Objective: Glaucoma is an eye condition characterized by optic nerve damage due to inflammation and oxidative stress, often associated with increased intraocular pressure (IOP). Vitamin D, a fat-soluble vitamin with anti-inflammatory properties, and uric acid (UA), an antioxidant molecule capable of scavenging nitrogen radicals and superoxide in plasma, were the specific biomarkers of interest in this research. Therefore, the aim of this study was to assess serum levels of vitamin D and UA in glaucoma patients within the North Indian population. Materials and methods: The study enrolled 45 age and sex-matched controls and 45 glaucoma patients. Serum levels of vitamin D and UA were measured using the radioimmunoassay (RIA) and enzymatic methods, respectively. Results: The analysis revealed that the mean of vitamin D in the test group was 23.42 ± 6.56 (ng/mL), while in the control group, it was 29.28 ± 2.58 (ng/mL). Similarly, the UA level was 4.57 ± 1.19 (mg/dL) in the test and 5.87 ± 1.74 in the control. Conclusion: The present research demonstrated that the level was significantly lowered in both vitamin D and UA in glaucoma patients compared to healthy individuals. These findings emphasize the importance of routinely estimating serum vitamin D and UA in individuals suspected of having glaucoma. Additionally, the results may prove valuable for ophthalmologists in managing and monitoring patients with glaucoma.