In this study, two metal oxide nanoparticles NiO and MnO2 were synthesized from green sources Mentha spicata (M. spicata) extract and Malus domestica (M. domestica) peel extract, respectively. The optical and physical properties of the synthesized nanoparticles were characterized using spectroscopic and microscopic techniques. Simple, precise, and new spectrophotometric probes were suggested for the determination of three cephalosporin antibiotics, including levofloxacin (LVX), cephalexin (CPX), and cefotaxime sodium (CTX) in their pure form and commercial products. The spectrophotometric detection of the selected drugs is based on the catalytic enhancement of NiO and MnO2 nanoparticles (NPs) due to their unique optical properties. Linear relationships with main correlation coefficients 0.999 were obtained at 0.1–20, 1.0–80, and 0.001–100 µg mL−1 for the three drugs in the presence of NiONPs, whereas 0.01–60, 0.1–160, and 0.01–80 µg mL−1 were obtained in the presence of MnO2NPs at absorption wavelengths 290, 262, and 235 nm for LVX, CPX and CTX, respectively. The analytical methods were validated and successfully used for determination of the instigated drugs in their bulk and commercial dosage forms.