Abstract: The patient suffering from oral cancer usually comes to the dental surgeon when the lesion is in its advanced stages leading to poor treatment outcome and prognosis. Hence early detection is the key to success. Autofluorescence spectroscopy is a new non invasive real time technique which is explored for the early detection of oral cancer. This study has been designed to analyze the fluorescent property of tissue as it progresses from normal to malignancy when it is excited at 405 nm of light. This study uses a fiber optic-based fluorescence spectroscopy system to measure the autofluorescence spectra of six each in normal control, premalignant and malignant group of hamster`s buccal cheek pouch carcinogenesis model. The tissues were excited at 405 nm and the emission scan was obtained over a range of wavelengths. A prominent peak at 635 nm and a small peak at 700 nm in the malignant tissues when excited at 405 nm was observed which was absent in the normal and premalignant group. An addition small peak at 490 nm was observed in all the three groups. Mean intensity ratio parameters I490/635, I490/700 and I635/700 were introduced and found that the difference between the groups and within the groups was statistically significant. Further ratio parameters were able to differentiate between the normal-malignant and premalignant-malignant group. The results obtained indicate that this technique would help the surgeons to early diagnose the lesion at the early stages.