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In this work, the development of nanosystems by Pulsed Laser Ablation in Liquid (PLAIL) is of considerable importance to expand their biomedical applications, such as drug delivery. In the present study, we focus on the conditions of the preparation  Nd: YAG laser wavelength 266 nm and two different laser fluency (10,28) J/cm² to control the concentration and size stability of superparamagnetic iron oxide nanoparticles (SPION) prepared by PLAIL. The characteristics of SPION are investigated by energy-dispersive X-ray spectroscopy (EDX) spectra which showed strong peaks of Fe and O. Magnetic characteristics of iron oxide nanoparticles indicated superparamagnetic properties of SPION and suitable physical stability. Optical and chemical properties of SPION were investigated using UV-visible spectra (UV) and infrared Fourier transformed spectroscopy (FTIR). .  Scanning electron microscopy (SEM) was used to obtain surface morphological studies of SPION.. Results showed that SPION is the only cubic shape, the peak absorption shifted toward short wavelengths with optimum concentration to the SPION in double deionized water (DDW) and in Acetone (0.75,0.33) mg/ml respectively, at high laser fluence 28 J/cm², and this enhancement of value is due to particle size and color resultant in a solution. Eventually,, this product has the optimal SPION specialty ratio of SPION in the DDW solution at 53.89%, and the size is very suitable for drug delivery applications.