Rapid development in medicine and pharmacy has created a need for novel biomaterials with advanced properties such as photoluminescence, biocompability and long-term stability. The following research deals with the preparation of novel types of N-doped chitosan-based carbon quantum dots. Nanomaterials were obtained with simultaneous nitrogen-doping using biocompatible amino acids according to Green Chemistry principles. For the carbon quantum dots synthesis chitosan was used as a raw material known for its biocompability. The nanomaterials obtained in the form of lyophilic colloids were characterized by spectroscopic and spectrofluorimetric methods. Their quantum yields were determined. Additionally the cytotoxicity of the prepared bionanomaterials was evaluated by XTT (2,3-Bis-(2-methoxy-4-nitro5-sulfophenyl)-2H-tetrazolium-5-carboxanflide salt) method. Our results confirmed the formation of biocompatible quantum dots with carbon cores exhibiting luminescence in visible range. Performed studies showed that modification with lysine (11.5%) and glutamic acid (7.4%) had a high impact on quantum yield, whereas functionalization with amino acids rich in S and N atoms did not significantly increase in fluorescence properties. XTT assays as well as morphological studies on human dermal fibroblasts confirmed the lack of cytotoxicity of the prepared bionanomaterials. The study shows chitosan-based quantum dots to be promising for biomedical applications such as cell labelling, diagnostics or controlled drug delivery and release systems.Web of Science92art. no. 27