Raman spectroscopy can provide information about the molecular composition of tissues, with potential to be applied as a diagnostic tool in lieu of histopathology. Our objectives are to determine if laser Raman spectra (RS) can be acquired reliably from the oral mucosa of patients, and to determine if the RS signature of normal oral mucosa is reproducible among anatomic oral sites and among subjects of different races and gender. 25 Caucasian and 26 Asian subjects are studied using RS with a signal acquisition time of 1 s at seven specified sites within the mouth. Multivariate analysis is used to determine the variability between tissue types and between races and gender. Unique spectra are defined for various sites in the mouth and are likely related to the degree of keratinization. However, spectral concordance by site is not greatly influenced by subject ethnicity or gender. We demonstrate, for the first time, the potential in-vivo application of RS for oral mucosal disease and demonstrate its specificity for particular mucosal types in the mouth. RS offers the potential to provide a diagnosis of disease using a noninvasive, convenient, sensitive technology that provides immediate results.