Abstract: Novel nanoparticles were prepared by using Schiff’s reaction, to produce the nanoparticles Schiff bases (II and III) have been prepared by the composite reaction of graphene oxide and hydrazine hydrate with Isatin and Guanine respectively. On Other hand, Nanoparticle graphene oxide (I) was prepared by using Hummer's method that includes addition of graphite to a mixture of sodium nitrate and concentrated sulfuric acid in the presence of oxidizing agents like potassium permanganate and hydrogen peroxide. The biological activity of the synthesized nanocomposite was studied against various positive and negative gram bacteria by the use of Agar well diffusion method on Mueller - Hinton, the results have shown that the Nano Schiff base II has an interesting inhibition activity especially against the Staphylococcus aureus bacteria. On the other hand, Nano Schiff base III has shown moderate activity against Bacillus thyuringiensis at high concentration of 100µL and it exhibits low - zero inhibition activity against other tested bacteria.