Fig. 2

Experimental workflow of mixture analysis, exemplarily shown for mixC.1. Single substance toxicity observation. a Exposure of ZFE to different concentrations of single substances and effect determination at different time points. b Single substance concentration–response curve (ss_CRC) for all mixture components (triangle: carbendazim, circle: cyprodinil, diamond: diclofenac, star: diuron, square: genistein) obtained after an exposure duration of 24 hpe (pink) and 48 hpe (purple). Mixture toxicity prediction based on ss_CRCs (black) with the CA (light blue) and the IA (yellow) model, respectively, for the exposure durations of 24 hpe (c, pink), and 48 hpe (d, purple). e Exposure of ZFE to different concentrations of the mixture (fixed MR) and effect determination at different time points. f Observed mixture toxicity (mix_CRC) at two time points. Comparison of observed (black) and predicted mixture toxicity (light blue: CA, yellow: IA) for the 24 hpe (g, pink) and 48 hpe (h, purple) exposure duration