A Novel Ratiometric Photoelectrochemical Biosensor Based on Front and Back Illumination for Sensitive and Accurate Glutathione Sensing.

The ratiometric detection method has a strong attraction for photoelectrochemical bioanalysis due to its high reliability and real-time calibration. However, its implementation typically depends on the spatial resolution of equipment and the pairing of wavelength/potential with photoactive materials. In this paper, a novel ratiometric photoelectrochemical biosensor based on front and back illumination was prepared for the detection of glutathione (GSH). Unlike traditional ratio methods, this ratiometric biosensor does not require voltage and wavelength modulation, thereby avoiding potential crosstalk caused by voltage and wavelength modulation. Additionally, the formation of a heterojunction between mTiO and AgS is conducive to enhancing light absorption and promoting charge separation, thereby boosting the photocurrent signal. Apart from forming a heterojunction with TiO, AgS also shows a specific affinity towards GSH, thus enhancing the selectivity of the mTiO/AgS ratiometric photoelectrochemical biosensor. The results demonstrate that the ratiometric photoelectrochemical biosensor exhibits a good detection range and a low detection limit for GSH, while also possessing significant interference elimination capability. The GSH detection range is 0.01-10 mmol L with a detection limit of 6.39 × 10 mmol·L. The relative standard deviation of 20 repeated detections is 0.664%. Impressively, the proposed novel ratiometric PEC biosensor demonstrates enviable universality, providing new insights for the design and construction of PEC ratiometric sensing platforms.