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The continuous release of nanoparticles and nanoplastics into the marine environment necessitates the examination of their combined effects in marine organisms. Natural Organic Matter (NOM) can significantly influence the behavior of nanomaterials in the marine environment. The present study explores the effects of algal Extracellular Polymeric Substances (EPS) in reducing the combined toxic effects of three different polystyrene nanoplastics (PSNPs)-aminated (NH 2 -PSNPs), carboxylated (COOH-PSNPs), and plain PSNPsand P25 titanium dioxide nanoparticles (Nano-TiO 2 ) towards the marine alga, Chlorella sp. Two doses (0.25 and 2.5 mg/L) of nano-TiO 2 mixed with the PSNPs (1 mg/L) were employed. The COOH-PSNPs with 2.5 mg/L nano-TiO 2 exhibited higher growth inhibition toward algal cells. Addition of algal EPS to the mixture of NMs decreased the negative effect significantly. The mean hydrodynamic diameter increased significantly from 666 to 797 nm and 1248 to 3589 nm at concentrations 0.25 and 2.5 mg/L, respectively when the mixtures of nano-TiO 2 and COOH-PSNPs were incubated with the algal EPS. In comparison to the pristine NMs, the EPS-NMs were found to significantly reduce the superoxide and hydroxyl radical production. The results were further validated with the estimation of lipid peroxidation (LPO), esterase activity, photosynthetic efficiency, and membrane permeability in the cells. The major outcomes from this study highlight the role of algal EPS in significantly reducing the toxic impact of binary mixture of NMs in marine organisms.

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