The major an endocrine disorder in female is polycystic ovary syndrome (PCOS), characterized by anovulation, irregular menstruation, amenorrhea, hirsutism, and infertility. Recently, interest has increased in studying the effects of Vit. D in many diseases, including PCOS. The goal of this study was to estimate the link of Vit D and the risk of PCOS by measuring estrogen, progesterone, follicle-stimulating hormone (FSH), ovulation hormone, or called luteinizing hormone (LH), thyrotropin (TSH), total cholesterol, triacylglycerol TG, high-density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), and vitamin D in serum of 100 infertile with PCOS, 60 infertile with non-PCOS, and 60 control group. The results showed increase significantly in the concentration of estrogen, LH, TSH, cholesterol, triacyclglycerol, VLDL, and LDL compared to healthy women at P=0.01 (82.1±33.1 pg/ml), P=0.01 (5.71±3.34 mlU/ml), P=0.001 (2.48±1.23 µlU/ml), P=0.001 (186.3±39.4 mg/dL), P=0.001 (166.3±86.7 mg/dL), P=0.01 (33.4±17.2 mg/dL), and P=0.01 (112.6±34.5 mg/dL), respectively. Further, there was indeed decrease significantly in the concentration of progesterone, FSH, Vit D, and HDL compared to healthy women at P=0.01(1.295±0.82 ng/ml), P=0.001 (4.83±2.9 mlU/ml), P=0.001(11.94±5.82 mg/dL), and P=0.01 (40.83±8.80 mg/dL), respectively. The study showed that there is a strong correlation between vitamin D and the hormonal and biochemical variables that were measured. Therefore, we conclude that Vit. D is a new indicates the increase of infertility with PCOS.