Comparison of pre-treatment methods and heavy density liquids to optimize microplastic extraction from natural marine sediments

The ubiquitous occurrence of anthropogenic particles, including microplastics in the marine environment, has, over the last years, gained worldwide attention. As a result, many methods have been developed to estimate the amount and type of microplastics in the marine environment. However, there are still no standardized protocols for how different marine matrices should be sampled or how to extract and identify these particles, making meaningful data comparison hard. Buoyant microplastics are influenced by winds and currents, and concentrations could hence be expected to be highly variable over time. However, since both high density and most of the initially buoyant microplastics are known to eventually sink and settle on the seafloor, marine sediments are proposed as a suitable matrix for microplastics monitoring. Several principles, apparatuses, and protocols for extracting microplastics from marine sediments have been presented, but extensive comparison of the different steps in the protocols using real environmental samples is lacking. Thus, in this study, different pre-treatment and subsequent density separation protocols for extraction of microplastics from replicate samples of marine sediment were compared. Two pre-treatment methods, one using inorganic chemicals (NaClO + KOH + Na(4)P(2)O(7)) and one using porcine pancreatic enzymes, as well as one with no pre-treatment of the sediment, were compared in combination with two commonly used high-density saline solutions used for density separation, sodium chloride (NaCl) and zinc chloride (ZnCl(2)). Both pre-treatment methods effectively removed organic matter, and both saline solutions extracted lighter plastic particles such as polyethylene (PE) and polypropylene (PP). The most efficient combination, chemical pre-treatment and density separation with ZnCl(2), was found to extract > 15 times more particles (≥ 100 µm) from the sediment than other treatment combinations, which could largely be explained by the high presence and efficient extraction of PVC particles.