Osteoarthritis is emerging as the most ordinary form of arthritis, affecting 22-39% of the Indian population. A wide range of medications and therapies are available for the treatment of osteoarthritis. With a desire to develop a therapeutically effective dosage form, the present study was carried out to formulate glucosamine sulfate loaded carbopol based hydrogel. Hydrogels H1 to H6 were formulated without permeation enhancers while formulations H7 to H12 were developed with a different class of permeation enhancers such as PEG400, oleic acid, Tween 40, DMSO and PG. Based on viscosity, it was detected that formulation H4 containing polymer 1% was ideal for incorporating drug. Considering H4 as a placebo, H6 was used for further evaluation. Drug content was found to be 99.2±0.64, with in vitro drug release of 15±0.86, 22±1.59, 28±0.72, 35±0.68, 40±0.31, 47±0.83, 58±1.59, and 70±0.9 % at a duration of 1, 2, 3, 4, 5, 6, 7, 8 hours respectively. Skin irritation tests carried out on Wistar rats revealed that skin was intact with no inflammation or erythema detected, compared to untreated site. By diffusion disc method, it was evident that the levels of microbial load were relatively low, and no harmful microorganisms were identified. There were no significant changes in physicochemical properties on stability studies. Due to a simple method of preparation and effective drug delivery, glucosamine sulfate loaded hydrogels could be contemplated as a prominent formulation in the beneficiary treatment of osteoarthritis.