This study deals with the toxicity of the treated solutions of two types of dyes, namely, the anthraquinonic Reactive Bleu 19 dye (RB19) and the bi-azoic Direct Red 227 dye (DR227), which are treated in single and binary mixture systems. The target molecules were removed by the photocatalysis process using ZnO as a catalyst, which was calcined at two temperatures 250 and 420 °C (ZnO250 and ZnO420) prepared in the lab by the one-step calcination method. XRD, TEM, EDX, XPS, FT-IR, BET, RAMAN, and EPR analyses were carried out to characterize the catalyst material. While the phytotoxicity was being conducted using watercress seeds, the cytotoxicity took place using a cell line (raw) and an intestinal cell (caco-2). The XRD analysis showed the partial calcination of ZnO250 and the presence of anhydrous zinc acetate along with the ZnO nanoparticles (NPs). This result was not observed for ZnO420. Despite the complete discoloration (100%) of all the final solutions, ZnO250 exhibited a high cytotoxicity and phytotoxicity against the RB19 dye after the photocatalytic treatment; however, it was not the case of ZnO420 which was selected as an eco-friendly photocatalyst for the degradation of organic dyes based on the results of removal efficiency, cytotoxicity, and phytotoxicity.