Water-soluble polymers (WSPs) are widely employed in industrial, medical, and everyday applications. Despite their widespread use, the lack of specific regulation contributes to their uncontrolled release, especially into aquatic ecosystems, raising concerns about potential risks to organisms and human health. Among WSP-containing consumer products, dishwasher pods are particularly common. These single-dose pods typically contain powder and/or liquid detergent enclosed in a water-soluble film made of polyvinyl alcohol (PVA), one of the most widely used WSPs. This study aims to investigate the potential effects of different chemical components present in PVA-based liquid dishwasher pods on Danio rerio embryos. PVA was first separated from the additives by acetone extraction and chemically characterized using ¹H-NMR spectroscopy. Embryos were then exposed to the whole pod (excluding detergent) at the concentration of 0.1 mg/L, estimated for urban wastewater entering the Milan-Nosedo treatment plant, and to the corresponding concentrations of extracted PVA (0.07 mg/L) and additives (0.03 mg/L). A multi-tier approach was adopted to investigate the effects across molecular, cellular, physiological, and organism levels, including proteomics and metabolomics analyses, reactive oxygen species (ROS) levels, acetylcholinesterase (AChE) activity, as well as mitochondrial respiration and glycolysis. Additionally, heart rate was evaluated as a physiological endpoint, while behavioural parameters (e.g. distance moved, turn angle, and thigmotaxis) were used to assess swimming behaviour. At the cellular level, ROS were significantly (p < 0.05) decreased in embryos exposed to PVA and additives compared to controls, and a significant increase in AChE activity was observed in embryos exposed to additives and pods. Additives also caused a significant decrease in the heart rate at the physiological level. Although the analysis of some last endpoints is still ongoing, these results suggest that additives in commercial PVA-based products may be the primary drivers of toxicity, consistent with previous findings in the literature.