The use of engineered nanoparticles in the environment and human life has increased in the last 20 years. The risk assessment concerning application of nanomaterials in biological systems requires their thorough characterization. Understanding the correlations between physicochemical properties of nanoparticles concerning not only the size, particle size distribution, number concentration, degree of aggregation, or agglomeration but also solubility, stability, binding affinity, surface activity, chemical composition, and nanoparticle synthesis yield allows their reliable characterization. Thus, to find the structure-function/property relationship of nanoparticles, multifaceted characterization approach based on more than one analytical technique is required. On the other hand, the increasing demand for identification and characterization of nanomaterials has contributed to the continuous development of spectrometric techniques which enables for their qualitative and quantitative analysis in complex matrices giving reproducible and reliable results. This review is aimed at providing a discussion concerning four main aspects of nanoparticle characterization: nanoparticle synthesis yield, particle size and number concentration, elemental and isotopic composition of nanoparticles, and their surface properties. The conventional and non-conventional spectrometric techniques such as spectrophotometry UV-Vis, mass spectrometric techniques working in conventional and single-particle mode, or those based on optical emission detection systems are described with special emphasis paid on their advantages and drawbacks. The application and recent advances of these methods are also comprehensively reviewed and critically discussed.Graphical abstract