Abstract
The large-scale production of plastic and the resulting release of waste is leading to a huge accumulation of micro-sized particles in the environment that could have an impact on not only aquatic organisms but also on humans. Despite the extensive literature on the subject, there is still an insufficient harmonization of methodologies for the collection and analysis of microplastics (MPs) in complex matrices; especially for high density polymers; such as polyvinyl chloride (PVC), which tend to sink and accumulate in sediments, becoming available to benthonic organisms. In this article, mussels have been chosen as model for microplastic accumulation due to their extensive filtering activity and their wide distribution in both fresh and salt water basins. To facilitate the identification and quantification of microplastics taken up by mussels, novel fluorescent and metal-doped PVC microplastics (PVC-Platinum octaethylporphyrin (PtOEP) MPs in the size range of 100 µm) have been synthesized and characterized. For the analysis of the mussels following exposure, an enzymatic protocol using amylase, lipase, papain, and SDS for organic material digestion and a sucrose-ZnCl2 density gradient for the selective separation of ingested microplastics has been developed. The final identification of MPs was performed by fluorescence microscopy. This work can greatly benefit the scientific community by providing a means to study the behavior of PVC MPs, which represent an example of a very relevant yet poorly studied high density polymeric contaminant commonly found in complex environmental matrices.