Cyanine based ratio fluorescent probe and its application in hypochlorite detection

Hypochlorite (ClO−), a weakly acidic reactive oxygen species, plays a crucial role in antibacterial and anti-inflammatory defense mechanisms. However, elevated levels of ClO− or disruptions in endogenous sites can lead to tissue damage and various diseases including cardiovascular disease, neuronal degeneration, and arthritis. To address this, the development of a specific fluorescent probe with a built-in self-calibration ratio mode for the analysis and biological imaging of ClO− is essential. In this study, a cyanine-based fluorescent probe (Cy-H) was designed for ratiometric fluorescent detection of ClO−, utilizing its aggregation behavior as a novel approach in this field. Upon exposure to ClO−, the phenolic hydroxyl group in probe Cy-H was oxidized into benzoquinone, leading to the formation of cyanine products that displayed a strong tendency to aggregate. As a result, the maximum emission peak of the probe shifted from 700 nm to 485 nm. Notably, a linear relationship was observed between the peak intensity ratio (I485/I700) and the concentration of hypochlorite, with a limit of detection (LOD) of 0.49 μM. Furthermore, this probe was successfully employed for imaging analysis of hypochlorite in living cells and zebrafish.