Overexposure to hypochlorous acid (HOCl) can lead to cellular oxidative stress. In response to the stress, the body activates related factor that upregulates a potent antioxidant enzyme, NAD(P)H:quinone oxidoreductase 1 (NQO1). A method that can simultaneously detect HOCl and NQO1 can help to understand their interactions during homeostasis. However, the methods are still limited due to the complexity of the biological environment. In this work, we present for the first time a dual-ratiometric fluorescent probe (NH) that can distinguish between HOCl and NQO1. Probe NH had ratiometric fluorescence responses by changing from red (λ_max^em = 643 nm) to green (λ_max^em = 517 nm) in the presence of HOCl and from red to mixed blue (λ_max^em = 483 nm)/red in the presence of NQO1 under single-wavelength excitation. This probe had high selectivity and sensitivity towards HOCl and NQO1 in vitro. Moreover, probe NH could specifically image exogenous or endogenous HOCl and NQO1 in living cells through different emission channels. This probe provides a practical tool for studying the correlation of HOCl and NQO1 in biological systems.

过度暴露于次氯酸 (HOCl) 会导致细胞氧化应激。为了应对压力，身体会激活相关因子，从而上调一种有效的抗氧化酶 NAD(P)H：醌氧化还原酶 1 (NQO1)。一种可以同时检测 HOCl 和 NQO1 的方法可以帮助了解它们在稳态过程中的相互作用。然而，由于生物环境的复杂性，这些方法仍然受到限制。在这项工作中，我们首次提出了一种可以区分 HOCl 和 NQO1的双比例荧光探针 ( NH )。探针NH通过从红色 (λ _max ^em  = 643 nm) 变为绿色 (λ _max ^{em ) 具有比率荧光响应} = 517 nm）在 HOCl 存在 下和在单波长激发下在 NQO1 存在下从红色到混合蓝色（λ _max ^{em = 483 nm）/红色。}该探针在体外对HOCl和NQO1具有高选择性和敏感性。此外，探针NH可以通过不同的发射通道特异性地成像活细胞中的外源性或内源性HOCl和NQO1。该探针为研究 HOCl 和 NQO1 在生物系统中的相关性提供了实用工具。