The new method exploits the characteristic of staphylococcal protease V8, which specifically splits peptide bonds where l-glutamic or l-aspartic acid participate. Those peptide bonds where d-aspartic acid is present remain unsplit because of the stereospecifity of enzymes. In accordance with expectations, fewer peptide bonds are split by this protease at more advanced ages and larger peptide fragments are thus formed due to the higher content of d-amino acid residues in the proteins of older people. The samples of acid-extracted non-collagenous proteins from dentin were separated using high performance liquid chromatography after enzymatic hydrolysis. A peak with a retention time of 45.3 min was chosen and his enlarging area showed a linear correlation with increasing age. Although the linear correlation with age was proved, the scattering of values decreases the usefulness of the proposed method for age estimation.