Determination of the human salivary peptides histatins 1, 3, 5 and statherin by high-performance liquid chromatography and by diode-array detection

A reversed-phase high-performance liquid chromatography (HPLC) method with diode-array detection for the quantification of several human salivary pept ides is described. Sample pretreatment consisted of the acidification of wh ole saliva by phosphate buffer. This treatment produced precipitation of mu cins, cu-amylases and other high-molecular-mass salivary proteins, simultan eous inhibition of intrinsic protease activities and reduction of sample vi scosity. Direct HPLC analysis by diode-array detection of the resulting aci dic sample allowed one to quantify histatin 1, histatin 3, histatin 5, stat herin, as well as uric acid, in normal subjects. Moreover, the groups of pe aks pertaining to proline-rich proteins and cystatins were tentatively iden tified. The method can be useful in assessing the concentration of salivary peptides from normal subjects and from patients suffering oral and/or peri odontal diseases. (C) 2001 Elsevier Science B.V. All rights reserved.