Due to the unquestionable importance of the aquatic environment, one of the main challenges in the field of exposure science is the comprehensive characterization and evaluation of complex environmental mixtures beyond the classical/priority contaminants to new emerging contaminants [1]. The role of advanced analytical chemistry to identify and quantify potential chemical risks, that might cause adverse effects to the aquatic environment, is essential [2]. The strategies and tools that analytical chemistry has nowadays, focused on determination of various pollutants. Important challenges such as the application of effect direct analysis to reduce the complexity of the sample, the investigation of the huge number of transformation/degradation products that may be present in the soil, aquatic environment, the analysis of urban wastewater as a source of valuable information on our lifestyle [3]. At this, speech, the role of analytical chemistry and green analytical chemistry in determination of pollutants in various matrixes will be discussed.