Although there are various ways to make nanoparticles, the simple, risk-free, and affordable approaches are more significant and beneficial. Pulse laser ablation in liquid (PLAL) is a crucial technique that stands out for its affordability, safety, and simplicity. As a colloidal solution suspension, PLAL has generated Au NPs and Ag NPs. According to XRD studies, the cubic phase and small crystallite size of Au NPs is (18.6) nm, Ag NPs have been synthesized with the same energy, according to XRD studies, the cubic phase and small crystallite size of Ag NPs is (20.85) nm, to examine the morphology of particles, employ transmitted electronic microscopic (TEM), it was so clear that conjugated Levofloxacin with Au NPs making a random shape and the zones, AuNPs caping by Levofloxacin around the surface, so the average particle size as shown in the histogram size distribution varied to nearly 36nm. the conjugating of Levofloxacin on the surface of Ag NPs leads to making a random shape and the zones, so the average particle size as shown in the histogram size distribution varied to nearly 69 nm. Ultraviolet-Visible spectroscopy (UV-Vis) is used to display the absorption peaks, and, Fourier-transform infrared spectroscopy (FTIR) is used to investigate the different kinds of binding bonds, in turn, explained the difference after conjugation. One of the more important fields is biomedical use, so the antibacterial activity of Au NPs with Levofloxacin and Ag NPs with Levofloxacin appears to be enhanced by large inhibition zones for (Au- Levofloxacin Nano conjugate, Ag- Levofloxacin Nano conjugate) against E. coli, Staphylococcus aureus, Pseudomonas aeuginosa, and Proteus vulgaris, Additionally, it is more efficient than Levofloxacin, Au NPs, and Ag NPs took separately.