Objectives: Sulphur is among the most common minerals in the body mainly in proteins, but, also, commonly found in tissues joints and cells .At the same time there is a rapidly expanding literature on the effects, of sulphur in regulating biologicaly systems. Methods: Preparation of sulfur nanoparticles (S-NPs) was preparation and fabrication of nanosulfar were performed. Firstly, the evaluation of their composition and purity by fourier transform infrared spectrometer (FT-IR) analysis. Secondly, The shape and size of S-NPs were investigated by scanning electron microscopy (SEM) techniques in all S-NPs samples were size 50nm, 10nm and 5 nm , Then, Energy-dispersive X-ray spectroscopy (EDS) for the evaluation of their composition and purity, after that the X-Ray diffraction, and the determination of S-NPs antitumor activity against (MCF-7, HepG2, HCT116, and PC3) cell lines and were minimum inhibitory concentrations found to be 10.7 ng/ml, 3.7 ng/ml ,10.6ng/ml, and 3.34 ng/ml; respectively. Finaly, determination biochemical assay of S-NPs by using female mice bearing EAC and of the assessment of liver functions, kidney functions, heart functions and hematology markers. Results: This study revealed that S-NPs showed high strong activities in concentration (5 mg/kg) as it improvement of liver functions production compared to the positive control group, also showed the effect of S-NPs of the kidney functions , heart functions and hematology parameters in all studied groups and a significant increase in activities compared with positive control group.