The current study included collection of 90 serum samples to assess the relationship between the level of vitamin D3 and some biochemical variables in women with polycystic ovarian syndrome. The samples were collected from external laboratories in Salah al-Din Governorate for the period from 10/29/2021 to 12/25/2021. 60 samples were taken for women with PCOS, and 30 samples were taken from the control group, their ages ranged between (18-40) years for healthy women and patients. Vitamin D3 level and some hormones concentrations (luteinizing hormone, follicle-stimulating hormone, and testosterone) were estimated in the blood serum of the study groups. The results of the current study showed that the level of vitamin D3 decreased significantly (P≤0.05) in women with PCOS compared to healthy women as a control group, the results showed a significant increase (P≤0.05) in the level of studding hormones (luteinizing hormone, follicle-stimulating hormone, and testosterone) in women with PCOS compared to healthy women as a control group.