Utilizing Hydrogel-Isolated Dual-Carboxyl Surface-Enhanced Raman Scattering Probe for Accurate Online pH Monitoring

Surface-enhanced Raman scattering (SERS)-based pH sensors have been widely applied; However, the used 4-mercaptobenzoic acid (4-MBA) probes exhibit small pH-sensitive peak changes (carboxyl group) and strong susceptibility to interference, leading to inaccurate measurements. To address these limitations, we developed a SERS pH sensor using 2,5-dimercaptoterephthalic acid (2,5-DMTA) as the probe, which contains dual carboxyl groups. These carboxyl groups undergo reversible protonation-deprotonation, producing pronounced and reproducible spectral responses that enhance detection accuracy. The developed sensor enabled reliable detection across the acidic pH range of 0-7, showing good linearity (R² = 0.9786) and compensating for the weak acidic response of 4-MBA. Importantly, the 2,5-DMTA-based pH sensor demonstrated much better detection accuracy (detection relative standard deviation RSD less than 5%) than the 4-MBA-based SERS pH sensor (detection RSD ≈ 20%). To further improve measurement accuracy in complex matrices, the sensor was embedded in hot agarose to form an AuNP@hydrogel substrate, effectively suppressing interference from small molecules. Moreover, the developed sensor also shows satisfactory online pH monitoring features, including good reversibility (≥6 cycles), high stability of continuous measurements (30 min), and long-term storage stability (30 days). Integrated with a microfluidic 3D-printed flow cell, the system enabled rapid response (∼120 s) and online pH monitoring, and was successfully applied to continuous testing in lake water. Overall, this SERS platform provides a robust and accurate solution for SERS-based pH detection under acidic and complex environmental conditions.