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Conclusions

Currently, the reliability and comparability of data on marine microplastic concentrations is hampered by the huge variety of different methodologies applied, which lead to the generation of data of extremely different quality (Hidalgo-Ruz et al. 2012). Thus, one basic aim must be the standardization of methodologies for identification and quantification of microplastics in the marine environment and the subsequent formulation of standard operating procedures (SOPs). This involves the whole cycle of the assessment of microplastics in environmental samples from sampling procedures to sample purification and identification of microplastic particles.

More research is needed in the field of representative sampling designs, especially when it comes to sampling sediments on beaches. Although sampling in itself is no challenge, choosing the appropriate sampling locations and number of replicates to representatively describe the plastic contamination of a highly dynamic environment such as a beach is indeed challenging. The suggestions by the TSG-ML (Hanke et al. 2013) are first steps towards a standardization of beach sampling within the microplastics monitoring programs of the member states of the EU.

Depending on the mesh size used, plankton or neuston nets allow for adequate sampling of a certain size class of microplastics from the water column or from surface waters. However, since nets can easily clog, especially the biologically important fraction of particles <100 µm is often severely underrepresented. Sequential size fractionated filtering of large volumes of water is a very promising technique that could provide a remedy for this problem although the applicability has yet to be proven in the field. Techniques for sampling microplastics in biological material are as variable as the biota themselves and thus a general standardization cannot be achieved. For the monitoring of microplastics in single species, such as the northern fulmar, long-term observations could lead to the establishment of standardized sampling procedures.

Sample preparation should involve a size fractionation of particles larger and smaller than 500 µm as suggested by Hidalgo-Ruz et al. (2012) (alternatively 1000 µm, TSG-ML, Hanke et al. 2013) to enable comparability of data from different studies including studies that focus on particles <500 µm. Currently, no SOP exists for extraction solutions and techniques used to extract microplastic particles from samples. The use of extraction solutions of lower densities (e.g. NaCl) results in underestimation of microplastics of higher densities. Zinc chloride is a relatively cheap and recyclable solution that can be used to produce high-density solutions, which extract the whole spectrum of common user plastics. Furthermore, we suggest using separation techniques with high recovery rates in the whole size spectrum of microplastic particles at low temporal and personal effort such as the Microplastic Sediment Separator (Imhof et al. 2012).

Identification by spectroscopy is essential, especially for particles <500 µm. The quality of the spectroscopic results relies strongly on effective sample purification. Purifying steps with agents that negatively affect certain plastic polymers (e.g. strong acidic or alkaline solutions) should be avoided to conserve the whole spectrum of plastic contamination within an environmental sample. Gentle approaches such as sequential enzymatic digestion have been successfully applied by the authors and provide efficient purification for subsequent spectroscopic analysis. The contamination of microplastic samples during the whole assessment procedure from sampling to analysis is a highly urgent topic that needs to be addressed by standardised contamination prevention techniques. In this context, the avoidance of plastic material as far as possible as well as the use of control samples should be implemented as key elements of microplastics investigations.

An example of recent research by the authors showed that the commonly applied visual inspection of samples alone is insufficient to identify microplastic particles in environmental samples. This is especially the case for particles

<500 µm, which have to be concentrated on filters for analysis. In this context, a spectroscopic verification of microplastic particles by micro-Raman or microFTIR spectroscopy is essential and has also been used successfully for analyzing marine samples. Spectroscopic techniques create an added value as they also provide information on the polymer composition. Although FTIR spectroscopy has been used more frequently in microplastics research (Hidalgo-Ruz et al. 2012) Raman spectroscopy is of equal value for analyzing microplastic samples.

By combining spectroscopic techniques with fast area-resolved measurements, with e.g. FPA detectors, and chemical imaging, it is possible to scan whole sample filters in a fraction of time compared to chemical mapping with single detectors (Harrison et al. 2012). However, SOPs for the use of this highly promising novel techniques are not accessible at present but will soon be published (Löder et al. 2015). The standardized use of spectroscopy in microplastic research finally enables the generation of valid data on concentrations, particle size distribution, involved polymers and distribution among different marine habitats and in marine biota. This is essential to guarantee comparability of different data sets. Furthermore, the data generated will serve as basis for the design of realistic laboratory experiments on the impact of microplastics on marine biota. These steps are crucial for a reliable evaluation and the assessment of potential impacts and risks of microplastics in the world's oceans.

Acknowledgments The authors would like to thank the German Federal Ministry of Education and Research (BMBF) and the Alfred-Wegener-Institut Helmholtz-Zentrum für Polar- und Meeresforschung (AWI) for funding the project MICROPLAST. Furthermore, we would like to thank the NLWKN for preparation and providing the microplastics samples for the case study and Dr. Sonja Oberbeckmann for providing the manta trawl picture. This is publication number 37791 of the AlfredWegener-Institut Helmholtz-Zentrum für Polar- und Meeresforschung.

 
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