Introduction: Vitamin D plays a significant role in the functioning of the immune system and it influences many dermatological diseases such as psoriasis and atopic dermatitis. The prevalence of vitamin D deficiency is growing globally with around 30–50% of people are known to have low levels of vitamin D. Acne vulgaris is a common inflammatory disorder of the pilosebaceous unit. Studies about the role of vitamin D in the pathogenesis of acne vulgaris have shown conflicting and nonconclusive results. Thus, the precise purpose of vitamin D has not yet been established. Aims & Objective: To evaluate serum levels of vitamin D in sample of patients with acne vulgaris and compare it with matched healthy controls & to investigate if there is any relation between serum vitamin D level and the severity of acne vulgaris. Materials and Methods: This cross-sectional study included 40 patients with acne vulgaris and 40 matched healthy controls. Serum 25-hydroxyvitamin D [25 (OH) D] levels were measured for both patients and healthy controls. Results: Patients with acne are younger than healthy controls and female preponderance was observed as compared with male patients. The mean (± SD) concentrations of 25[OH]D in patient group and the control group were 21.3 ± 9.7 ng mL and 29.6 ± 11.2 ng/mL respectively. The difference was found to be statistically significant (P < 0.05). Thus our study yielded lower levels of serum 25-hydroxyvitamin D in patients with acne vulgaris than its level in healthy controls. Conclusion: This study has shown clearly that in patients with acne Vulgaris vitamin D deficiency is more evident with P-value P < 0.05. Moderate to severe vitamin deficiency was observed in 27 cases (67.5%) and in 20 controls (50%) (P < 0.05). Further clinical studies with larger sample size are required to address the importance of vitamin D in pathogenesis & grading of Acne vulgaris. Specifically, to determine the effects of treatment of acne with both topical vitamin D analogs and vitamin D supplementation.