Abstract
This study aims to develop an innovative electrochemical genosensor based on activated biochar (ABC) derived from the biomass of the seaweed Sargassum spp. The synthesis process begins with the pyrolysis of Sargassum spp. at 500 °C to obtain biochar (BC), which is chemically activated with nitric acid (HNO(3)). 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 m(2)/g (BC) to 159.2915 ± 2.2641 m(2)/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.