The suppression of natural spaces due to the urban sprawl and increase of the built and agricultural environments has impacted the water resources quality, especially in areas with high population density, as the metropolitan regions. Considering the advance in Brazilian environmental legal framework, the present study aims to verify whether land use has still significantly affected water quality, through a case study in the Stones River watershed, a peri-urban river basin at a metropolitan region, Brazil. Analysis of physical-chemical indicators, collected at several sample points with different land-use (urban areas, commercial forestry, riparian forestry, mixed vegetation, pasture, and sugar cane plantation) at different seasons of the year (dry and rainy) were carried out. As a result, it was verified some statistically significant spatiotemporal effects on the of water quality caused associated to the land-use. In conclusion, in spite of the advances in the Brazilian law, land-use has still significantly affected the water quality, demanding public policies and decisions, so that effective compliance with legal guidelines is ensured.