Successful implementation of nanoparticles in biomedical applications requires surface modification by means of chiral ligands that would ensure colloidal stability of NPs in physiological media and enable them to perform chiral recognition, crucial for the majority of biological processes. Circular dichroism (CD) spectroscopy provides extensive information on the binding mode of chiral ligands to the NP-surface, as well as new characteristic properties induced by NP-ligand interaction and the nature of underlying physical phenomena. Therefore, problems of CD experiment design and interpretation of experimental data obtained constitute an essential part of any research concerned with functionalization of NPs for biomedical applications.