JPI Oceans - Mikroplast - BASEMAN - NIFES - Def. basel. and standards for micropl. anal. in European waters - stand. of extr. from biota.

Increasing amounts of microplastics (MP) accumulate in marine habitats. The impact of plastic particles on aquatic ecosystems is far from understood. There is a lack of reliable data on concentrations of MP and the composition of polymers within the marine environment. Comparability of data on MP concentrations is currently hampered by the huge variety of different sampling, extraction and detection methods, making the increasing numbers of MP studies hardly -if at all- comparable. BASEMAN set out to validate and harmonize extraction and analytical methods for identification and quantification of MP. Kits of MP particles of known number, size and type in single-portion glass vials were prepared by partner UBAY (University of Bayreuth, Germany). The kits were composed of 10 particles of 4 different polymers in sizes around 1 mm, and each 50 particles of ca. 100 µm and ca. 20 µm. Institute of Marine Research (IMR, former NIFES, Bergen, Norway) has prepared and minced 9 kg soft parts of farmed blue mussel and 9 kg intestines of farmed Atlantic salmon. 9 kg intestines of wild caught haddock were cut by scissors to 3-4 cm large parts. The three biota batches were then partitioned into 100 g portions by two-increment partitioning followed by remixing to ensure homogeneity. Portions were stored frozen at -20°C in food grade glass jars with screw lids. MP-kits were blended into ¾ of biota parallel samples. All steps were carried out with precautions to avoid further MP contamination from the laboratory, equipment and solvents. Samples were de-identified and three samples with MP and one without were sent to partners for extraction and analysis of MP particle content by their available methods for recovery comparison. AWI (Alfred Wegener Institute, Germany) sent two plankton samples, caught at ?Helgoland Roads? and ?Close to Elbe?, ICBM (University Oldenburg, Germany) sent three sediment samples, silty-muddy sediment, sandy-silty sediment, and silty-muddy sediment that was heated to 550°C, spiked with MP and sent to partners as described above. Particles of ca. 20 and 100 µm: Three laboratories presented results. Recovery rates varied widely, the consortium deemed current methods as not quantitative. Main introductions of errors: 1) Number of 100µm particles added to the kit varied per polymer between 32 and 183 according to a validation test carried out on 10 kits by partner UBAY. 2) Transfer from the glass vials to the sample was not quantitative. 3) Particles loss during extraction. Conclusion: Challenge of preparing reference material and quantitative extraction is larger than the challenge of endpoint analysis. Particles around 1mm: Kits contained appropriate number of particles ±0.25% and 12 laboratories have delivered results. All added polymer types, PS, PE, PA66 and PET, were chemically identified by either FTIR or py-GC/MS by 11 partners, and counted by 12 partners. Samples with and without added plastic particles, were clearly distinguished by all partners in a blind test. No plastic fragments of that size were in procedural controls. Fibers were reported, but considered contamination. In conclusion, laboratory contamination with plastic particles in the size range of several hundred micrometers is low under the applied precautious conditions. However, also natural occurrence is low: in this case, zero in haddock and salmon intestines, heated and unheated silty-muddy sediment; 4 particles in 900 g of blue mussel soft parts, 2 particles in 8 liters of each plankton draw, 2 particles in 5 kg sandy-silty sediment. The table below indicates the number of laboratories who have delivered results, and their recovery rates as average over the three parallel samples. Matrix Number of participating laboratories recovery range [%] Blue mussel 9 95-21 Salmon intestine 7 88?56 Haddock intestine 6 91-33 Heated sediment 5 93-59 Silty-muddy sediment 5 88-38 Sandy-silty sediment 5 93-21 Both plankton samples 8 99-80 Gold standard criteria were set to general recoveries above 80% and individual polymer recoveries above 70% for biota and sediments, and to general recoveries above 90% and individual polymer recoveries above 80% for plankton samples, averaged over usually three, in few cases two parallels. For gold standard-matching protocol results, all standard deviations for the parallels for each matrix were between 0.6 and 13%. The table below indicates the number of laboratories/protocols that yielded recovery rates matching the set gold-standard criteria: Matrix Number of protocols matching gold-standard Blue mussel 3 Salmon intestine 3 Haddock intestine 2 Heated silty-muddy sediment 2 Silty-muddy sediment 1 Sandy-silty sediment 3 Plankton Helgoland Roads 3 Plankton Close to Elbe 6 The protocols with gold-standard results will be evaluated and critically discussed in a report/publication under preparation, and funneled into a protocol recommendation for MP extraction from each matrix.

We have achieved a good understanding of the quantification of larger microplastics from sediment, plankton and biota. The protocols with gold-standard results will be evaluated and critically discussed in a report/publication under preparation, and funneled into a protocol recommendation for MP extraction from each matrix. Furthermore, we have understood that currently developed methods for analysis of smaller microplastics is still not quantitative with an acceptable measurement uncertainty. Even providing reference material is challenging.

Persistent plastic litter accumulates and becomes fragmented in the marine environment over time. Together with micro-sized primary plastic litter from consumer products they lead to an increasing amount of small plastic particles (< 5 mm), so called microplastics (MP). The ubiquitous presence and massive accumulation of MP in marine habitats and the uptake of MP by various marine biota is now well recognized by scientists and authorities worldwide. However, the impact of plastic particles on aquatic ecosystems is far from understood. A fundamental issue precluding assessment of the environmental risks arising from MP is the lack of standard operation protocols (SOP) for MP sampling and detection. Consequently there is a lack of reliable data on concentrations of MP and the composition of polymers within the marine environment. Comparability of data on MP concentrations is currently hampered by a huge variety of different methods, each generating data of extremely different quality and resolution. Although MPs are recognized as an emerging contaminant in the environment, currently neither sampling, extraction, purification nor identification approaches are standardised, making the increasing numbers of MP studies hardly -if at all- comparable. BASEMAN is an interdisciplinary and international collaborative research project that aims to overcome this problem, and address the two major themes the JPI-Oceans pilot call "Ecological aspects of MP in the marine environment": 1) "The validation and harmonisation of analytical methods" which is indispensable for 2) "Identification and quantification of MP". BASEMAN's project outcomes will equip EU and national authorities with the tools and operational measures required to describe the abundance and distribution of MP in the environment. Such tools will permit evaluation of member state compliance with existing and future monitoring requirements.