-type reaction could oxidize PAA in the near-neutral medium, leading to a great increase of the cathodic peak current (I pc ). A coordination reaction between PPi and Cu 2+ exerted a negative effect on Fenton reaction and hindered the I pc enhancement. Cu 2+ -PPi complex was decomposed due to the hydrolysis of PPi induced by PPase, which caused the reappearance of the notably increased current response. F - could effectively inhibit PPase activity. As a result, the stable Cu 2+ -PPi complex remained and the high I pc suffered from the decline again. The I pc difference was used for the highly sensitive determination of PPase activity in the content range of 0.001-20 mU mL -1 with a detection of limit (LOD) at 0.6 μU mL -1 and that of F - in the concentration range of 0.01-100 μM with a LOD at 7 nM. The proposed PPase and F - sensor displayed a good selectivity, stability and reproducibility, and a high accuracy., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest to this work., (Copyright © 2024 Elsevier B.V. All rights reserved.)