Effect of micro- and nanoplastic particles on human macrophages.

Micro- and nanoplastics (MNPs) are ubiquitous in the environment, resulting in the uptake of MNPs by a variety of organisms, including humans, leading to particle-cell interaction. Human macrophages derived from THP-1 cell lines take up Polystyrene (PS), a widespread plastic. The question therefore arises whether primary human macrophages also take up PS micro- and nanobeads (MNBs) and how they react to this stimulation. Major aim of this study is to visualize this uptake and to validate the isolation of macrophages from peripheral blood mononuclear cells (PBMCs) to assess the impact of MNPs on human macrophages. Uptake of macrophages from THP-1 cell lines and PBMCs was examined by transmission electron microscopy (TEM), scanning electron microscopy and live cell imaging. In addition, the reaction of the macrophages was analyzed in terms of metabolic activity, cytotoxicity, production of reactive oxygen species (ROS) and macrophage polarization. This study is the first to visualize PS MNBs in primary human cells using TEM and live cell imaging. Metabolic activity was size- and concentration-dependent, necrosis and ROS were increased. The methods demonstrated in this study outline an approach to assess the influence of MNP exposure on human macrophages and help investigating the consequences of worldwide plastic pollution.

Size dependent uptake and trophic transfer of polystyrene microplastics in unicellular freshwater eukaryotes.

Microplastics (MP) have become a well-known and widely investigated environmental pollutant. Despite the huge amount of new studies investigating the potential threat posed by MP, the possible uptake and trophic transfer in lower trophic levels of freshwater ecosystems remains understudied. This study aims to investigate the internalization and potential trophic transfer of fluorescent polystyrene (PS) beads (0.5 µm, 3.6 × 108 particles/mL; 6 µm, 2.1 × 105 particles/mL) and fragments (<30 µm, 5 × 103 particles/mL) in three unicellular eukaryotes. This study focuses on the size-dependent uptake of MP by two freshwater Ciliophora, Tetrahymena pyriformis, Paramecium caudatum and one Amoebozoa, Amoeba proteus, serving also as predator for experiments on potential trophic transfer. Size-dependent uptake of MP in all three unicellular eukaryotes was shown. P. caudatum is able to take up MP fragments up to 27.7 µm, while T. pyriformis ingests particles up to 10 µm. In A. proteus, small MP (PS0.5µm and PS6µm) were taken up via pinocytosis and were detected in the cytoplasm for up to 14 days after exposure. Large PS-MP (PS<30µm) were detected in A. proteus only after predation on MP-fed Ciliophora. These results indicate that A. proteus ingests larger MP via predation on Ciliophora (PS<30µm), which would not be taken up otherwise. This study shows trophic transfer of MP at the base of the aquatic food web and serves as basis to study the impact of MP in freshwater ecosystems.

Identification of different plastic types and natural materials from terrestrial environments using fluorescence lifetime imaging microscopy.

Environmental pollution by plastics is a global issue of increasing concern. However, microplastic analysis in complex environmental matrices, such as soil samples, remains an analytical challenge. Destructive mass-based methods for microplastic analysis do not determine plastics' shape and size, which are essential parameters for reliable ecological risk assessment. By contrast, nondestructive particle-based methods produce such data but require elaborate, time-consuming sample preparation. Thus, time-efficient and reliable methods for microplastic analysis are needed. The present study explored the potential of frequency-domain fluorescence lifetime imaging microscopy (FD-FLIM) for rapidly and reliably identifying as well as differentiating plastics and natural materials from terrestrial environments. We investigated the fluorescence spectra of ten natural materials from terrestrial environments, tire wear particles, and eleven different transparent plastic granulates <5 mm to determine the optimal excitation wavelength for identification and differentiation via FD-FLIM under laboratory conditions. Our comparison of different excitation wavelengths showed that 445 nm excitation exhibited the highest fluorescence intensities. 445 nm excitation was also superior for identifying plastic types and distinguishing them from natural materials from terrestrial environments with a high probability using FD-FLIM. We could demonstrate that FD-FLIM analysis has the potential to contribute to a streamlined and time-efficient direct analysis of microplastic contamination. However, further investigations on size-, shape-, color-, and material-type detection limitations are necessary to evaluate if the direct identification of terrestrial environmental samples of relatively low complexity, such as a surface inspection soil, is possible.

The cuticle proteome of a planktonic crustacean.

The cuticles of arthropods provide an interface between the organism and its environment. Thus, the cuticle's structure influences how the organism responds to and interacts with its surroundings. Here, we used label-free quantification proteomics to provide a proteome of the moulted cuticle of the aquatic crustacean Daphnia magna, which has long been a prominent subject of studies on ecology, evolution, and developmental biology. We detected a total of 278 high-confidence proteins. Using protein sequence domain and functional enrichment analyses, we identified chitin-binding structural proteins and chitin-modifying enzymes as the most abundant protein groups in the cuticle proteome. Structural cuticular protein families showed a similar distribution to those found in other arthropods and indicated proteins responsible for the soft and flexible structure of the Daphnia cuticle. Finally, cuticle protein genes were also clustered as tandem gene arrays in the D. magna genome. The cuticle proteome presented here will be a valuable resource to the Daphnia research community, informing genome annotations and investigations on diverse topics such as the genetic basis of interactions with predators and parasites.

Multi-method analysis of microplastic distribution by flood frequency and local topography in Rhine floodplains.

Rivers are important transport pathways for microplastics into the ocean, but they can also be potential sinks due to microplastic deposition in the sediments of the river bed and adjacent floodplains. In particular, floods can (re)mobilise microplastics from sediments and floodplains, (re)deposit and relocate them depending on the floodplain topography. The knowledge about fluvial microplastic input to floodplains, their spatial distribution and their fate in floodplain soils is limited. To investigate this topic, we sampled soil at a depth of 5-20 cm along three transects in three different Rhine floodplains. We analysed the soil samples in tandem with pyrolysis GC/MS and ATR- & µ-FPA-FTIR for their microplastic abundance and mass concentrations. To study the influence of flood frequency on the microplastic abundance in the three floodplains, we fitted a hydrodynamic flood model (MIKE 21, DHI, Hørsholm, Denmark) and related the results to the respective spatial microplastic distribution. We found similar microplastic distribution patterns in each floodplain. The highest microplastic abundance (8516-70,124 microplastics kg-1) and mass concentration (46.2-141.6 mg kg-1) were consistently found in the farthest transects from the Rhine in a topographical depression. This microplastic distribution pattern is detectable with both, pyrolysis GC/MS and FTIR. The strongest correlation between the results of both methods was found for small, abundant microplastic particles. Our results suggest that the spatial distribution of microplastics in floodplains is related to the combination of flood frequency and local topography, that ought to be explicitly considered in future studies conducted in floodplains. Finally, our results indicate that pyrolysis GC/MS and FTIR data are comparable under certain conditions, which may help in the decision for the analytical method and sampling design in future studies.

Unraveling root and rhizosphere traits in temperate maize landraces and modern cultivars: Implications for soil resource acquisition and drought adaptation.

A holistic understanding of plant strategies to acquire soil resources is pivotal in achieving sustainable food security. However, we lack knowledge about variety-specific root and rhizosphere traits for resource acquisition, their plasticity and adaptation to drought. We conducted a greenhouse experiment to phenotype root and rhizosphere traits (mean root diameter [Root D], specific root length [SRL], root tissue density, root nitrogen content, specific rhizosheath mass [SRM], arbuscular mycorrhizal fungi [AMF] colonization) of 16 landraces and 22 modern cultivars of temperate maize (Zea mays L.). Our results demonstrate that landraces and modern cultivars diverge in their root and rhizosphere traits. Although landraces follow a 'do-it-yourself' strategy with high SRLs, modern cultivars exhibit an 'outsourcing' strategy with increased mean Root Ds and a tendency towards increased root colonization by AMF. We further identified that SRM indicates an 'outsourcing' strategy. Additionally, landraces were more drought-responsive compared to modern cultivars based on multitrait response indices. We suggest that breeding leads to distinct resource acquisition strategies between temperate maize varieties. Future breeding efforts should increasingly target root and rhizosphere economics, with SRM serving as a valuable proxy for identifying varieties employing an outsourcing resource acquisition strategy.

Sexual dimorphism in the proventriculus of the buff-tailed bumblebee Bombus terrestris (L. 1758) (Hymenoptera: Apidae).

Research on eusocial bee species like Bombus terrestris is primarily focused on the worker caste, which is why their morphology and anatomy are already well described. This includes the alimentary tract, which is adapted for feeding on nectar and pollen. Located at the transition between crop and ventriculus is a highly specialised compartment, the proventriculus. In female workers of B. terrestris, the proventriculus is surrounded by muscles and consists of four anterior lips. A detailed description, however, is only provided for B. terrestis worker bees while studies on the proventriculus of the male reproductive caste are absent. Here, we provide a detailed analysis of the differences between the proventriculus of the B. terrestris males and females through morphometrics, histology and scanning electron microscopy imaging, and unravel a distinct sexual dimorphism. The male proventriculus is wider resulting in a greater volume than the female proventriculus. Histological analysis revealed 4 distinctive chambers of the male proventriculus, which are completely covered with hairs on the inside. In contrast, those chambers in the proventriculus of female B. terrestris, are only rudimentarily present forming only small pouches with hairs in the junctions between the proventricular folds inside the proventriculus. The morphological differences in the proventriculus may be based on different modi vivendi, as males do not return to the colony and fly longer distances. This and the synthesis of sperm and mating plug might require higher energy reserves, leading to the necessity of higher food storage capacities.

A promising method for fast identification of microplastic particles in environmental samples: A pilot study using fluorescence lifetime imaging microscopy.

Microplastic pollution of the environment has been extensively studied, with recent studies focusing on the prevalence of microplastics in the environment and their effects on various organisms. Identification methods that simplify the extraction and analysis process to the point where the extraction can be omitted are being investigated, thus enabling the direct identification of microplastic particles. Currently, microplastic samples from environmental matrices can only be identified using time-consuming extraction, sample processing, and analytical methods. Various spectroscopic methods are currently employed, such as micro Fourier-transform infrared, attenuated total reflectance, and micro Raman spectroscopy. However, microplastics in environmental matrices cannot be directly identified using these spectroscopic methods. Investigations using frequency-domain fluorescence lifetime imaging microscopy (FD-FLIM) to identify and differentiate plastics from environmental materials have yielded promising results for directly identifying microplastics in an environmental matrix. Herein, two artificially prepared environmental matrices that included natural soil, grass, wood, and high-density polyethylene were investigated using FD-FLIM. Our first results showed that we successfully identified one plastic type in the two artificially prepared matrices using FD-FLIM. However, further research must be conducted to improve the FD-FLIM method and explore its limitations for directly identifying microplastics in environmental samples.

Nominally identical microplastic models differ greatly in their particle-cell interactions.

Due to the abundance of microplastics in the environment, research about its possible adverse effects is increasing exponentially. Most studies investigating the effect of microplastics on cells still rely on commercially available polystyrene microspheres. However, the choice of these model microplastic particles can affect the outcome of the studies, as even nominally identical model microplastics may interact differently with cells due to different surface properties such as the surface charge. Here, we show that nominally identical polystyrene microspheres from eight different manufacturers significantly differ in their ζ-potential, which is the electrical potential of a particle in a medium at its slipping plane. The ζ-potential of the polystyrene particles is additionally altered after environmental exposure. We developed a microfluidic microscopy platform to demonstrate that the ζ-potential determines particle-cell adhesion strength. Furthermore, we find that due to this effect, the ζ-potential also strongly determines the internalization of the microplastic particles into cells. Therefore, the ζ-potential can act as a proxy of microplastic-cell interactions and may govern adverse effects reported in various organisms exposed to microplastics.

Detection and specific chemical identification of submillimeter plastic fragments in complex matrices such as compost.

Research on the plastic contamination of organic fertilizer (compost) has largely concentrated on particles and fragments > 1 mm. Small, submillimeter microplastic particles may be more hazardous to the environment. However, research on their presence in composts has been impeded by the difficulty to univocally identify small plastic particles in such complex matrices. Here a method is proposed for the analysis of particles between 0.01 and 1.0 mm according to number, size, and polymer type in compost. As a first demonstration of its potential, the method is used to determine large and small microplastic in composts from eight municipal compost producing plants: three simple biowaste composters, four plants processing greenery and cuttings and one two-stage biowaste digester-composter. While polyethylene, PE, tends to dominate among fragments > 1 mm, the microplastic fraction contained more polypropylene, PP. Whereas the contamination with PE/PP microplastic was similar over the investigated composts, only composts prepared from biowaste contained microplastic with a signature of biodegradable plastic, namely poly(butylene adipate co-terephthalate), PBAT. Moreover, in these composts PBAT microplastic tended to form the largest fraction. When the bulk of residual PBAT in the composts was analyzed by chloroform extraction, an inverse correlation between the number of particles > 0.01 mm and the total extracted amount was seen, arguing for breakdown into smaller particles, but not necessarily a mass reduction. PBAT oligomers and monomers as possible substrates for subsequent biodegradation were not found. Remaining microplastic will enter the environment with the composts, where its subsequent degradability depends on the local conditions and is to date largely uninvestigated.

Seasonal variations of microplastic pollution in the German River Weser.

Rivers play a major role in the distribution of microplastics (MPs) in the environment, however, research on temporal variations in these highly dynamic systems is still in its infancy. To date, most studies dealing with the seasonality of MP contamination in rivers focus on bi-yearly analysis, while temporal fluctuations over the course of the year are rarely studied. To shed more light on seasonal variability of MP abundance and potential driving factors, we have thus sampled the water surface of one location in the Weser River in Germany monthly over one entire year. In our study, we targeted MP in the size range 10-5000 µm, using two different state-of-the-art sampling methods (manta net for large MP (l-MP; 500-5000 µm) and a filtration system for small MP (s-MP; 10-500 µm)) and analysis techniques (ATR-FTIR and FPA-µFTIR) for chemical identification. Our findings show a strong size-dependent positive correlation of the MP concentration with discharge rates (specifically direct runoff) and suspended particulate matter (SPM) for s-MPs, specifically in the size range 10-149 µm. L-MPs, however, show a different environmental behaviour and do not follow these patterns. With our study, we were able to deliver a much higher temporal resolution, covering a broader size range of MPs compared to most studies. Our findings point towards an interplay of two possible mechanisms: a) the riverbeds play an important role in large-scale MP and SPM release via resuspension during high discharge events, and b) precipitation-driven soil erosion and runoff from urban surfaces (e.g. rain sewers) introduce MP and SPM. Hence, our study serves as a basis for more detailed investigations of MP transport in and between ecosystems.

The fate of microplastics from municipal wastewater in a surface flow treatment wetland.

Microplastics (MPs) are an anthropogenic pollutant of emerging concern prominent in both raw and treated municipal wastewater as well as urban and agricultural run-off. There is a critical need for the mitigation of both point- and diffuse sources, with treatment wetlands a possible sustainable nature-based solution. In this study, the possible retention of MPs in treatment wetlands of the widely used surface flow (SF) type was investigated. In- and outflow water, as well as atmospheric deposition, at a full-scale reed-based SF wetland (operating as a polishing phase of municipal wastewater treatment) was analyzed for MPs in a size range of 25-1000 µm. FPA-based µFT-IR spectroscopic imaging was used in combination with automated data analysis software, allowing for an unbiased assessment of MP numbers, polymer types and size distribution. Inflow water samples (secondary treated wastewater) contained 104 MPs m-3 and 56 MPs m-3 in sampling campaigns 1 and 2, respectively. Passage through the SF wetland increased the MP concentration in the water by 92 % during a rain intense period (campaign 1) and by 43 % during a low precipitation period (campaign 2). The MP particle numbers, size and polymer type distribution varied between the two sampling campaigns, making conclusions around the fate of specific types of MPs in SF wetlands difficult. Atmospheric deposition was measured to be 590 MPs m-2 week-1 during the rain-intense period. Our findings point towards atmospheric deposited MPs as an important factor in the fate of MPs in SF wetlands, causing an increase of MP concentrations, and potentially explaining the variations observed in MP concentrations in wetland effluent and removal efficiency. Furthermore, atmospheric deposition might also be a reason for the considerable inter-study variation regarding MPs removal efficiency in SF wetlands found in the available literature.

Effects of diesel exhaust particles on the health and survival of the buff-tailed bumblebee Bombus terrestris after acute and chronic oral exposure.

The diesel-powered transportation sector is a major producer of environmental pollution in the form of micro- and nanoscale diesel exhaust particles (DEP). Pollinators, such as wild bees, may inhale DEP or ingest it orally through plant nectar. However, if these insects are adversely affected by DEP is largely unknown. To investigate potential health threats of DEP to pollinators, we exposed individuals of Bombus terrestris to different concentrations of DEP. We analysed the polycyclic aromatic hydrocarbons (PAH) content of DEP since these are known to elicit adverse effects on invertebrates. We investigated the dose-dependent effects of those well-characterized DEP on survival and fat body content, as a proxy for the insects' health condition, in acute and chronic oral exposure experiments. Acute oral exposure to DEP showed no dose-dependent effects on survival or fat body content of B. terrestris. However, we could show dose-dependent effects after chronic oral exposure with high doses of DEP where significantly increased mortality was observed. Further, there was no dose-dependent effect of DEP on the fat body content after exposure. Our results give insights into how the accumulation of high concentrations of DEP e.g., near heavily trafficked sites, can influence insect pollinators' health and survival.

Role of Residual Monomers in the Manifestation of (Cyto)toxicity by Polystyrene Microplastic Model Particles.

Polystyrene (PS) is an important model polymer for the investigation of effects of microplastic (MP) and nanoplastic (NP) particles on living systems. Aqueous dispersions of PS MP or NP contain residual monomers of styrene. In consequence, it is not clear if the effects observed in standard (cyto)toxicity studies are evoked by the polymer (MP/NP) particle or by residual monomers. We addressed that question by comparing standard PS model particle dispersions with in-house synthesized PS particle dispersions. We proposed a rapid purification method of PS particle dispersions by dialysis against mixed solvents and developed a simple method of UV-vis spectrometry to detect residual styrene in the dispersions. We found that standard PS model particle dispersions, which contain residual monomers, exerted a low but significant cytotoxicity on mammalian cells, while the in-house synthesized PS, after rigorous purification to reduce the styrene content, did not. However, the PS particles per se but not the residual styrene in both PS particle dispersions resulted in immobilization of Daphnia. Only by using freshly monomer-depleted particles, will it be possible in the future to assess the (cyto)toxicities of PS particles, avoiding an otherwise not controllable bias effect of the monomer.

The complexity of micro- and nanoplastic research in the genus Daphnia - A systematic review of study variability and a meta-analysis of immobilization rates.

In recent years, the number of publications on nano- and microplastic particles (NMPs) effects on freshwater organisms has increased rapidly. Freshwater crustaceans of the genus Daphnia are widely used in ecotoxicological research as model organisms for assessing the impact of NMPs. However, the diversity of experimental designs in these studies makes conclusions about the general impact of NMPs on Daphnia challenging. To approach this, we systematically reviewed the literature on NMP effects on Daphnia and summarized the diversity of test organisms, experimental conditions, NMP properties and measured endpoints to identify gaps in our knowledge of NMP effects on Daphnia. We use a meta-analysis on mortality and immobilization rates extracted from the compiled literature to illustrate how NMP properties, study parameters and the biology of Daphnia can impact outcomes in toxicity bioassays. In addition, we investigate the extent to which the available data can be used to predict the toxicity of untested NMPs based on the extracted parameters. Based on our results, we argue that focusing on a more diverse set of NMP properties combined with a more detailed characterization of the particles in future studies will help to fill current research gaps, improve predictive models and allow the identification of NMP properties linked to toxicity.

Localisation and identification of polystyrene particles in tissue sections using Raman spectroscopic imaging.

The uptake of microplastic particles (MPP) by organisms is frequently described and poses a potential risk for these organisms and ultimately for humans either through direct uptake or trophic transfer. Currently, the in-situ detection of MPP in organisms is typically based on histological examination of tissue sections after uptake of fluorescently-labelled MPP and is thus not feasible for environmental samples. The alternative approach is purification of MPP from whole organisms or organs by chemical digestion and subsequent spectroscopic detection (FT-IR or Raman). While this approach is feasible for un-labelled particles it goes along with loss of any spatial information related to the location in the tissue. In our study we aimed at providing a workflow for the localisation and identification of non-fluorescent and fluorescent polystyrene (PS) particles (fragments, size range 2-130 µm) in tissue sections of the model organism Eisenia fetida with Raman spectroscopic imaging (RSI). We provide methodological approaches for the preparation of the samples, technical parameters for the RSI measurements and data analysis for PS differentiation in tissue sections. The developed approaches were combined in a workflow for the in-situ analysis of MPP in tissue sections. The spectroscopic analysis requires differentiation of spectra of MPP and interfering compounds, which is challenging given the complexity of tissue. Therefore, a classification algorithm was developed to differentiate PS particles from haem, intestinal contents and surrounding tissue. It allows the differentiation of PS particles from protein in the tissue of E. fetida with an accuracy of 95%. The smallest PS particle detected in the tissue was 2 µm in diameter. We show that it is possible to localise and identify non-fluorescent and fluorescent ingested PS particles directly in tissue sections of E. fetida in the gut lumen and the adjacent tissue.

Comparison of two rapid automated analysis tools for large FTIR microplastic datasets.

One of the biggest issues in microplastic (MP, plastic items <5 mm) research is the lack of standardisation and harmonisation in all fields, reaching from sampling methodology to sample purification, analytical methods and data analysis. This hampers comparability as well as reproducibility among studies. Concerning chemical analysis of MPs, Fourier-transform infrared (FTIR) spectroscocopy is one of the most powerful tools. Here, focal plane array (FPA) based micro-FTIR (µFTIR) imaging allows for rapid measurement and identification without manual preselection of putative MP and therefore enables large sample throughputs with high spatial resolution. The resulting huge datasets necessitate automated algorithms for data analysis in a reasonable time frame. Although solutions are available, little is known about the comparability or the level of reliability of their output. For the first time, within our study, we compare two well-established and frequently applied data analysis algorithms in regard to results in abundance, polymer composition and size distributions of MP (11-500 µm) derived from selected environmental water samples: (a) the siMPle analysis tool (systematic identification of MicroPlastics in the environment) in combination with MPAPP (MicroPlastic Automated Particle/fibre analysis Pipeline) and (b) the BPF (Bayreuth Particle Finder). The results of our comparison show an overall good accordance but also indicate discrepancies concerning certain polymer types/clusters as well as the smallest MP size classes. Our study further demonstrates that a detailed comparison of MP algorithms is an essential prerequisite for a better comparability of MP data.

Allelochemical run-off from the invasive terrestrial plant Impatiens glandulifera decreases defensibility in Daphnia.

Invasive species are a major threat for native ecosystems and organisms living within. They are reducing the biodiversity in invaded ecosystems, by outcompeting native species with e. g. novel substances. Invasive terrestrial plants can release allelochemicals, thereby reducing biodiversity due to the suppression of growth of native plants in invaded habitats. Aside from negative effects on plants, allelochemicals can affect other organisms such as mycorrhiza fungi and invertebrates in terrestrial ecosystems. When invasive plants grow in riparian zones, it is very likely that terrestrial borne allelochemicals can leach into the aquatic ecosystem. There, the often highly reactive compounds may not only elicit toxic effects to aquatic organisms, but they may also interfere with biotic interactions. Here we show that the allelochemical 2-methoxy-1,4-naphthoquinone (2-MNQ), produced by the ubiquitously occurring invasive terrestrial plant Impatiens glandulifera, interferes with the ability of Daphnia to defend itself against predators with morphological defences. Daphnia magna and Daphnia longicephala responded with morphological defences induced by chemical cues released by their corresponding predators, Triops cancriformis or Notonecta sp. However, predator cues in combination with 2-MNQ led to a reduction in the morphological defensive traits, body- and tail-spine length, in D. magna. In D. longicephala all tested inducible defensive traits were not significantly affected by 2-MNQ but indicate similar patterns, highlighting the importance to study different species to assess the risks for aquatic ecosystems. Since it is essential for Daphnia to adapt defences to the current predation risk, a maladaptation in defensive traits when simultaneously exposed to allelochemicals released by I. glandulifera, may therefore have knock-on effects on population dynamics across multiple trophic levels, as Daphnia is a key species in lentic ecosystems.

A novel approach to extract, purify, and fractionate microplastics from environmental matrices by isopycnic ultracentrifugation.

The increasing accumulation of microplastics (MP) in the environment is considered one of the most important environmental challenges of our times. Reliable extraction and detection methods for MP in environmental samples are essential for determining the extent of pollution and assessing ecological risks. However, extraction of MP from complex environmental matrices such as soil remains technically challenging. Today, density-based extractions with saturated salt solutions are widely applied. Nevertheless, current methods do not allow for the fractionation of different MP particle types according to their specific polymer densities. Here, we present a novel isopycnic ultracentrifugation approach for the simultaneous extraction and fractionation of MP mixtures based on the particle-specific buoyant densities. In this proof-of-concept study, diffusion-based density gradients were prepared using caesium chloride media, covering a density range between 1.1 and 1.5 g mL-1, sufficient to resolve many common polymer densities. We selected MP particles with a low (polyamide; PA66), medium (polybutylene adipate terephthalate; PBAT), and high (polyethylene terephthalate; PET) density to validate separation performance. Both pristine and soil-incubated MP mixtures showed clear banding patterns at expected buoyant densities after isopycnic separation. µFTIR imaging of subsamples collected from resolved MP fractions showed a polymer-specific separation of ≥87.6 %. In addition, the quantitative recovery of MP particles from soil was between 86 and 99 %. The potential of isopycnic ultracentrifugation to preserve MP-associated biofilms was also assessed. Soil-incubated MP particles were inspected by confocal laser scanning microscopy before and after isopycnic separation, indicating a preservation of bioorganic structures. Hence, isopycnic ultracentrifugation offers a powerful novel approach for a polymer-specific extraction and resolution of MP particles with a wide potential for applications in MP research.