Towards a reference material for microplastics' number concentration-case study of PET in water using Raman microspectroscopy.

Increasing demand for size-resolved identification and quantification of microplastic particles in drinking water and environmental samples requires the adequate validation of methods and techniques that can be used for this purpose. In turn, the feasibility of such validation depends on the existence of suitable certified reference materials (CRM). A new candidate reference material (RM), consisting of polyethylene terephthalate (PET) particles and a water matrix, has been developed. Here, we examine its suitability with respect to a homogeneous and stable microplastic particle number concentration across its individual units. A measurement series employing tailor-made software for automated counting and analysis of particles (TUM-ParticleTyper 2) coupled with Raman microspectroscopy showed evidence of the candidate RM homogeneity with a relative standard deviation of 12% of PET particle counts involving particle sizes >30 µm. Both the total particle count and the respective sums within distinct size classes were comparable in all selected candidate RM units. We demonstrate the feasibility of production of a reference material that is sufficiently homogeneous and stable with respect to the particle number concentration.

Microplastics in the bogue, Boops boops: A snapshot of the past from the southern Tyrrhenian Sea.

The present investigation focuses on Boops boops specimens gathered in the Gulf of Patti in 2010. Providing a snapshot from the past, this paper represents, chronologically, the first record of microplastic ingestion in the Mediterranean bogue. The plastic abundance and composition in gastrointestinal tracts of the bogue was assess, in order to improve the knowledge on spatial-temporal variability of microplastics pollution in the Mediterranean basin and in particular, in the southern Tyrrhenian Sea. In a total of 65 specimens, 180 particles of plastic (2.8 items/specimens), mainly belonging to microplastics class, were found. Fragments (63%) and fibres (30%) were the predominant shape categories. Eleven polymers were identified: polypropylene and polyethylene were the most abundant. Several synthetic polymers belonging to the class of elastomers were also observed. The study area is strongly influenced by the absence of trawl fishing activities and a low mixing level of the seabed that, together with the confluence of different watercourses and the presence of different kind of anthropic impact, including motorway, could make it a 'waste disposal site'. Finally, our results suggest the usefulness to retrieve older samples to better understand spatial-temporal changes in marine litter pollution over time.

A new digestion approach for the extraction of microplastics from gastrointestinal tracts (GITs) of the common dolphinfish (Coryphaena hippurus) from the western Mediterranean Sea.

Plastic ingestion is one of the main impacts of marine litter on organisms. The occurrence of microplastics (MPs < 5 mm) in the stomachs of Mediterranean species was already reported in several studies. In this context, the present study aims to develop a new approach of digestion for the identification of MPs in the gastrointestinal tracts (GITs) of marine organisms. The new approach combines two digestion protocols, including potassium hydroxide (KOH) and nitric acid (HNO3), to remove most organic and inorganic materials. This digestion allows recording small MPs that are difficult to find via routinely stomach content analysis and also to minimize the overestimation of the phenomenon trough the control of airborne contamination. The new approach was tested on a voracious pelagic opportunistic predator, the common dolphinfish, a fishery resource exploited in several Mediterranean areas. The results showed that a large amount of ingested meso- and microplastics, such as fragments or sheets, was recorded in GITs (F = 65.5 %). The FTIR analysis on litter samples allowed to identify polyethylene, polypropylene and polystyrene as dominant constituent polymers of microplastics. These results confirmed that our novel combined digestion protocol represents a reliable approach to detect MPs in opportunistic pelagic predators.

Riverine anthropogenic litter load to the Mediterranean Sea near the metropolitan area of Barcelona, Spain.

Every year >4 million tonnes of plastic are estimated to enter the oceans and much of it comes from land-based sources through rivers and estuaries. To fill the lack of information related to plastic inputs from rivers, a harmonized approach based on visual observations for monitoring floating macro litter was followed in this work. We provide the results of one-year monitoring (October 2016-September 2017) in the Llobregat and El Besòs rivers, which are flowing through an industrialized and populated area nearby the city of Barcelona (Catalonia, NE Spain). Floating litter items categories were influenced by urban centres located along the rivers. Overall, similar litter composition was observed in both rivers with a prevalence of plastics, mainly related to the food and beverage sectors. Seasonal variability showed significant correlations with natural factors such as wind and rainfall. Approximately 0.4-0.6 tonnes of plastic per year were estimated to be loaded into the sea by these two Catalan rivers. This study contributes to enlarge our knowledge on anthropogenic riverine litter entering the NW Mediterranean Sea, providing a starting point for the development of further mitigation strategies.

Trace analysis of polystyrene microplastics in natural waters.

The development of quantitative and qualitative analytical methods to assess micro-plastics (MPLs) and nano-plastics (NPLs) content in the environment is a central issue for realistic risk assessment studies. However, the quantitative analysis continues being a critical issue, in particular for MPLs from 100 µm down to the nano-sized range in complex environmental samples. This paper evaluates the potential of mass spectrometry for the analysis of MPLs and NPLs. The performance of different techniques including matrix-assisted laser desorption ionisation (MALDI) coupled to time-of-flight mass spectrometry (TOF-MS), liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS), and the ambient ionisation approaches as desorption electrospray ionisation (DESI) and direct analysis real-time (DART), were assessed for the study of polystyrene (PS) MPLs and NPLs in natural waters. A method based on LC-HRMS, equipped with an atmospheric pressure photoionisation source (APPI), operated in negative conditions for the quantitative analysis of PS MPLs and NPLs in natural waters, was developed. The chromatographic separation was achieved using an advanced polymer chromatographic (APC) column using toluene isocratic as the mobile phase. The optimal analytical method showed an instrumental limit of detection (ILOD) of 20 pg and methods limits of detection and quantification around 30 pg L-1 and 100 pg L-1, respectively. And, recoveries of 60 and 70% in samples from rivers and the marine coast, respectively. The performance of the new method was proved by the analysis of fortified samples and natural seawater samples.

Metabolic Responses of Mytilus galloprovincialis to Fullerenes in Mesocosm Exposure Experiments.

In this study, Mediterranean mussels (Mytilus galloprovincialis) were exposed through the diet to fullerene soot at three concentrations in parallel to a control group. Their metabolomics response was assessed by high-performance liquid chromatography coupled to high-resolution mass spectrometry (HPLC-HRMS). The experiments were conducted in marine mesocosms, during 35 days (7 days of acclimatization, 21 days of exposure, and 7 days of depuration). Real conditions were emulated in terms of physicochemical conditions of the habitat. Results confirmed the bioaccumulation of fullerenes, and the metabolome of the exposed organisms revealed significant differences in the concentrations of seven free amino acids in comparison to the control group. An increase in small nonpolar amino acids (e.g., alanine) and branched chain amino acids (leucine and isoleucine) were observed. Also, glutamine concentrations decreased significantly, suggesting the activation of facultative anaerobic energy metabolism. Branched chain amino acids, such as leucine and isoleucine, followed the opposite trend after the highest level of exposure, which can imply hormesis effects. Other significant differences were observed on lipids content, such as the general increase of free fatty acids, i.e., long-chain fatty acids (lauric, myristic, and palmitic acids) when the concentration of exposure was increased. These results were consistent with hypoxia and oxidative stress.

Cytotoxic effects of commonly used nanomaterials and microplastics on cerebral and epithelial human cells.

Plastic wastes are among the major inputs of detritus into aquatic ecosystems. Also, during recent years the increasing use of new materials such as nanomaterials (NMs) in industrial and household applications has contributed to the complexity of waste mixtures in aquatic systems. The current effects and the synergism and antagonisms of mixtures of microplastics (MPLs), NMs and organic compounds on the environment and in human health have, to date, not been well understood but instead they are a cause for general concern. The aim of this work is to contribute to a better understanding of the cytotoxicity of NMs and microplastics/nanoplastics (MPLs/NPLs), at cell level in terms of oxidative stress (evaluating Reactive Oxygen Species effect) and cell viability. Firstly, the individual cytotoxicity of metal nanoparticles (NPs) (AgNPs and AuNPs), of metal oxide NPs (ZrO2NPs, CeO2NPs, TiO2NPs, and Al2O3NPs), carbon nanomaterials (C60fullerene, graphene), and MPLs of polyethylene (PE) and polystyrene (PS) has been evaluated in vitro. Two different cellular lines T98G and HeLa, cerebral and epithelial human cells, respectively, were employed. The cells were exposed during 24-48h to different levels of contaminants, from 10ng/mL to 10µg/mL, under the same conditions. Secondly, the synergistic and antagonistic relationships between fullerenes and other organic contaminants, including an organophosphate insecticide (malathion), a surfactant (sodium dodecylbenzenesulfonate) and a plasticiser (diethyl phthalate) were assessed. The obtained results confirm that oxidative stress is one of the mechanisms of cytotoxicity at cell level, as has been observed for both cell lines and contributes to the current knowledge of the effects of NMs and MPLs-NPLs.