Evaluation of Activated Biochar Derived from Sargassum spp. as a Sustainable Substrate for the Development of Electrochemical DNA Biosensing

This study aims to develop an innovative electrochemical genosensor based on activated biochar (ABC) derived from the biomass of the seaweed <i>Sargassum spp</i>. The synthesis process begins with the pyrolysis of <i>Sargassum spp</i>. at 500 °C to obtain biochar (BC), which is chemically activated with nitric acid (HNO₃). The physicochemical properties of the resulting material, such as morphology and surface area, were characterized using techniques including scanning electron microscopy (SEM), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and the Brunauer-Emmett-Teller (BET) method for surface area. BET results showed an increase in surface area from 22.9367 ± 0.0879 m²/g (BC) to 159.2915 ± 2.2641 m²/g (ABC). For the development of the genosensor, a hydrolyzed collagen gel matrix enriched with ABC is created. This nanostructured, biocompatible mixture is used to immobilize a DNA probe on a graphite electrode, employing the large surface area of ABC and the formation of a functional HC-based coating. The system's viability was evaluated by cyclic voltammetry (CV), which showed changes in the maximum anodic peak current (Ipa) during fabrication: 27.78 ± 1.87 μA for the bare electrode, 35.25 ± 1.24 μA for ABC 30%, and 39.25 ± 1.84 μA for HC + ABC 30%. After ssDNA immobilization and hybridization to dsDNA, Ipa decreased to 28.81 ± 1.565 μA and 23.10 ± 1.25 μA, respectively. Finally, hematoxylin (Hx) was used as an intercalating indicator from hybridization, reducing the maximum anodic peak current to 15.51 ± 1.13 μA, consistent with additional interfacial limitations associated with dsDNA formation. Overall, the developed system demonstrates a sustainable, promising platform for molecular diagnostics in electrochemical DNA biosensor development.