Determination of concentration of biomarkers of the activation of immune system, uric acid, and creatinine in the saliva can be useful tool for the diagnosis and monitoring of early manifestations of diseases such as malignant, inflammatory, and periodontal disorders. We have developed and validated a high-performance liquid chromatographic method coupled with fluorescence and diode array detection for the separation and quantification of neopterin, tryptophan, creatinine, uric acid, and kynurenine in the human saliva. A separation of these analytes was achieved within 9 min by using second-generation monolithic stationary phase and elution with phosphate buffer. The present method involves very simple sample preparation requiring small amount of sample matrix. The internal standard 3-nitro-l-tyrosine was used for a more precise quantification. The sensitivity of the present method was demonstrated with lower limits of quantification of 0.6 × 10-3 μmol/L for neopterin, 0.725 μmol/L for tryptophan, 0.12 μmol/L for creatinine, 0.18 μmol/L for uric acid, and 0.135 μmol/L for kynurenine. The method was validated with 67 real-life saliva samples collected from patients suffering from breast, ovarian, colorectal, and renal cancer, and 19 saliva samples from patients with periodontal diseases and allowed monitoring of inflammatory response.

Department of Analytical Chemistry Faculty of Pharmacy in Hradec Králové Charles University Akademika Heyrovského 1203 8 500 05 Hradec Králové Czech Republic

Department of Analytical Chemistry Faculty of Pharmacy in Hradec Králové Charles University Akademika Heyrovského 1203 8 500 05 Hradec Králové Czech Republic; Department of Clinical Biochemistry and Diagnostics University Hospital Sokolská 581 Hradec Králové 500 05 Czech Republic

Department of Dentistry Faculty of Medicine in Hradec Králové Charles University University Hospital Sokolská 581 Hradec Králové 500 05 Czech Republic

Department of Oncology Palacký University Faculty of Medicine and Dentistry Olomouc 1 P Pavlova 6 779 00 Olomouc Czech Republic

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