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A novel quinazolinone based turn-on fluorescence probe for sensitive monitoring hypochlorite was prepared using the mild condensation reaction between 2-(2′-hydroxyphenyl)-4(3H)-quinazolinone derivative and 4-methylbenzenesulfonyl hydrazide. The probe exhibited specific selectivity to ClO− with obvious optical signal changes from weak fluorescence at 560 nm to a strong fluorescence emission at 520 nm and color changes from colorless to yellow, which could be noticed by the naked eye. The detection limit toward hypochlorite is as low as 11.4 nM. Moreover, the probe could sensitively response to ClO− in living cells with satisfying imaging effect and has been successfully applied to the determination of ClO− in practical water samples, which indicated that the probe has certain application potential for hypochlorite monitoring. Unlabelled Image • A quinazolinone based fluorescent probe for detection of ClO− was designed and synthesized. • Probe QBH showed a rapid and remarkable fluorescence "turn-on" response to ClO−. • The QBH could detect ClO− quantitatively with a detection limit as low as 11.4 nM. • The sensing mechanism is analyzed through the NMR and MS spectra. • The probe was successfully applied to the determination of ClO− in practical water samples. [ABSTRACT FROM AUTHOR]