Without longitudinal clinical data, it is difficult to differentiate some cases of chronic periodontitis (CP) and aggressive periodontitis (AgP). Furthermore, both forms of disease are exacerbated by tobacco use. Therefore, this cross-sectional study was planned, primarily, to determine the ability of Fourier-transform infrared (FTIR) spectroscopy to distinguish CP and AgP patients by analysis of human saliva samples and, secondarily, to assess the potential confounding influence of smoking on discriminating disease-specific spectral signatures. FTIR spectra were collected from patients with a clinical diagnosis of CP (n = 18; 7 smokers) or AgP (n = 23; 9 smokers). Self-reported smoking status, which may be unreliable, was confirmed by salivary cotinine analysis. Spectral band area analysis and hierarchical cluster analyses were performed to clarify if the 2 periodontitis groups as well as smoker and nonsmoker patients could be differentiated from each other. Significant variations in lipid, amino acid, lactic acid, and nucleic acid content were found between nonsmoker CP and AgP groups. Although significantly lower lipid, phospholipid, protein, amino acid, lactic acid, and nucleic acid content was noted in the smoker AgP group compared with the nonsmoker AgP group, in the CP group, phospholipid, protein, amino acid, and lactic acid content was significantly lower for smokers compared with the nonsmokers. Based on these variations, nonsmoker CP and AgP patients were discriminated from each other with high sensitivity and specificity. Successful differentiation was also obtained for the smoker CP and AgP groups. Thiocyanate levels successfully differentiated smokers from nonsmokers, irrespective of periodontal status, with 100% accuracy. Differentiation of AgP and CP forms, concomitant with determination of smoking status, may allow the dental health professional to tailor treatment accordingly.