Abstract : Oral cancer is the most common type of cancer affecting large population in India. Tobacco consumption including smoking and smokeless tobacco are some of the risk aspects for the incidence of oral cancer. Detection of premalignant oral lesions reduces the overall mortality from the disease and from the treatment procedures of oral cancer. Toluidine Blue is an acidophilic dye which has affinity for DNA and RNA present excess in dysplastic cells. In our study we have used Toluidine Blue as diagnostic test to detect premalignant oral lesions. Prospective study done in Dr. D. Y. Patil Medical College, Hospital &Research Centre, Pimpri, Pune, Under Department of General Surgery from July 2019 to August 2021. Patients presenting with oral lesions and who has risk factors for developing oral cancer were stained with an aqueous solution of 1% Toluidine blue, stained lesions were subjected to biopsy and histology. Sensitivity, Specificity and diagnostic accuracy of Toluidine Blue in detecting oral premalignant lesions among our study sample was 96.15%, 77.78%, 91% respectively. Toluidine Blue stain aids in diagnosing Premalignant Oral Lesions with good sensitivity and specificity. It is also simple, low cost and easily available.