Fabricating a Three-Dimensional Surface-Enhanced Raman Scattering Substrate Using Hydrogel-Loaded Freeze-Induced Silver Nanoparticle Aggregates for the Highly Sensitive Detection of Organic Pollutants in Seawater

Surface-enhanced Raman scattering (SERS) spectroscopy faces challenges in achieving both high sensitivity and reproducibility for the detection of real samples, particularly in high-salinity matrices. In this study, we developed a high-performance, salt-resistant three-dimensional (3D) SERS substrate by integrating physically induced colloidal silver nanoparticle aggregates (AgNAs) with an agarose hydrogel. AgNAs were prepared using a freeze-thaw-ultrasonication method to minimize interference in SERS signals while significantly enhancing the detection efficiency of trace pollutants. The incorporation of the agarose hydrogel not only improved the substrate's pollutant enrichment capability, but also effectively prevented the aggregation and sedimentation of AgNAs in salt solutions. The developed SERS substrate exhibited an ultralow detection limit of 10^-12 M for Nile Blue (NB), with a 100-fold increase in sensitivity compared to colloidal AgNAs and drop-cast AgNAs solid substrates. The analytical enhancement factor (AEF) for malachite green (MG) achieved a value of 1.4 × 10⁷. Furthermore, the substrate demonstrated excellent signal uniformity, with a relative standard deviation (RSD) of 6.74% within a 200 μm × 200 μm detection area and also show a satisfactory RSD of only 9.38% within a larger area of 1 mm × 1 mm. Notably, the 3D SERS substrate exhibited excellent stability under high-salinity conditions (0.5 M NaCl) and successfully detected a model pollutant (MG) in real seawater samples using the standard addition method. This study provides a novel strategy for highly sensitive SERS detection of trace pollutants in saline environments, offering promising applications in environmental monitoring and marine pollution analysis.