Background: Lack of vitamin D is a serious but little-known health issue. Issue: It's not just because youngsters get rickets, including an increased risk of high blood pressure, multiple sclerosis, colon, prostate, breast, and ovarian. cancer,.and.diabetes/type 1., as. well. as Osteomalacia and osteoporosis, in, adults may have a long-term effect. The aim of study-find the correlations between. Vit.D3 to levels of hormones, imbalances (prolactin and testosterone). Subjects and methods: One hundred females were randomly selected from Baquba City, Iraq to share in this study for determination of the level of D3, prolactin, testosterone and calcium. The fluorescent immunoassay was used to detect the level of hormones and the colorimetric assay for the detection of Calcium level. Results: The result demonstrated that the level of D3 (16.75 ± 9.49) ng/mL was deficiency and effect on the level of prolactin (10.76 ± 9.49) ng/mL with positive correlation and testosterone (1.17 ± 0.37) ng/mL, leading to decrease the level of calcium (8.0 ± 2.2) mg/dL.  Conclusions: This study concluded there is a positive high correlation between the level of D3 and Prolactin] and the Testosterone hormone with age groups in females and the outcome gave evidence to the importance of the level D3 and its effect on the fertility hormones.