A dual-functional rhodamine B and azo-salicylaldehyde derivative for the simultaneous detection of copper and hypochlorite: synthesis, biological applications and theoretical insights†

Abstract

A multifunctional rhodamine derivative containing azo-salicylaldehyde (BBS) was designed and synthesized as a colorimetric and fluorescence turn-on probe for the selective detection of copper cations (Cu²⁺) and hypochlorite anions (OCl⁻) in aqueous media. In the presence of Cu²⁺, the probe BBS exhibited turn-on absorption and fluorescence change at 554 nm and 585 nm, respectively. The binding mechanism of BBS with Cu²⁺ induces the opening of a spirolactam ring in the rhodamine moiety by the formation of a metal–ligand complex, achieving 10-fold enhancement in fluorescence and quantum yield, along with a binding constant of 1 × 10⁴ M⁻¹ and a detection limit of 2.61 μM. Addition of OCl⁻ enhanced the absorbance and fluorescence intensities at 520 nm and 575 nm, respectively. The probe BBS underwent hypochlorite-mediated oxidation, followed by hydrolysis, resulting in the formation of rhodamine B itself, which is detectable by the naked eye via the color and fluorescence enhancement by 11-fold with a high quantum yield and a detection limit of 1.96 μM. For practical applications, sensor BBS can be used to detect Cu²⁺ in water samples and on cotton swabs. For biological applications, the interaction of the BBS–Cu(II) complex with transport proteins such as bovine serum albumin (BSA) and ct-DNA was investigated through UV-vis and fluorescence titration experiments. Additionally, the structural optimization of BBS and the BBS–Cu(II) complex was demonstrated using DFT, and the interactions of the BBS–Cu(II) complex with BSA and ct-DNA were analysed through theoretical docking studies. Bioimaging studies were conducted by capturing fluorescence images of BBS with Cu²⁺ and OCl⁻ in a physiological medium containing living plant tissue using green gram seeds.