Purpose This study investigated whether there is a correlation between levels of 25OH-D in the follicular fluid and the serum of infertile women and the results of IVF and rates of pregnancy. The association between the levels of 25OH-D in the follicular fluid and the vitamin D repletion status was also assessed. Methods Two hundred and twenty-one infertile women participated in an IVF cycle from 2010 to 2011 in a prospective observational study. Serum and follicular fluid were collected for vitamin D analysis. Deficient, insufficient, and sufficient levels of vitamin D were defined as 10, 10-29, and 30-100 ng/ml, respectively. IVF cycle parameters and clinical pregnancy rates were also compared with the vitamin D level. Results The levels of vitamin D deficiency, insufficiency, and sufficiency were 22.6, 70.1, and 7.2 %, respectively. The fertilization rates associated with these three levels of vitamin D were 43.17, 53.37, and 58.77 %, respectively, (P = 0.054), and the implantation rates were 17.33, 15.26, and 18.75 %, respectively, (P = 0.579). No significant correlation was seen between the pregnancy rate and the serum vitamin D level (P = 0.094) or the follicular vitamin D level (P = 0.170). The serum and follicular fluid vitamin D levels showed a significant correlation (P = 0.000). Conclusion Although vitamin D is an important hormone in the human body, no correlation was found between the serum and follicular vitamin D level and the pregnancy rate in the IVF cycle.