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Plasma-activated water (PAW) is considered to be an effective anticancer agent due to the diverse aqueous reactive oxygen and nitrogen species (RONS: ROS and RNS), but the drawback of low dose and short duration of RONS in acidified PAW limits their clinical application. Herein, this Letter presents an innovative therapeutic avenue for cancer treatment with highly-effective alkaline PAW prepared by air surface plasma. This anticancer alkaline formulation is comprised of a rich mixture of highly chemical RONS and exhibited a prolonged half-life compared to acidified PAW. The H2O2, NO2−, and ONOO−/O2− concentrations in the alkaline PAW can reach up to 18-, 16-, and 14-fold higher than that in acidic PAW, and the half-life of these species was extended over 8-, 10-, and 26-fold, respectively. The synergistic potent redox action between these RONS with alkaline pH was shown to be more potent than acidic PAW for cancer cell inhibition in vitro. Furthermore, the alkaline PAW injection treatment also significantly inhibited tumor growth in tumor-bearing mice. The possible reasons are that the alkaline PAW would disturb the acid extracellular milieu leading to the inhibition of tumor growth and progression; moreover, the efficient and durable RONS with alkaline pH could induce significant cell apoptosis by altering cell biomolecules and participating apoptosis-related signaling pathways. These findings offer promising applications for developing a strategy with real potential for tumor treatment in clinical applications. [ABSTRACT FROM AUTHOR]