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Optimization of an Analytical Protocol for the Extraction of Microplastics from Seafood Samples with Different Levels of Fat †

Diogo M. Silva
Marisa Almeida
Francisco Guardiola
Sabrina M. Rodrigues
1,2 and
Sandra Ramos
ICBAS—Institute of Biomedical Sciences Abel Salazar, Porto University, Rua de Jorge Viterbo Ferreira n° 228, 4050-313 Porto, Portugal
CIIMAR—Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Terminal de Cruzeiros do Porto de Leixões, Avenida General Norton de Matos, S/N, 4450-208 Matosinhos, Portugal
Faculty of Biology, C. Campus Universitario, University of Murcia, 5, 30100 Murcia, Spain
Author to whom correspondence should be addressed.
Presented at the IX Iberian Congress of Ichthyology, Porto, Portugal, 20–23 June 2022.
Presenting author (poster).
Biol. Life Sci. Forum 2022, 13(1), 75;
Published: 9 June 2022
(This article belongs to the Proceedings of The IX Iberian Congress of Ichthyology)


The global production of plastics increased from 1.5 million tons in 1950 to 370 million tons in 2020. Microplastics (MPs) are plastic particles smaller than 5 mm, with a concerning ubiquitous occurrence in marine environments, posing risks to wildlife and humans. Seafood is an important source of proteins (including all the essential amino acids), and other essential nutrients with numerous health benefits. Human exposure to MPs through the consumption of contaminated seafood, together with the potential of these particles to cause health risks, is motivating a better understanding of the security of our diets. In recent years, several methodologies have been used to investigate MP occurrence in seafood, although the existence of complex sample matrices, namely, a high level of fats, can pose severe difficulties and compromise the efficiency of MP quantification. To solve this issue, the present study aimed to develop a detailed protocol suitable to process seafood samples with different levels of fats (fish and mollusks, from fresh and canned sources). For the sample digestion, several tests were performed using two solutions (10% KOH, 30% H2O2) with different volumes, temperatures (40 °C, 65 °C) and durations (24, 48, 72 h) of incubation. To remove the fat remaining after digestion, three detergents (two laboratory surfactants and a commercial washing-up liquid) and 96% ethanol were tested. Manual recovery of the fat layer was also tested. For filtration, two filter membranes were compared (glass microfiber and nitrate cellulose filters, 0.45 µm pore size). The efficiency of the different experiments was determined through the observation and estimation, in percentage, of the organic matter digestion and post-digestion fat removal. The methodology optimized in this study combined a sample digestion with 30% H2O2 incubated at 65 °C, for 24 to 48 h, with a manual separation of the post-digestion fats with immediate observation in a stereomicroscope. After, this methodology was applied to different types of polymers (e.g., polyethylene, polypropylene, polyethylene-terephthalate, polystyrene), to investigate if these procedures altered the integrity of MPs. The results show that this methodology will allow us to efficiently process complex seafood samples with different fat levels, without compromising MPs’ integrity.


This research was partially funded by Ocean3R (NORTE-01-0145-FEDER-000064) and ATLANTIDA (ref. NORTE-01-0145-FEDER-000040) projects, supported by the Norte Portugal Regional Operational Programme (NORTE 2020) under the PORTUGAL 2020 Partnership Agreement and through the European Regional Development Fund (ERDF). Also, FCT is awarded by the Strategic Funding UIDB/04423/2020 and UIDP/04423/2020 through national funds provided by FCT and ERDF, a PhD fellowship to DS (2020.06088.BD) and a research contract to SRamos (DL57/2016/CP1344/CT0020).

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Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.
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MDPI and ACS Style

Silva, D.M.; Almeida, M.; Guardiola, F.; Rodrigues, S.M.; Ramos, S. Optimization of an Analytical Protocol for the Extraction of Microplastics from Seafood Samples with Different Levels of Fat. Biol. Life Sci. Forum 2022, 13, 75.

AMA Style

Silva DM, Almeida M, Guardiola F, Rodrigues SM, Ramos S. Optimization of an Analytical Protocol for the Extraction of Microplastics from Seafood Samples with Different Levels of Fat. Biology and Life Sciences Forum. 2022; 13(1):75.

Chicago/Turabian Style

Silva, Diogo M., Marisa Almeida, Francisco Guardiola, Sabrina M. Rodrigues, and Sandra Ramos. 2022. "Optimization of an Analytical Protocol for the Extraction of Microplastics from Seafood Samples with Different Levels of Fat" Biology and Life Sciences Forum 13, no. 1: 75.

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