We conducted field surveys at 25 sites in three Japanese catchments to provide conservative estimates of the safe concentration of zinc (Zn) for the protection of riverine macroinvertebrate diversity. The relationships between the Zn concentration and six macroinvertebrate metrics for taxon richness were determined by using regression analysis; this included a piecewise regression model, where two lines are joined at an unknown point. For each metric the piecewise regression model with a zero slope below a threshold concentration was selected as the best model to explain the influence of Zn. Under the assumption that macroinvertebrate diversity reductions of <10% are acceptable, the safe concentrations of Zn were estimated to be 84, 115, 84, 80, 85, and 70 µg/L for total taxon richness, Ephemeroptera, Plecoptera, and Trichoptera (EPT) richness, mayfly richness, caddisfly richness, chironomid richness, and estimated total taxon richness at the riffle scale, respectively. These concentrations are more than twice the water quality standard for Zn in Japan (30 µg/L), suggesting that the standard is likely overprotective for macroinvertebrate diversity. Field studies are useful for evaluating the level of protectiveness of safe concentrations (water quality standards) based on individual-level effects from laboratory toxicity tests, and this evaluation process will have a crucial role in implementing more purpose-driven ecological risk managements that aim to protect natural populations and communities.