Covalent-Bonded Reduced Graphene Oxide–Fluorescein Complex as a Substrate for Extrinsic SERS Measurements

When graphene is used as SERS substrates, it contributes to the chemical mechanism (CM) of enhancement of Raman signal, owing to which the detection limit is very low (lower than mM content of probe molecules). The CM of enhancement depends largely on the interactions between the substrate and the probe molecules. Therefore, in this work, we have investigated the possibility of increasing the SERS activity of graphene by improving the interaction between the probe molecule and the graphene substrate by establishing exclusively strong covalent bonding between them. Fluorescein (Fl) was selected as a probe molecule because it is one of the most commonly used fluorophore in bioscience. As a graphene substrate, reduced graphene oxide (rGO) platelets were used. In addition, silver nanoparticles (AgNPs) were added onto the hybrids to further increase the enhancement by electromagnetic mechanism. Highly enhanced Raman signal of Fl onto neat rGO was achieved for micromolar concentration of the probe molecules. This was attributed to the covalent bonding between them, which introduced hole doping to rGO, decreasing the Fermi level of rGO and bringing it more closely to the LUMO of Fl. This induces aligning of their energy levels, resulting in higher contribution of the nonresonance effect to the charge transfer mechanism of enhancement, which, in this case, occurred intramolecularly. When AgNPs were added onto the rGO substrate, the expected enhancement performance was not observed. On the one hand, this was attributed to small size (∼20 nm) of AgNPs and lack of aggregates and, on the other, due to the unusually high contribution of CM determined.The financial support of the Basque Government (GV IT999- 16, IE16/44, and IE16/78), the Spanish Government (Ministerio de Economía y Competitividad project CTQ2016-80886-R) and the University of the Basque Country UPV/EHU (UFI11/56) is gratefully acknowledged. N.A. acknowledges the financial support from the EU Graphene Flagship project, Work Package 5, Biomedical Technologies (no. 696656)