Infrared spectroscopy as a tool to monitor interactions between nanoplastics and microalgae.

The unicellular photosynthetic organisms known as microalgae are becoming one of the most important models for aquatic system studies. Among them, Chlamydomonas reinhardtii is widely used as a bioindicator of pollution or of different changes in the environment. Numerous pollutants are present in aquatic environments, particularly plastics and nanoplastics. Physiological variations after an environmental change highlight variation in the macromolecular composition of microalgae (proteins, nucleic acids, lipids and carbohydrates). Recently, Fourier transform infrared vibrational spectroscopy has been described as a reliable tool, sensitive and allowing rapid measurement of macromolecular composition of microalgae. Coupled with preprocessing and principal component analysis, it is well adapted to monitoring the effect of environmental stress on biochemical composition. In this study, infrared spectroscopy, combinedwithmultivariate analysis, has been tested first on known environmental stresses such as light intensity variation and nitrogen limitation. Then, this technique has been applied to monitor the interaction and potential impacts of polystyrene nanoparticles on microalgae. The results showed slight variations on protein and carbohydrates bands in the presence of nanoplastics, suggesting that their presence led to modifications in the biochemical composition of the microalgae. To confirm the interaction between microalgae and nanoplastics, visualization by confocal microscopy and cytotoxicity measurement has been carried out. Results showed that polystyrene nanoparticles seemed to adsorb on microalgae surface, leading to a loss of plasma membrane integrity. The resulting chemical modifications, even if moderate, could be detected by infrared spectroscopy‚ showing that this tool could be very helpful in the understanding of nanoparticlemicroalgae interaction mechanisms.