A three-stage sequential extraction procedure optimized by the European Community Bureau of Reference (BCR) was applied to assess the geochemical behavior and environmental mobility of As, Cr, Cu, Ni, Pb and Zn in surface sediments from two ecosystems under different anthropogenic influences, the Estuarine Complex of Santos and São Vicente (ECSSV) upper estuary (high anthropogenic influence) and the Bertioga channel (low anthropogenic influence), both of which are located on the coast of São Paulo, Brazil. In general, the difference between the regions varied considerably for almost all these elements, and the values for the Bertioga channel were always lower than those found in the upper estuary. Among all the elements, Zn showed the greatest mobility, being present mainly in the acid-soluble fraction. Arsenic and Pb were associated with the reducible fraction, and Cu and Ni in the oxidizable fraction. Meanwhile, Cr was mainly found in the residual fraction. Although previous studies, regarding total concentrations suggest the probable enrichment of Cu, Pb and Zn in the surface sediments of the ECSSV, according to our results and the Risk Assessment Code (RAC) analysis, only Zn presented a high risk of bioavailability, and the modified Risk Assessment Code (mRAC) only showed six samples in the inner estuary with a medium risk of mobilization. Therefore, this study can serve as a management tool and optimize the monitoring efforts of chemical pollution in the region. Keywords Estuarine complex of santos and são vicente • Potentially toxic elements • Sequential extraction • Risk assessment code (RAC) due to its strategic location between Santos Port and São Paulo city. The area hosts the largest industrial complex in the country (Cubatão Industrial Complex). It is already considered by many studies to be environmentally affected by industrial, urban and tourist activities, indicated mainly by contamination through potential harmful elements/compounds, like metals and organic compounds (e.g., Bordon et al. 2011; Martins et al. 2011; Buruaem et al. 2013; Kim et al. 2019) and effects on the biological community and mangroves (e.g., Cordeiro and Costa 2010; Cesar et al. 2014; Damasio et al. 2020). These activities have developed over the past century, and the increasing anthropogenic influence affects the resilience of the estuary (Buruaem et al. 2013; Cesar et al. 2014). However, while many studies have been conducted in the region that assess the levels of trace elements (e.g., Silva et al. 2011; Torres et al. 2015; Kim et al. 2016), studies on the environmental availability and geochemical behavior of these Communicated by M. V. Alves Martins