Nano-pollutants (NPLTs) have recently raised global concerns due to their possible harmful impact on environment and human health. However, until date, information on the occurrence, fate and toxicity of NPLTs in environment is scant. The knowledge gap can be attributed to the lack of advanced and sophisticated methodologies for the precise detection and characterization of NPLTs at lower concentration in complex matrices, such as surface water, wastewater, soil and food. This review briefly discusses the performance of classical methods for characterization and study of the properties of NPLTs. The important properties include shape, size, aggregation state, chemical composition and structure. Chromatographic, microscopic and spectroscopic techniques have been developed for detection and quantitative estimation of fabricated or naturally existed NPLTs in different matrices. Often, combination of these techniques is required for the separation, purification and accurate estimation. For better detection and understanding of the initial steps of interaction with the environmental matrices, pollution sources, such as wastewater and industrial discharges, must be selected as sampling points. Understanding the dynamics of agglomeration, and decantation will allow to estimate the plume of transport to delimit the potential effects.