Zoledronic acid is a long-acting compound of bisphosphonate class, and it has many health and medical benefits, among which are its effects in treatment of bone disorders and accordingly decrease the incidence of osteoporosis. Besides, vitamin D is recognized as one of the fat-soluble vitamins that affect the absorption of major minerals in the digestive system. The aim of this study was to verify the influence of vitamin D on the biochemical effect of zoledronic acid. To achieve the aim of the study, the experiment was conducted on ninety-six male Wistar rats that were divided into the following groups; control group, vitamin D treated groups and zoledronic acid treated groups. The results of the study found that, taking high doses of vitamin D before treatment with zoledronic acid results in significant changes (P < 0.05) in the levels of calcium, magnesium and phosphorus, as well as levels of thyroid hormone, cortisol and levels of immunoglobulin E in the blood. Which means that treatment with zoledronic acid in high levels of vitamin D promote worsening of such biochemical parameters.