pH-Sensitive fluorescent sensor for Fe(III) and Cu(II) ions based on rhodamine B acylhydrazone: Sensing mechanism and bioimaging in living cells

Spirocyclic rhodamine derivatives have great potential to be used as fluorescent sensors. Rho- damine B hydrazide (RBH) and its derivatives have been employed to detect various analytes. The interactions of a sensor with an analyte might result in the protonation or hydrolysis of the sensor. Understanding these processes is useful for developing new sensors with improved characteristics. In this work, the performance of rhodamine B acylhydrazone (RBA) as a sensor for Fe3+ and Cu2+ ions is evaluated. In the presence of these ions, RBA undergoes protonation and the spirolactam ring opening. The ring opening renders the dye colored and fluorescent. RBA is then hydrolyzed to RBH leading to the decay of the absorbance in the visible range. The protonation and hydrolysis of RBA are acid-catalyzed, and metal ions contribute to these processes by lowering pH. Metal ions, unlike hydrogen ions, catalyze the transformation of RBH into rhodamine B and a phenanthrenone derivative. These products exhibit emission bands in the visible and near-infrared ranges, respectively. The obtained results can be applied to a variety of sensors based on rhodamines and Schiff bases. RBA can be employed for bioimaging. RBA quickly penetrates into cells, localizes in the organelles with acidic pH, probably in lysosomes, persists there for a long time, and gives bright fluorescence in the visible range. Cell incubation with Cu2+ ions produces fluorescence in the near-infrared range. RBA can be used as a multifunctional fluorescent biosensor to visualize cell compartments with acidic pH and detect Cu2+ ions in living cells.