Background and aims: Heavy metals pollution, particularly by hexavalent chromium, is a topic of global concern due to its high toxicity and environmental persistence. Phytoremediation emerges as a nature-based economic technology for the recovery of environments contaminated by metals. In the case of water bodies, aquatic macrophytes such as Salvinia minima Baker are excellent candidates as remedial species. To optimize its use, it is important the identification of efficient and reliable biomarkers. The objective of this work was to carry out a temporal analysis of different biochemical parameters to identify both biomarkers of exposure to Cr(VI) and the most appropriate time for their analysis in S. minima plants under laboratory conditions. M&M: Specimens of S. minima were cultured in the presence or absence of hexavalent chromium for seven days under controlled light and temperature conditions. Tissue and culture solution samples were taken at 0, 2, 4 and 7 days. The levels of H2O2, malondialdehyde, soluble and insoluble phenols and sucrose were quantified in floating pinnae and submerged rachis. Remaining Cr(VI) was determined in the treatment solution. All determinations were performed spectrophotometrically. Results: The results allowed us to identify, among the selected parameters, early (malondialdehyde), medium-term (sucrose and insoluble phenols) and late (hydrogen peroxide) biomarkers. Conclusions: The physiological implications of the observed variations are analyzed and discussed.