The present study evaluates the effect of Vitamin D on infertility in females by measuring sex-related hormones (testosterone(TESTO), estradiol(E2), luteinizing hormone(LH,) and follicle-stimulating hormone(FSH)) and some biochemical markers (prolactin (PRL) and lipid profile). The study included 56femalese divided between 28 infertile patients and 28 healthy individuals (control group), all the participated individuals were aged between 18-45 years. Infertile patients with other acute or chronic diseases were excluded from the study. Serum levels of Vitamin D, TESTO, FSH, LH, E2, PRL, and total lipid were measured by standard methods. Statistical analysis has performed the results indicated statistically significant differences between the two studied groups (p≤0.05). The results showed that infertile female patients have significantly lower levels of vitamin D (p≤0.05) when compared to the healthy control groups. The results also indicated a significant increase in the level of LH and FSH (P<0.05), compared to the control group. Our study indicated a significant positive correlation between vitamin D and each of; TESTO (r=0.426, P<0.05), LH (r=0.009, P<0.01), HDL (r=0.039, P<0.05) and LH/FSH (r=0.044, P<0.05) in the serum samples of the studied infertile females, respectively.