A Novel Carbon Dot-Bromothymol Blue System for Ratiometric Colorimetric-Fluorometric Sensing of Glutathione in Urine: A Smartphone-Compatible Approach.

Abstract

This study presents a novel dual-modal approach for glutathione (GSH) detection using blue and yellow dual-emission carbon dots (BY-CDs) and bromothymol blue (BTB) at pH 8.0. The method employs both colorimetric and fluorometric detection modes, offering a new perspective on GSH quantification. BTB's blue coloration induces selective fluorescence quenching of the CDs' 610 nm emission peak, with minimal effect on the 445 nm peak. Upon GSH addition, the quinonoid structure (blue color) of BTB transforms to its benzenoid form (yellow color). This transformation triggers fluorescence restoration at 610 nm and significant quenching at 445 nm, enabling ratiometric fluorescence analysis (F₆₁₀/F₄₄₅). Concurrently, colorimetric detection is also ratiometric, based on measuring the ratio between the emerging yellow color peak (435 nm) and the decreasing blue color peak (618 nm) (A₄₃₅/A₆₁₈). The state-of-the-art aspect of this detection method lies in the strategic choice of dual-emission CDs and a dye with distinct absorption spectra that closely match the emission spectra of the CDs. This unique combination allows for dual detection with opposite responses in the two detection modes, enhancing selectivity and reliability. The probe was thoroughly characterized, and its detection mechanism was elucidated using various spectroscopic techniques. The method shows excellent linearity, a broad detection range, and low detection limits for both fluorometry (0.02 - 10.0 μM, 5.88 nM) and colorimetry (1.0 - 35.0 μM, 301.25 nM). Additionally, a smartphone-based platform was developed for colorimetric GSH determination, enhancing the method's potential for on-site analysis. The assay's practicality was validated through successful application to human urine samples, yielding excellent recovery values (97.33% to 99.13%).