Vitamin D is a crucial fat-soluble vitamin with various functions. Its deficiency is widespread and affects individuals of all age groups. This study aimed to determine the prevalence of Vitamin D deficiency in different age groups and genders within the East Nimar region. A total of 236 participants were included, and their Vitamin D levels were measured using a Mindray CL-900i Chemiluminescence Immunoassay Analyzer. The results were stratified by age groups and gender. Among the 236 individuals, 47.9% were males and 52.1% were females. The study revealed that 86.4% of the subjects had insufficient or deficient levels of Vitamin D, while only 13.6% had sufficient levels. Specifically, 33.5% of the cases had Vitamin D deficiency, and 57.2% had Vitamin D insufficiency. Vitamin D deficiency was observed in 51.80% of individuals with higher sun exposure and 75% of those with lower exposure. Among supplement users, 45.2% had Vitamin D deficiency, whereas among non-users, the rate was 69.58%. The findings underscore the high prevalence of Vitamin D deficiency in India. Considering the inadequate dietary intake of Vitamin D in the Indian population, measures such as food fortification, Vitamin D supplementation, and exposure to sunlight (especially in the morning) are crucial to address this issue and prevent the complications associated with Vitamin D deficiency.