Size distribution of nanoplastics and tyre wear particles in human tonsils

Size distribution of nanoplastics and tyre wear particles in human tonsils | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Size distribution of nanoplastics and tyre wear particles in human tonsils Dušan Materić, Vaishnavi Tokla, Tommy Cedervall, Rupert Holzinger, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6154338/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract This study investigates the presence of micro- and nanoplastics in human tonsil tissue. The tonsils are uniquely positioned in the oropharynx, a gateway to both the immune, respiratory, and digestive systems, thus acting as the first line of defence towards inhaled and ingested particles. We analysed tonsil samples from 15 individuals using Thermal Desorption - Proton Transfer Reaction - Mass Spectrometry, detecting a range of micro- and nanoplastics types, including polystyrene (PS), polyethylene (PE), and polyvinyl chloride (PVC), along with notable findings of tyre wear particles. We detected large differences in polymer types and size classes for each sample with concentrations spanning over four orders of magnitude, bringing nanoplastic concentration for the size class 20-200 nm with an average of 350 ng/mg dry weight. This study is the first to document the accumulation of nanoplastics and nanosized tyre wear in an immunologically active human organ. Biological sciences/Chemical biology Earth and environmental sciences/Environmental sciences/Environmental chemistry Physical sciences/Chemistry/Chemical biology Biological sciences/Biological techniques Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Plastic is considered the most widely used material in the world, as only in Europe the total plastics production reached 54 million tons in 2023 1,2 . The worldwide production of plastic has grown exponentially in just a few decades – from 2 Mt in 1950 to 413.8 Mt in 2023 – and with it, the amount of plastic waste 2 . Historically, plastics have attracted considerable attention due to their low production costand favourable characteristics, including light weight, durability, flexibility, and water resistance. However, the same attributes that make plastics desirable—especially durability and resistance to degradation—also naturally contribute to their long-term persistence in the environment. Thus, plastics are not easily biodegradable and can survive for centuries 3 , and their pollution is recognised to be one of the greatest environmental challenges of the 21st century: they might cause wide-ranging damage to ecosystems 4 and human health 5–7 . Plastics are made from synthetic or semisynthetic polymers, which consist of long, repeating chains. Various additives, such as plasticisers, flame retardants, and colourants, are often incorporated to improve their properties and appearance 8–10 . In Europe, the most widely used plastics are polypropylene (PP) and polyethylene (PE), including its low-density form (LDPE), followed by polyvinyl chloride (PVC), polyethylene terephthalate (PET), and polystyrene (PS) 2 . The growing environmental consequences of plastic pollution have been topics of considerable interest in recent years, especially small plastic particles known as microplastics and nanoplastics. Microplastics are generally defined as plastic particles that are less than 5 mm in size, whereas nanoplastics are even smaller, these are often defined as plastic particles that are less than 1 µm in size 11,12 . These plastics are mostly formed from the breakdown of larger plastic items in the environment; however, many cosmetics and skincare products contain intentionally produced and added microbeads, which, to a lesser extent, contribute to overall environmental microplastic pollution 13 . Micro- and nanoplastics are ubiquitous. Studies have found microplastic particles in various environmental compartments. For instance, microplastics have been found in river sediments 14,15 , agricultural soil 16 , oceans 17–20 and even in remote regions such as the deep sea 21,22 or the Antarctic ice 23–25 . The detection and characterisation of nanoplastics presents technical challenges 26,27 . However, there is a limited number of studies indicating that nanoplastics can be found wherever microplastics are detected 28 . Correspondingly, nanoplastics have been found, for example, in the Dutch Wadden sea 20 , southern and northern polar ice 29 , and in alpine snow and glaciers 30,31 , atmospheric fallouts 32 and urban 33 , rural and remote air 34 . The ubiquitous occurrence of micro- and nanoplastics, even in remote regions such as the deep sea or Antarctica, shows that these particles are transported globally. In addition to the inanimate environmental samples, microplastic particles have also been found in a variety of organisms and human tissues. In humans, microplastics have been detected in blood, 35 liver, 36 lung, 37 kidney and brain tissue 38 as well as throughout the reproductive system, including semen, 39 testes, 40,41 uterus, 42 and placenta. 43 While their effects have not yet been fully characterised, exposure to plastics has been linked to cell apoptosis and both genotoxicity and cytotoxicity. 44–46 Additionally, plastic particles may release plasticisers/additives and adsorbed pollutants, potentially enhancing their effects 9,10 . The prevalence of small plastic particles in human beings is somewhat known, but no study has shown nanosized particles originating from tyres in human organs. Nanoplastics and tyre wear particles have the potential to cause significant impact due to their small size and large surface area. However, the effects of these widely dispersed nanoparticles on human health have yet to be thoroughly investigated. In humans, the tonsils are the first immunologically active tissue that encounters particles upon oral and nasal exposure. Tonsils are located in the oral cavity, and air, food and liquid pass the tonsils on route to the respiratory and digestive tracts. Their position optimises the uptake of foreign materials like antigens and possibly micro- and nanoplastics. In this work, we aimed to quantify presence and size distribution and types of micro- and nanoplastics in tonsil tissues and hypothesised that tonsils, besides skin, digestive and respiratory systems, could play an active and significant role in human micro- and nanoplastics exposure pathway. Material & Methods Tonsil samples The tonsils were obtained from individuals who underwent conventional tonsillectomy at the Otorhinolaryngology department in Ystad, southern Sweden. The indications for tonsillectomy are sleep apnoea or chronic tonsilitis. The tonsils would, if not used in the study, have been destroyed. Prior to sampling, all patients signed informed consent. The consent form was signed by the legal guardians of underage patients (<18 years old). Following surgery, the tonsils were placed in 50 mL falcon tubes (Eppendorf, Sweden) and stored one night in a freezer and then delivered to the Department of Otorhinolaryngology in Lund, southern Sweden, and then further on to the Department of Biochemistry and Structural Biology at Lund University, where they were stored at -20 °C until further analysis began. The study was approved by The Regional Ethical Committee (Dnr 2018/611) and the Regional Bioethical Committee (136/BD16). There were 15 tonsils included in the study from 15 different patients with a mean age of 18 years (min-max 3-44 years), of which 9 were female and 8 were children (20 cigarettes per day. Ten of the patients were living in a city with more than 100000 inhabitants. It was common to drink and eat from plastic containers; 10 persons were drinking from a plastic bottle more than twice a week, and six had a daily drink from a bottle made of plastic. Eating from plastic containers daily was reported by 6, and at least twice a week by 12 persons. This data is summarised in Table S2. Chemicals and materials Potassium hydroxide (KOH) pellets were obtained from Sigma-Aldrich, and a hydrogen peroxide (H 2 O 2 , 3%) solution was supplied by Frost Pharma. All the solutions were prepared using HPLC water (VWR, Germany), and the digestion solution was freshly prepared daily. Glass fibre filters were purchased from Whatman TM , while quartz fibre filters, anodisc filters, and the glass filtration setup were supplied by Sterlitech. The isolation of plastic particles from the tonsil tissue was evaluated using polystyrene particles of 0.5 μm size, sourced from Sigma-Aldrich. These particles were used to spike samples for recovery experiments. The same chemicals and materials are used for the samples and complete procedural blanks. Sample preparation The tonsil tissues were removed from the freezer and rinsed four times with Milli-Q water to remove any residual blood from the surgery. The tissues were then freeze-dried for 24 hours at −55°C, grounded using a ceramic mortar, and weighed. To prevent contamination and rehydration, the tonsil tissues were stored in a desiccator and wrapped in aluminium foil. The digestion and extraction of nanoplastics were performed with slight modifications to the previously published protocol 48,57 . Tonsil tissue samples of a known quantity of 10 mg were transferred into glass bottles. To each bottle, 8 mL of digestion solution consisting of 2.5% KOH and 3% H 2 O 2 in a 1:1 ratio was added. The bottles were then incubated in a water bath at 60°C for 8 hours to facilitate the breakdown of organic matter. After incubation, the samples were allowed to cool to room temperature, and the solutions were neutralised by the dropwise addition of 10% citric acid until a neutral pH was achieved. Solutions were filtered in cascade using different filter membranes and pore sizes including 2700 nm (Glass fibre filter), 1200 nm (Silver membrane filter), 200 nm (Anodisc filter), and 20 nm (Anodisc filter). Each bottle was rinsed twice with HPLC water. After each filtration step, the filters were washed with HPLC water to ensure no residual particles remained. Each filter was dried and stored in clean, sterile glass petri dishes for further analysis. Additionally, two procedural blanks were included to evaluate potential contamination and impurities arising from laboratory equipment and the procedural methodology. Following the filtration, the filters were dried and cut in a laminar flow workstation. For the glass fibre and silver membrane filters, cutting was performed using a metal punch tool with a 3 mm diameter. The anodisc filters (200 nm and 20 nm pore size) were cracked using a spatula and tweezers. The resulting filter pieces were photographed, and their surface areas were estimated using ImageJ software 58 . Tools and surfaces were cleaned with ethanol and cellulose tissue between uses. Three aliquots from each filter stage of each sample and blank were then transferred into pre-baked (250°C, overnight) 10 mL glass vials. TD-PTR-MS Analysis The samples were analysed using thermal desorption proton transfer mass spectrometry (TD-PTR-MS), as previously described in detail 47,59 . Briefly, the TD-PTR-MS protocol was carried with the following parameters: a temperature ramp from 35°C to 360°C, an E/N ratio of 120 Td, and a p-drift pressure of 2.9 mbar to minimise clustering and fragmentation. TD-PTR-MS analysis was performed using a PTR8000 instrument (Ionicon Analitik, AU). Both samples and blanks were analysed in triplicates to ensure data reliability and reproducibility. The first level of data processing (peak identification, integration, TOF calibration, ion quantification) was conducted using the PTRwid software 60 . The TD-PTR-MS signal was integrated over a 7-minute period, starting when the thermal desorption temperature reached 200°C, to capture the thermal degradation products of each polymer. The mass spectra were corrected by subtracting the blank signal, and organic ions below the 3σ detection limit were excluded from further analysis. TD-PTR-MS has been applied for the analysis of organic aerosols 61 , dissolved organic matter (DOM) 62 , Methylsiloxanes 63,64 , and nanoplastics in different environments, including seawater 20 , surface water 65 , snow 47 , ice 29 , glaciers 31 , atmospheric samples 33,34 , and animal tissue 48 . Similarly, here, we obtained plastic identification via the fingerprinting approach, which utilises the 40 most abundant library ions with m/z >100 for the identification and quantification of microplastics and nanoplastics in the samples. Fingerprinting was performed on all samples and blanks (see Data Availability section for all the steps of the analysis). In short, the ion signals of each sample were compared to those in the reference library to identify matching patterns (Figure 4). The similarity was assessed based on the ratio of ion signals from the library using four different matching algorithms. If a match was obtained (z-score > 2), each identified ion present in the plastics—after contamination correction—was quantified following the standard PTR-MS protocol 49,50 . Details on data processing and matching algorithms can be found in previous studies, e.g. 20,47,48 , and the standard PTR-MS quantification method is explained elsewhere 49–51,66 . A step-by-step outline of our data analysis and processing, from raw files to final results, is provided in the supplementary information and Data Availability section. Quality Control and Assurance (QC/QA) Given that the presence of plastics is unavoidable, extra care was taken to prevent contamination of the samples. A laminar flow was used to avoid contamination from the surroundings. Laboratory staff wore cotton lab coats and nitrile gloves. Additional precautions were taken to minimise contamination, such as using silicone and Teflon materials, with silicone vacuum tubing and Teflon discs replaced on the glass vial seals. All glassware was heat-treated at 250°C overnight, and all containers were covered with aluminium foil to prevent airborne plastic contamination. Procedural blanks were included to assess potential contamination from the laboratory environment, reagents, or equipment. Our experiments included three different blanks: a) Procedural blanks to evaluate contamination during laboratory operations. These represent control samples treated the same way as the actual tonsil samples but without tonsil tissue, allowing for the identification of any plastics introduced by the laboratory environment or procedures. b) Filter blanks to measure possible contamination from the filter material. c) System blanks to assess contamination from the instrument. In the procedural blanks, some traces of plastic were detected. For example, extraction batch that included patient 7 and associated procedural blanks, we detected an average of 3.7 ng absolute of tyre particles in the procedural blanks, while the sample itself contained 37–707 ng absolute (see Supplementary Information and Data Availability for details). Furthermore, the results showed that the plastic content in the samples was significantly higher than in the procedural blanks for all plastic types, indicating that most of the plastics detected—such as tyre particles, PET, PS, PVC, PP/PPC, and PE—originated from the samples rather than from contamination during the experimental process. This confirms the presence of various plastics in the actual samples, particularly PVC and PP/PPC, which were absent in the blanks for nanoplastics. All reported values were corrected using the mean blank value for each extraction batch. The tonsils samples were collected specifically for the project and transport from surgery theatre has been planned to reduce contamination with short way from patient into sample storing container. Potential contamination is still possible and it was experimentally reduced by washing the organs and avoiding sampling from the tissue surface, which could have been exposed to external contaminants. However, we acknowledge that the best blank control would have been archived tonsil samples from the pre-plastic era, but we did not have access to such specimens. Additionally, to evaluate the ionisation efficiency and recovery rate of our analytical method, we performed spiking assessments on both procedural blanks and sample filters by adding a known quantity of PS particles to random runs. The ionisation efficiency and recovery rate averaged 30%, which is similar to previously obtained values 31,48 . Furthermore, spiking the samples with PS showed good match scores (z-score > 4 for at least one algorithm), suggesting that no matrix interference (e.g., from insufficient tissue digestion) compromised the plastic fingerprinting process. Ideally, further analysis of such samples, besides TD-PTR-MS, could be coupled with optical methods that facilitate the chemical identification of polymers and have the same or a similar analytical window, such as Raman spectroscopy. TD-PTR-MS and Raman have previously been used in tandem, yielding overall similar results 32 ; however, this was achieved for relatively clear matrices, such as atmospheric deposition. In this project, which involved human samples, such cross-comparison of different methods was not technically possible. Developing a digestion protocol and a pre-concentration method compatible with both Raman spectroscopy and TD-PTR-MS should be a future focus of analytical development to improve QC/QA. Assessing the uncertainties In this study, we performed replicated measurements to assess the experimental uncertainty in our analysis of micro- and nanoplastics. We also included procedural blanks to apply suitable corrections for any plastics that might have been introduced during sample handling. However, additional factors could contribute to both underestimation and overestimation. Here, we outline the potential causes and assess the associated uncertainties. Underestimation The most prominent factor leading to underestimation is related to ionisation efficiency and recovery in proton transfer reaction mass spectrometry. We tested ionisation efficiency and recovery by spiking blanks with a known amount of an analytical-grade PS standard, obtaining an average recovery of 29.7% (see SI). However, quantitative analytical standards for other nanoplastics do not exist, and we cautiously assume that this value holds, on average, for other plastic types. Additionally, we account for the PTR-MS uncertainty of ±30% 50 . Using these two parameters, we calculate the underestimation factor as: This suggests that our reported concentrations may be underestimated by a factor of 3.7. Overestimation Overestimation may occur if organic matter interferes with the ions used for identification and quantification. Such interference can arise, for example, from incomplete digestion of organic tissue. To estimate this effect, we performed a Monte Carlo simulation 67 to assess the impact of organic matter (OM) contamination on our 40 marker ions (see SI). In this simulation, we stepwise increased OM signal contributions by up to 350%, repeating each step 1000 times. The results, summarised in Figure 5, indicate that increased organic matter can lead to overestimation, but only up to 31%. Beyond this threshold, the 40-ion plastic fingerprinting approach prevents further overestimation. For example, when organic matter impurities increase by 200% over the 40 PET-associated ions, overestimation rises to 31%, but further increases in OM distort the mass spectra, preventing a positive PET match. This Monte Carlo analysis highlights the importance of a multi-ion approach, which is significantly more resistant to false positives than traditional methods relying on one or a few ions, such as py-GC-MS or TED-GC-MS. Using the Monte Carlo values and a PTR-MS uncertainty of 30%, we calculate total overestimation uncertainty as: 0.31 + (0.31×30%) = 0.40 This suggests that our overall overestimation could be up to 40%. Results & Discussions This study shows the first data of micro- and nanoplastics as well as tyre size distribution in human tonsils. Our conservative approach to nanoplastic measurements relies on strict quality control of the analysis, including tested digestion protocol, multiple blanks (including complete procedural blanks), triplicate samples, spiking experiments with known amount of nanoplastics, conservative plastic detection using 40-ion fingerprinting and detection 47,48 for each plastic type and plastics quantification based on well-established PTR-TOF-MS quantification procedures 49–51 widely applied in atmospheric, environmental, and food analysis. Details on QC/QA including detailed assessment of the uncertainties can be found in the Methods section. All 15 samples from different individuals showed the presence of at least one type of micro- and/or nanoplastics, spanning the mass concentration over four orders of magnitude (from 1 to 12000 ng/mg). The mean nanoplastics concentration for the 20-200 nm particle size was 350 ng/mg DW (median 126 ng/mg DW), reaching over 1200 ng/mg DW. Microplastic loads (size >2700 nm) were, on average, 2500 ng/mg (median 2083 ng/mg DW) and reached 9400 ng/g DW of tonsil tissue (Table S1). The average size distribution for all plastic types analysed in the samples is shown in Figure 1. For most of the samples (n=14), we measured the mass increase either on the 2700 nm filter or cut-off size fraction 20-200 nm or both. Comparatively, little plastic mass was observed in the size range of 200 – 1200 nm, and 1200 -2700 nm. The primary polymer types for microplastics were tyre wear, PVC, and PS, while for nanoplastics, the major types were tyre wear, PE, and PVC (Figure 1b and c). To our knowledge, it is the first time tyre wear nanoparticles have been detected in human tissues. However, the presence of most polymer types was evident in all size ranges, but it was somewhat different for each sample (Figure 2). We did not observe the absolute dominance of the PE polymer as reported in previous studies for microplastics in human samples (including placenta, artery plaque, kidney, liver and brain) 38,52,53 . In fact, in 9 out of 15 samples (60%), we did not detect any PE nanoplastics, and in 10 out of 15 samples (different once than for PE), we did not detect PVC nanoplastics either. This discrepancy can be explained by methodological differences, as recent findings suggest that PE and PVC could be falsely assigned as present in py-GC-MS. In contrast, TD-PTR-MS, with its high resolution and 40-ion identification approach, provides much more conservative data for nanoplastics in complex matrices compared to py-GC-MS. We observe a considerable variation in plastic loads and types across different size fractions and patients (Figure 2). This significant variation in the load of specific plastic types and sizes is likely due to vast differences in individual exposure. Notably, some polymer types are absent in certain patients, while others appear only within specific size groups. This suggests that exposure to different plastic types may be influenced by particle size—e.g., depending on the initial size distribution of plastics in air or food. These findings highlight the need for future systematic and comprehensive analysis of micro- and nanoplastic size distribution in indoor and occupational environments, as well as in food and beverages that humans are exposed to. To our knowledge, the quantity of nanoplastics and size distribution of the plastic for biological samples is only reported for seashells – mussels 48 , and a similar analytical approach is used. The human tonsils contained 4 times more nanoplastics and 20 times more microplastics than the farmed mussels. Both these studies included a limited number of samples and showed a vast diversity of NPs concentrations. Sizes and types of micro- and nanoplastics vary from tonsil to tonsil in terms of concentration and plastic type, as exemplified in Figure 3. This additionally shows that human exposure might be highly diverse on a case-to-case basis. Using the same method, we previously measured tyre wear particles in farmed mussels, which contributed around 6% of total nanoplastics and only 1% of microplastics load, respectively 48 . However, for the human tonsil tissue, we observed a relative tyre wear contribution of 27% for nanoplastics and 32% for microplastics. On average, in tonsils, the mass concentrations were 6 times higher for microplastics (>2700 nm) compared to the nanoplastics (20-200 nm). However, particle number concentrations, considering the most conservative sizes for micro- and nanoplastics (2700 and 200 nm respectively), and average concentrations (2500 ng/mg for MPs and 350 ng/mg or NPs), estimate the average particle number of microplastics to ~127013 particles and nanoplastics to ~43750000 particles. In other words, our data shows that, on average, there are 300 times more nanoplastics than microplastic particles in the tonsil samples. This also results in a larger surface area, which can be important for the effects on human health as many reactions occur on surfaces dependent on size and material. For example, the effect on blood coagulation from polystyrene nanoparticles depends on particle surface chemistry 54 and on particle size 55 . However, nanoplastic mechanisms and kinetics around primary uptake, possibly passing into the lymph system or bloodstream, and excretion mechanisms are still mostly theoretical. Notably, the larger quantities of higher microplastic size fractions (Figures 1a and 2) compared to nanoplastics are consistent with the role of the tonsils as organs that actively sample fine particles for immune response. However, this does not mean that such large plastic particles can effectively enter the lymphatic system and the blood. It is clear that more research is needed to understand the processes that may involve the activation of plastic particles of different sizes and polymer types, as well as their transport kinetics within and between organs in the human body. Given the limited number of samples, it is not possible to draw conclusions about plastic concentrations at the population level, especially considering the large variation described above. However, it is interesting to note that we observed a weak correlation between body mass index (BMI) and the concentration of PE nanoplastics (size fraction: 20–200 nm; R = 0.69, p = 0.0047, see SI). Previous studies have suggested that fatty acids and lipid structures may facilitate the retention of plastic particles (PS and PMMA—polymethyl methacrylate) 56 , but this has not yet been studied in vivo. The relationship between nanoplastic loads and BMI, as well as the mechanisms of retention in fat-rich tissues, would be an important topic for future research with potentially significant toxicological implications. Declarations Acknowledgements We want to thank Bengt Olsson, Anna Elfvik, and Natalia Ioukhnenko, and the doctors and the department of ORL surgery, Department of ORL, Ystad Hospital, Ystad, Sweden, for tonsil sample collection, Lena Glantz-Larsson, Department of ORL, Head and Neck Surgery, Skåne University Hospital, Lund, Sweden, for logistics support, and Morgan Andersson (deceased), Department of Otorhinolaryngology (ORL), Head and Neck Surgery, Skåne University Hospital, Lund, Sweden, for being part of study start-up. Author contributions. M.V., T.C., M.T.E., and DM conceived the project and designed the study. M.V. organised the sample collection and coordinated the project. D.M designed and coordinated the analysis and data collection. R.H provided the instrumentation. V.R.T. performed experiments supervised by M.T.E. and DM. V.R.T and DM performed data analysis and figure preparation. M.V, T.C., and DM provided funding support and/or the resources. DM, V.R.T, M.V. and wrote the original draft. All authors contributed to the manuscript editing. Data Availability: All data needed to evaluate the conclusions in the paper including raw files (measures related to nanoplastics) and processing stages of all the data analysis are available via: DOI:[the link will be available upon the review of the manuscript] Competing interests. No competing interests References Jambeck, J. R.; Walker-Franklin, I. The Impacts of Plastics’ Life Cycle. One Earth 2023 , 6 (6), 600–606. https://doi.org/10.1016/j.oneear.2023.05.015. Plastics Europe. Plastics - the Facts 2022 . https://plasticseurope.org/knowledge-hub/plastics-the-facts-2022/ (accessed 2023-09-04). Andrady, A. L.; Neal, M. A. Applications and Societal Benefits of Plastics. Philosophical Transactions of the Royal Society B: Biological Sciences 2009 , 364 (1526), 1977–1984. https://doi.org/10.1098/rstb.2008.0304. Ekvall, M. T.; Stábile, F.; Hansson, L.-A. Nanoplastics Rewire Freshwater Food Webs. Commun Earth Environ 2024 , 5 (1), 1–7. https://doi.org/10.1038/s43247-024-01646-7. Wright, S. L.; Kelly, F. J. Plastic and Human Health: A Micro Issue? Environ. Sci. Technol. 2017 , 51 (12), 6634–6647. https://doi.org/10.1021/acs.est.7b00423. Revel, M.; Châtel, A.; Mouneyrac, C. Micro(Nano)Plastics: A Threat to Human Health? Current Opinion in Environmental Science & Health 2018 , 1 , 17–23. https://doi.org/10.1016/j.coesh.2017.10.003. Ramsperger, A. F. R. M.; Bergamaschi, E.; Panizzolo, M.; Fenoglio, I.; Barbero, F.; Peters, R.; Undas, A.; Purker, S.; Giese, B.; Lalyer, C. R.; Tamargo, A.; Moreno-Arribas, M. V.; Grossart, H.-P.; Kühnel, D.; Dietrich, J.; Paulsen, F.; Afanou, A. K.; Zienolddiny-Narui, S.; Eriksen Hammer, S.; Kringlen Ervik, T.; Graff, P.; Brinchmann, B. C.; Nordby, K.-C.; Wallin, H.; Nassi, M.; Benetti, F.; Zanella, M.; Brehm, J.; Kress, H.; Löder, M. G. J.; Laforsch, C. Nano- and Microplastics: A Comprehensive Review on Their Exposure Routes, Translocation, and Fate in Humans. NanoImpact 2023 , 29 , 100441. https://doi.org/10.1016/j.impact.2022.100441. Bachmann, M.; Zibunas, C.; Hartmann, J.; Tulus, V.; Suh, S.; Guillén-Gosálbez, G.; Bardow, A. Towards Circular Plastics within Planetary Boundaries. Nat Sustain 2023 , 6 (5), 599–610. https://doi.org/10.1038/s41893-022-01054-9. Hahladakis, J. N.; Velis, C. A.; Weber, R.; Iacovidou, E.; Purnell, P. An Overview of Chemical Additives Present in Plastics: Migration, Release, Fate and Environmental Impact during Their Use, Disposal and Recycling. J Hazard Mater 2018 , 344 , 179–199. https://doi.org/10.1016/j.jhazmat.2017.10.014. Wiesinger, H.; Wang, Z.; Hellweg, S. Deep Dive into Plastic Monomers, Additives, and Processing Aids. Environ. Sci. Technol. 2021 , 55 (13), 9339–9351. https://doi.org/10.1021/acs.est.1c00976. Gigault, J.; El Hadri, H.; Nguyen, B.; Grassl, B.; Rowenczyk, L.; Tufenkji, N.; Feng, S.; Wiesner, M. Nanoplastics Are Neither Microplastics nor Engineered Nanoparticles. Nat. Nanotechnol. 2021 , 16 (5), 501–507. https://doi.org/10.1038/s41565-021-00886-4. Frias, J. P. G. L.; Nash, R. Microplastics: Finding a Consensus on the Definition. Marine Pollution Bulletin 2019 , 138 , 145–147. https://doi.org/10.1016/j.marpolbul.2018.11.022. Vethaak, A. D.; Legler, J. Microplastics and Human Health. Science 2021 , 371 (6530), 672–674. https://doi.org/10.1126/science.abe5041. Castañeda, R. A.; Avlijas, S.; Simard, M. A.; Ricciardi, A. Microplastic Pollution in St. Lawrence River Sediments. Can. J. Fish. Aquat. Sci. 2014 , 71 (12), 1767–1771. https://doi.org/10.1139/cjfas-2014-0281. Dekiff, J. H.; Remy, D.; Klasmeier, J.; Fries, E. Occurrence and Spatial Distribution of Microplastics in Sediments from Norderney. Environmental Pollution 2014 , 186 , 248–256. https://doi.org/10.1016/j.envpol.2013.11.019. Kumar, M.; Xiong, X.; He, M.; Tsang, D. C. W.; Gupta, J.; Khan, E.; Harrad, S.; Hou, D.; Ok, Y. S.; Bolan, N. S. Microplastics as Pollutants in Agricultural Soils. Environmental Pollution 2020 , 265 , 114980. https://doi.org/10.1016/j.envpol.2020.114980. Isobe, A.; Uchiyama-Matsumoto, K.; Uchida, K.; Tokai, T. Microplastics in the Southern Ocean. Marine Pollution Bulletin 2017 , 114 (1), 623–626. https://doi.org/10.1016/j.marpolbul.2016.09.037. Galgani, F.; Brien, A. S.; Weis, J.; Ioakeimidis, C.; Schuyler, Q.; Makarenko, I.; Griffiths, H.; Bondareff, J.; Vethaak, D.; Deidun, A.; Sobral, P.; Topouzelis, K.; Vlahos, P.; Lana, F.; Hassellov, M.; Gerigny, O.; Arsonina, B.; Ambulkar, A.; Azzaro, M.; Bebianno, M. J. Are Litter, Plastic and Microplastic Quantities Increasing in the Ocean? Microplastics and Nanoplastics 2021 , 1 (1), 2. https://doi.org/10.1186/s43591-020-00002-8. Haward, M. Plastic Pollution of the World’s Seas and Oceans as a Contemporary Challenge in Ocean Governance. Nat Commun 2018 , 9 (1), 667. https://doi.org/10.1038/s41467-018-03104-3. Materić, D.; Holzinger, R.; Niemann, H. Nanoplastics and Ultrafine Microplastic in the Dutch Wadden Sea – The Hidden Plastics Debris? Science of The Total Environment 2022 , 846 , 157371. https://doi.org/10.1016/j.scitotenv.2022.157371. Zhu, X.; Rochman, C. M.; Hardesty, B. D.; Wilcox, C. Plastics in the Deep Sea – A Global Estimate of the Ocean Floor Reservoir. Deep Sea Research Part I: Oceanographic Research Papers 2024 , 206 , 104266. https://doi.org/10.1016/j.dsr.2024.104266. Van Cauwenberghe, L.; Vanreusel, A.; Mees, J.; Janssen, C. R. Microplastic Pollution in Deep-Sea Sediments. Environmental Pollution 2013 , 182 , 495–499. https://doi.org/10.1016/j.envpol.2013.08.013. Finger, J. V. G.; Corá, D. H.; Convey, P.; Cruz, F. S.; Petry, M. V.; Krüger, L. Anthropogenic Debris in an Antarctic Specially Protected Area in the Maritime Antarctic. Marine Pollution Bulletin 2021 , 172 , 112921. https://doi.org/10.1016/j.marpolbul.2021.112921. Mishra, A. K.; Singh, J.; Mishra, P. P. Microplastics in Polar Regions: An Early Warning to the World’s Pristine Ecosystem. Science of The Total Environment 2021 , 784 , 147149. https://doi.org/10.1016/j.scitotenv.2021.147149. Kelly, A.; Lannuzel, D.; Rodemann, T.; Meiners, K. M.; Auman, H. J. Microplastic Contamination in East Antarctic Sea Ice. Marine Pollution Bulletin 2020 , 154 , 111130. https://doi.org/10.1016/j.marpolbul.2020.111130. Velimirovic, M.; Tirez, K.; Voorspoels, S.; Vanhaecke, F. Recent Developments in Mass Spectrometry for the Characterization of Micro- and Nanoscale Plastic Debris in the Environment. Anal Bioanal Chem 2020 . https://doi.org/10.1007/s00216-020-02898-w. Ivleva, N. P. Chemical Analysis of Microplastics and Nanoplastics: Challenges, Advanced Methods, and Perspectives. Chem. Rev. 2021 , 121 (19), 11886–11936. https://doi.org/10.1021/acs.chemrev.1c00178. Jakubowicz, I.; Enebro, J.; Yarahmadi, N. Challenges in the Search for Nanoplastics in the Environment—A Critical Review from the Polymer Science Perspective. Polymer Testing 2021 , 93 , 106953. https://doi.org/10.1016/j.polymertesting.2020.106953. Materić, D.; Kjær, H. A.; Vallelonga, P.; Tison, J.-L.; Röckmann, T.; Holzinger, R. Nanoplastics Measurements in Northern and Southern Polar Ice. Environmental Research 2022 , 208 , 112741. https://doi.org/10.1016/j.envres.2022.112741. Materić, D.; Ludewig, E.; Brunner, D.; Röckmann, T.; Holzinger, R. Nanoplastics Transport to the Remote, High-Altitude Alps. Environmental Pollution 2021 , 117697. https://doi.org/10.1016/j.envpol.2021.117697. Jurkschat, L.; Gill, A. J.; Milner, R.; Holzinger, R.; Evangeliou, N.; Eckhardt, S.; Materić, D. Using a Citizen Science Approach to Assess Nanoplastics Pollution in Remote High-Altitude Glaciers. Sci Rep 2025 , 15 (1), 1864. https://doi.org/10.1038/s41598-024-84210-9. Allen, S.; Materić, D.; Allen, D.; MacDonald, A.; Holzinger, R.; Roux, G. L.; Phoenix, V. R. An Early Comparison of Nano to Microplastic Mass in a Remote Catchment’s Atmospheric Deposition. Journal of Hazardous Materials Advances 2022 , 7 , 100104. https://doi.org/10.1016/j.hazadv.2022.100104. Kirchsteiger, B.; Materić, D.; Happenhofer, F.; Holzinger, R.; Kasper-Giebl, A. Fine Micro- and Nanoplastics Particles (PM2.5) in Urban Air and Their Relation to Polycyclic Aromatic Hydrocarbons. Atmospheric Environment 2023 , 301 , 119670. https://doi.org/10.1016/j.atmosenv.2023.119670. Kau, D.; Materić, D.; Holzinger, R.; Kasper-Giebl, A. Fine Microplastics and Nanoplastics in Particulate Matter Samples from a High Alpine Environment ; EGU23-5730; Copernicus Meetings, 2023. https://doi.org/10.5194/egusphere-egu23-5730. V. L. Leonard, S.; Liddle, C. R.; Atherall, C. A.; Chapman, E.; Watkins, M.; D. J. Calaminus, S.; Rotchell, J. M. Microplastics in Human Blood: Polymer Types, Concentrations and Characterisation Using μFTIR. Environment International 2024 , 188 , 108751. https://doi.org/10.1016/j.envint.2024.108751. Horvatits, T.; Tamminga, M.; Liu, B.; Sebode, M.; Carambia, A.; Fischer, L.; Püschel, K.; Huber, S.; Fischer, E. K. Microplastics Detected in Cirrhotic Liver Tissue. eBioMedicine 2022 , 82 . https://doi.org/10.1016/j.ebiom.2022.104147. Jenner, L. C.; Rotchell, J. M.; Bennett, R. T.; Cowen, M.; Tentzeris, V.; Sadofsky, L. R. Detection of Microplastics in Human Lung Tissue Using μFTIR Spectroscopy. Science of The Total Environment 2022 , 831 , 154907. https://doi.org/10.1016/j.scitotenv.2022.154907. Nihart, A. J.; Garcia, M. A.; El Hayek, E.; Liu, R.; Olewine, M.; Kingston, J. D.; Castillo, E. F.; Gullapalli, R. R.; Howard, T.; Bleske, B.; Scott, J.; Gonzalez-Estrella, J.; Gross, J. M.; Spilde, M.; Adolphi, N. L.; Gallego, D. F.; Jarrell, H. S.; Dvorscak, G.; Zuluaga-Ruiz, M. E.; West, A. B.; Campen, M. J. Bioaccumulation of Microplastics in Decedent Human Brains. Nat Med 2025 , 1–6. https://doi.org/10.1038/s41591-024-03453-1. Montano, L.; Giorgini, E.; Notarstefano, V.; Notari, T.; Ricciardi, M.; Piscopo, M.; Motta, O. Raman Microspectroscopy Evidence of Microplastics in Human Semen. Science of The Total Environment 2023 , 901 , 165922. https://doi.org/10.1016/j.scitotenv.2023.165922. Zhao, Q.; Zhu, L.; Weng, J.; Jin, Z.; Cao, Y.; Jiang, H.; Zhang, Z. Detection and Characterization of Microplastics in the Human Testis and Semen. Science of The Total Environment 2023 , 877 , 162713. https://doi.org/10.1016/j.scitotenv.2023.162713. Hu, C. J.; Garcia, M. A.; Nihart, A.; Liu, R.; Yin, L.; Adolphi, N.; Gallego, D. F.; Kang, H.; Campen, M. J.; Yu, X. Microplastic Presence in Dog and Human Testis and Its Potential Association with Sperm Count and Weights of Testis and Epididymis. Toxicological Sciences 2024 , 200 (2), 235–240. https://doi.org/10.1093/toxsci/kfae060. Xu, H.; Dong, C.; Yu, Z.; Hu, Z.; Yu, J.; Ma, D.; Yao, W.; Qi, X.; Ozaki, Y.; Xie, Y. First Identification of Microplastics in Human Uterine Fibroids and Myometrium. Environmental Pollution 2024 , 360 , 124632. https://doi.org/10.1016/j.envpol.2024.124632. Ragusa, A.; Svelato, A.; Santacroce, C.; Catalano, P.; Notarstefano, V.; Carnevali, O.; Papa, F.; Rongioletti, M. C. A.; Baiocco, F.; Draghi, S.; D’Amore, E.; Rinaldo, D.; Matta, M.; Giorgini, E. Plasticenta: First Evidence of Microplastics in Human Placenta. Environment International 2021 , 146 , 106274. https://doi.org/10.1016/j.envint.2020.106274. Lehner, R.; Weder, C.; Petri-Fink, A.; Rothen-Rutishauser, B. Emergence of Nanoplastic in the Environment and Possible Impact on Human Health. Environ. Sci. Technol. 2019 , 53 (4), 1748–1765. https://doi.org/10.1021/acs.est.8b05512. Kumar, R.; Manna, C.; Padha, S.; Verma, A.; Sharma, P.; Dhar, A.; Ghosh, A.; Bhattacharya, P. Micro(Nano)Plastics Pollution and Human Health: How Plastics Can Induce Carcinogenesis to Humans? Chemosphere 2022 , 298 , 134267. https://doi.org/10.1016/j.chemosphere.2022.134267. Zhang, H.; Zhang, S.; Duan, Z.; Wang, L. Pulmonary Toxicology Assessment of Polyethylene Terephthalate Nanoplastic Particles in Vitro. Environment International 2022 , 162 , 107177. https://doi.org/10.1016/j.envint.2022.107177. Materić, D.; Kasper-Giebl, A.; Kau, D.; Anten, M.; Greilinger, M.; Ludewig, E.; van Sebille, E.; Röckmann, T.; Holzinger, R. Micro- and Nanoplastics in Alpine Snow: A New Method for Chemical Identification and (Semi)Quantification in the Nanogram Range. Environ. Sci. Technol. 2020 , 54 (4), 2353–2359. https://doi.org/10.1021/acs.est.9b07540. Fraissinet, S.; De Benedetto, G. E.; Malitesta, C.; Holzinger, R.; Materić, D. Microplastics and Nanoplastics Size Distribution in Farmed Mussel Tissues. Commun Earth Environ 2024 , 5 (1), 1–8. https://doi.org/10.1038/s43247-024-01300-2. Cappellin, L.; Karl, T.; Probst, M.; Ismailova, O.; Winkler, P. M.; Soukoulis, C.; Aprea, E.; Märk, T. D.; Gasperi, F.; Biasioli, F. On Quantitative Determination of Volatile Organic Compound Concentrations Using Proton Transfer Reaction Time-of-Flight Mass Spectrometry. Environ. Sci. Technol. 2012 , 46 (4), 2283–2290. https://doi.org/10.1021/es203985t. Holzinger, R.; Acton, W. J. F.; Bloss, W. J.; Breitenlechner, M.; Crilley, L. R.; Dusanter, S.; Gonin, M.; Gros, V.; Keutsch, F. N.; Kiendler-Scharr, A.; Kramer, L. J.; Krechmer, J. E.; Languille, B.; Locoge, N.; Lopez-Hilfiker, F.; Materić, D.; Moreno, S.; Nemitz, E.; Quéléver, L. L. J.; Sarda Esteve, R.; Sauvage, S.; Schallhart, S.; Sommariva, R.; Tillmann, R.; Wedel, S.; Worton, D. R.; Xu, K.; Zaytsev, A. Validity and Limitations of Simple Reaction Kinetics to Calculate Concentrations of Organic Compounds from Ion Counts in PTR-MS. Atmospheric Measurement Techniques 2019 , 12 (11), 6193–6208. https://doi.org/10.5194/amt-12-6193-2019. Hansel, A.; Jordan, A.; Holzinger, R.; Prazeller, P.; Vogel, W.; Lindinger, W. Proton Transfer Reaction Mass Spectrometry: On-Line Trace Gas Analysis at the Ppb Level. International Journal of Mass Spectrometry and Ion Processes 1995 , 149–150 , 609–619. https://doi.org/10.1016/0168-1176(95)04294-U. Garcia, M. A.; Liu, R.; Nihart, A.; El Hayek, E.; Castillo, E.; Barrozo, E. R.; Suter, M. A.; Bleske, B.; Scott, J.; Forsythe, K.; Gonzalez-Estrella, J.; Aagaard, K. M.; Campen, M. J. Quantitation and Identification of Microplastics Accumulation in Human Placental Specimens Using Pyrolysis Gas Chromatography Mass Spectrometry. Toxicological Sciences 2024 , 199 (1), 81–88. https://doi.org/10.1093/toxsci/kfae021. Marfella Raffaele; Prattichizzo Francesco; Sardu Celestino; Fulgenzi Gianluca; Graciotti Laura; Spadoni Tatiana; D’Onofrio Nunzia; Scisciola Lucia; La Grotta Rosalba; Frigé Chiara; Pellegrini Valeria; Municinò Maurizio; Siniscalchi Mario; Spinetti Fabio; Vigliotti Gennaro; Vecchione Carmine; Carrizzo Albino; Accarino Giulio; Squillante Antonio; Spaziano Giuseppe; Mirra Davida; Esposito Renata; Altieri Simona; Falco Giovanni; Fenti Angelo; Galoppo Simona; Canzano Silvana; Sasso Ferdinando C.; Matacchione Giulia; Olivieri Fabiola; Ferraraccio Franca; Panarese Iacopo; Paolisso Pasquale; Barbato Emanuele; Lubritto Carmine; Balestrieri Maria L.; Mauro Ciro; Caballero Augusto E.; Rajagopalan Sanjay; Ceriello Antonio; D’Agostino Bruno; Iovino Pasquale; Paolisso Giuseppe. Microplastics and Nanoplastics in Atheromas and Cardiovascular Events. New England Journal of Medicine 2024 , 390 (10), 900–910. https://doi.org/10.1056/NEJMoa2309822. Oslakovic, C.; Cedervall, T.; Linse, S.; Dahlbäck, B. Polystyrene Nanoparticles Affecting Blood Coagulation. Nanomedicine: Nanotechnology, Biology and Medicine 2012 , 8 (6), 981–986. https://doi.org/10.1016/j.nano.2011.12.001. Sanfins, E.; Augustsson, C.; Dahlbäck, B.; Linse, S.; Cedervall, T. Size-Dependent Effects of Nanoparticles on Enzymes in the Blood Coagulation Cascade. Nano Lett. 2014 , 14 (8), 4736–4744. https://doi.org/10.1021/nl501863u. Nikpay, M. Polystyrene and Polymethylmethacrylate Microplastics Embedded in Fat, Oil, and Grease (FOG) Deposits of Sewers. Pollution 2022 , 8 (4), 1338–1347. https://doi.org/10.22059/poll.2022.342517.1464. Fraissinet, S.; Pennetta, A.; Rossi, S.; De Benedetto, G. E.; Malitesta, C. Optimization of a New Multi-Reagent Procedure for Quantitative Mussel Digestion in Microplastic Analysis. Marine Pollution Bulletin 2021 , 173 , 112931. https://doi.org/10.1016/j.marpolbul.2021.112931. Schneider, C. A.; Rasband, W. S.; Eliceiri, K. W. NIH Image to ImageJ: 25 Years of Image Analysis. Nat Methods 2012 , 9 (7), 671–675. https://doi.org/10.1038/nmeth.2089. Materić, D.; Peacock, M.; Kent, M.; Cook, S.; Gauci, V.; Röckmann, T.; Holzinger, R. Characterisation of the Semi-Volatile Component of Dissolved Organic Matter by Thermal Desorption – Proton Transfer Reaction – Mass Spectrometry. Scientific Reports 2017 , 7 (1), 15936. https://doi.org/10.1038/s41598-017-16256-x. Holzinger, R. PTRwid: A New Widget Tool for Processing PTR-TOF-MS Data. Atmos. Meas. Tech. 2015 , 8 (9), 3903–3922. https://doi.org/10.5194/amt-8-3903-2015. Materić, D.; Ludewig, E.; Xu, K.; Röckmann, T.; Holzinger, R. Brief Communication: Analysis of Organic Matter in Surface Snow by PTR-MS – Implications for Dry Deposition Dynamics in the Alps. The Cryosphere 2019 , 13 (1), 297–307. https://doi.org/10.5194/tc-13-297-2019. Peacock, M.; Materić, D.; Kothawala, D. N.; Holzinger, R.; Futter, M. N. Understanding Dissolved Organic Matter Reactivity and Composition in Lakes and Streams Using Proton-Transfer-Reaction Mass Spectrometry (PTR-MS). Environ. Sci. Technol. Lett. 2018 , 5 (12), 739–744. https://doi.org/10.1021/acs.estlett.8b00529. Yao, P.; Holzinger, R.; Materić, D.; Oyama, B. S.; de Fátima Andrade, M.; Paul, D.; Ni, H.; Noto, H.; Huang, R.-J.; Dusek, U. Methylsiloxanes from Vehicle Emissions Detected in Aerosol Particles. Environ. Sci. Technol. 2023 , 57 (38), 14269–14279. https://doi.org/10.1021/acs.est.3c03797. Yao, P.; Chianese, E.; Kairys, N.; Holzinger, R.; Materić, D.; Sirignano, C.; Riccio, A.; Ni, H.; Huang, R.-J.; Dusek, U. A Large Contribution of Methylsiloxanes to Particulate Matter from Ship Emissions. Environment International 2022 , 165 , 107324. https://doi.org/10.1016/j.envint.2022.107324. Materić, D.; Peacock, M.; Dean, J.; Futter, M.; Maximov, T.; Moldan, F.; Röckmann, T.; Holzinger, R. Presence of Nanoplastics in Rural and Remote Surface Waters. Environ. Res. Lett. 2022 , 17 (5), 054036. https://doi.org/10.1088/1748-9326/ac68f7. Ellis, A. M.; Mayhew, C. A. Proton Transfer Reaction Mass Spectrometry: Principles and Applications , 1 edition.; Wiley-Blackwell: Chichester, West Sussex, 2014. Thompson, K. M.; Burmaster, D. E.; Crouch3, E. A. C. Monte Carlo Techniques for Quantitative Uncertainty Analysis in Public Health Risk Assessments. Risk Analysis 1992 , 12 (1), 53–63. https://doi.org/10.1111/j.1539-6924.1992.tb01307.x. Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryInformationTablesandFiguresI.docx Additional Tables and Figures SuplementaryInformationIIQCandstatisticsanonimus.xlsx QC/QA and statistics SupplementaryInformationIIITDPTRMSDataanalysisandprocessinganonimus.xlsx TD-PTR-MS Data analysis suplement Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6154338","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":427866893,"identity":"fa700bc4-55fe-4b41-8c27-d6db407b4ef7","order_by":0,"name":"Dušan Materić","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5UlEQVRIiWNgGAWjYDCCAyjsCgZ+IG1AnBYeMPsMg2QDSVoYGNuI0MJ3I/3h5wKGbfL27L0PD/ycd1hCvoF54wN8WiRv5BhLz2C4bdjDc9zgYO+2wxKMDWzFeK0xuJHDIM3DcJuxRyKN4TDjtsN1zAw8ZhL4taQ//g3UYt8j/wyoZc5hCTYGHvMf+LUkmIFsSewBqj3M2HBYggdoCz4dDJJn3phZ8xjcTu45k8ZwsOdYuoQEM1sxXofxHU9/fJun4rZte/sx5g8/aqwl5NubN37Aaw3EecgcZsLqR8EoGAWjYBQQAAC0wUbl07HmAAAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-6454-3456","institution":"Helmholtz Centre for Environmental Research - UFZ; Department of Environmental Analytical Chemistry","correspondingAuthor":true,"prefix":"","firstName":"Dušan","middleName":"","lastName":"Materić","suffix":""},{"id":427866894,"identity":"d391b9c6-7150-4e4f-bd70-8abf19c084ab","order_by":1,"name":"Vaishnavi Tokla","email":"","orcid":"","institution":"Helmholtz Centre for Environmental Research - UFZ; Department of Environmental Analytical Chemistry","correspondingAuthor":false,"prefix":"","firstName":"Vaishnavi","middleName":"","lastName":"Tokla","suffix":""},{"id":427866895,"identity":"dea5b8e1-158c-4e35-9710-70ae591b9b4e","order_by":2,"name":"Tommy Cedervall","email":"","orcid":"https://orcid.org/0000-0003-2255-8446","institution":"Lunds universitet","correspondingAuthor":false,"prefix":"","firstName":"Tommy","middleName":"","lastName":"Cedervall","suffix":""},{"id":427866896,"identity":"bb05f020-86d2-4b56-a5bc-3dac53887b6d","order_by":3,"name":"Rupert Holzinger","email":"","orcid":"https://orcid.org/0000-0003-1902-1824","institution":"Universiteit Utrecht","correspondingAuthor":false,"prefix":"","firstName":"Rupert","middleName":"","lastName":"Holzinger","suffix":""},{"id":427866897,"identity":"52b3f87c-f788-433d-b783-72ac3cbcaccb","order_by":4,"name":"Mikael Ekvall","email":"","orcid":"","institution":"Biochemistry and Structural Biology, Lund University","correspondingAuthor":false,"prefix":"","firstName":"Mikael","middleName":"","lastName":"Ekvall","suffix":""},{"id":427866898,"identity":"1b003337-ac2f-4de1-9c71-9000387e5c11","order_by":5,"name":"Maria Värendh","email":"","orcid":"","institution":"Lund University","correspondingAuthor":false,"prefix":"","firstName":"Maria","middleName":"","lastName":"Värendh","suffix":""}],"badges":[],"createdAt":"2025-03-04 12:10:27","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6154338/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6154338/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":79569471,"identity":"6a60e429-7475-49da-9e7c-b276da54ef52","added_by":"auto","created_at":"2025-03-31 10:12:10","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":104904,"visible":true,"origin":"","legend":"\u003cp\u003eSize distribution of micro and nanoplastics in tonsil samples with each polymer detected. The mean mass of micro-and nanoplastics from all tonsil samples according to sizes classes (a). The mass of the plastic particles is defined in nanogram/milligram on the y-axis. The percentage of different types of plastics in the 20-200 nm (nanoplastics, NPs) (b), and \u0026gt;2700 nm (microplastics, MPs) (c). All values are corrected for measured blank contamination. The error bars represent the standard deviation of triplicates. The plastic types are Tyre, PET- Polyethylene terephthalate, PS - Polystyrene, PVC - Polyvinyl Chloride, PP – Polypropylene, PPC- Polypropylene Carbonate, PE – Polyethylene.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/ebf7d600576b68f0d0dcab7f.png"},{"id":79569476,"identity":"5a9dfd0d-92c8-4048-b3a8-091d7cee7778","added_by":"auto","created_at":"2025-03-31 10:12:11","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":66574,"visible":true,"origin":"","legend":"\u003cp\u003eNanoplastic and microplastic concentrations for each patient and plastic-type (size groups for nanoplastics 20-200 nm and microplastic \u0026gt;2700nm, respectively). Full details on size fractions charts can be found in supplement (Figure S1). All values are corrected for measured blank contamination. The error bars represent the standard deviation of triplicates. The plastic types are Tyre, PET- Polyethylene terephthalate, PS - Polystyrene, PVC - Polyvinyl Chloride, PP – Polypropylene, PPC- Polypropylene Carbonate, PE – Polyethylene.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/b4bfe0e6fcb7f00e492e0999.png"},{"id":79569813,"identity":"2067025e-6c22-4f38-b8d4-7113b28aa4dd","added_by":"auto","created_at":"2025-03-31 10:20:11","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":156285,"visible":true,"origin":"","legend":"\u003cp\u003eThe mass and size distribution from tonsils coming from different donors. Patient 5 (a), patient 6 (b), Patient 15 (c), Patient 3 (d) (For all the patients, see Figure S2). The plastic types are Tyre, PET- Polyethylene terephthalate, PS - Polystyrene, PVC - Polyvinyl Chloride, PP – Polypropylene, PPC- Polypropylene Carbonate, PE – Polyethylene. Error bars represent the standard deviation of triplicates.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/91705678aacdf14bc2e65459.png"},{"id":79569478,"identity":"85ed0cf0-2142-4ea5-a01a-4abc280ecc75","added_by":"auto","created_at":"2025-03-31 10:12:11","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":36423,"visible":true,"origin":"","legend":"\u003cp\u003eIdentification of tyre particle in human tonsil in TD-PTR-MS. The top panel shows organic ions in the tyre wear library. The bottom panel shows tire-associated ions in an actual human tonsil sample after the successful digestion of the organic tissue (patient 8). Note the organic ions signal similarity (relative intensity and correlation) between the virgin polymer and the human tonsil sample. The match score had a p-value \u0026lt; 0.00032, indicating that the coincidence of random false identification is less than 0.032%. In 42 size groups out of 60 in total, we identified the tyre particles (Table S1).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/959cec7398c933cd06fe93b5.png"},{"id":79569480,"identity":"2ab466c2-e6a2-44c4-9e10-2c13e49aae15","added_by":"auto","created_at":"2025-03-31 10:12:11","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":57730,"visible":true,"origin":"","legend":"\u003cp\u003eMonte Carlo Assessment of Overestimation. Organic matter was in-silico increased step by step, randomly distributed across the 40 organic ions used for plastic identification (fingerprint), up to 350% (x-axis). Overestimation (y-axis) did not exceed 31% (indicated by the dashed horizontal line). Notably, overall quantification dropped after 100% (for PVC and TWP) and 200% (for PET) organic matter increase, demonstrating that the 40-ion fingerprinting approach effectively prevents severe overestimation. Abbreviations: OM – organic matter, PET – Polyethylene terephthalate, PVC – Polyvinyl chloride, TWP – Tyre wear particles.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/9471063870167c73b529445c.png"},{"id":79571809,"identity":"a7c3b7f6-fb7a-42ce-ba5d-4aeed26d99b7","added_by":"auto","created_at":"2025-03-31 10:44:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1077338,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/953472ad-9290-4d36-8b63-793197ad07af.pdf"},{"id":79569472,"identity":"70fb6b7f-2fff-4584-b05b-ae21df00b669","added_by":"auto","created_at":"2025-03-31 10:12:11","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":261737,"visible":true,"origin":"","legend":"Additional Tables and Figures","description":"","filename":"SupplementaryInformationTablesandFiguresI.docx","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/33dbf16672a02b4ab6900b7d.docx"},{"id":79570508,"identity":"941c8f78-6821-4be4-ae67-977d64ce50a3","added_by":"auto","created_at":"2025-03-31 10:28:11","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":47234,"visible":true,"origin":"","legend":"QC/QA and statistics","description":"","filename":"SuplementaryInformationIIQCandstatisticsanonimus.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/96df6a4f9c38224c464f83f7.xlsx"},{"id":79569817,"identity":"7c573756-8e7a-4a46-bc92-5d36e82e4bce","added_by":"auto","created_at":"2025-03-31 10:20:11","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":343915,"visible":true,"origin":"","legend":"TD-PTR-MS Data analysis suplement","description":"","filename":"SupplementaryInformationIIITDPTRMSDataanalysisandprocessinganonimus.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-6154338/v1/90ec78d3101021e473286271.xlsx"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Size distribution of nanoplastics and tyre wear particles in human tonsils","fulltext":[{"header":"Introduction","content":"\u003cp\u003ePlastic is considered the most widely used material in the world, as only in Europe the total plastics production reached 54 million tons in 2023\u003csup\u003e1,2\u003c/sup\u003e. The worldwide production of plastic has grown exponentially in just a few decades – from 2 Mt in 1950 to 413.8 Mt in 2023 – and with it, the amount of plastic waste\u003csup\u003e2\u003c/sup\u003e. Historically, plastics have attracted considerable attention due to their low production costand favourable characteristics, including light weight, durability, flexibility, and water resistance. However, the same attributes that make plastics desirable—especially durability and resistance to degradation—also naturally contribute to their long-term persistence in the environment. Thus, plastics are not easily biodegradable and can survive for centuries\u003csup\u003e3\u003c/sup\u003e, and their pollution is recognised to be one of the greatest environmental challenges of the 21st century: they might cause wide-ranging damage to ecosystems\u003csup\u003e4\u003c/sup\u003e and human health\u003csup\u003e5–7\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePlastics are made from synthetic or semisynthetic polymers, which consist of long, repeating chains. Various additives, such as plasticisers, flame retardants, and colourants, are often incorporated to improve their properties and appearance\u003csup\u003e8–10\u003c/sup\u003e. In Europe, the most widely used plastics are polypropylene (PP) and polyethylene (PE), including its low-density form (LDPE), followed by polyvinyl chloride (PVC), polyethylene terephthalate (PET), and polystyrene (PS)\u0026nbsp;\u003csup\u003e2\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eThe growing environmental consequences of plastic pollution have been topics of considerable interest in recent years, especially small plastic particles known as microplastics and nanoplastics. Microplastics are generally defined as plastic particles that are less than 5 mm in size, whereas nanoplastics are even smaller, these are often defined as plastic particles that are less than 1 µm in size\u0026nbsp;\u003csup\u003e11,12\u003c/sup\u003e. These plastics are mostly formed from the breakdown of larger plastic items in the environment; however, many cosmetics and skincare products contain intentionally produced and added microbeads, which, to a lesser extent, contribute to overall environmental microplastic pollution\u003csup\u003e13\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eMicro- and nanoplastics are ubiquitous. Studies have found microplastic particles in various environmental compartments. For instance, microplastics have been found in river sediments\u0026nbsp;\u003csup\u003e14,15\u003c/sup\u003e, agricultural soil\u003csup\u003e16\u003c/sup\u003e, oceans\u0026nbsp;\u003csup\u003e17–20\u003c/sup\u003e and even in remote regions such as the deep sea\u003csup\u003e21,22\u003c/sup\u003e or the Antarctic ice\u003csup\u003e23–25\u003c/sup\u003e. The detection and characterisation of nanoplastics presents technical challenges\u003csup\u003e26,27\u003c/sup\u003e. However, there is a limited number of studies indicating that nanoplastics can be found wherever microplastics are detected\u003csup\u003e28\u003c/sup\u003e. Correspondingly, nanoplastics have been found, for example, in the Dutch Wadden sea\u003csup\u003e20\u003c/sup\u003e, southern and northern polar ice\u003csup\u003e29\u003c/sup\u003e, and in alpine snow and glaciers\u003csup\u003e30,31\u003c/sup\u003e, atmospheric fallouts\u003csup\u003e32\u003c/sup\u003e and urban\u003csup\u003e33\u003c/sup\u003e, rural and remote air\u003csup\u003e34\u003c/sup\u003e. The ubiquitous occurrence of micro- and nanoplastics, even in remote regions such as the deep sea or Antarctica, shows that these particles are transported globally.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn addition to the inanimate environmental samples, microplastic particles have also been found in a variety of organisms and human tissues. In humans, microplastics have been detected in blood,\u003csup\u003e35\u003c/sup\u003e liver,\u003csup\u003e36\u003c/sup\u003e lung,\u003csup\u003e37\u003c/sup\u003e kidney and brain tissue\u0026nbsp;\u003csup\u003e38\u003c/sup\u003e as well as throughout the reproductive system, including semen,\u003csup\u003e39\u003c/sup\u003e testes,\u003csup\u003e40,41\u003c/sup\u003e uterus,\u003csup\u003e42\u003c/sup\u003e and placenta.\u003csup\u003e43\u003c/sup\u003e While their effects have not yet been fully characterised, exposure to plastics has been linked to cell apoptosis and both genotoxicity and cytotoxicity.\u003csup\u003e44–46\u003c/sup\u003e Additionally, plastic particles may release plasticisers/additives and adsorbed pollutants, potentially enhancing their effects\u003csup\u003e9,10\u003c/sup\u003e. The\u0026nbsp;prevalence of small plastic particles in human beings is somewhat known, but no study has shown nanosized particles originating from tyres in human organs. Nanoplastics and tyre wear particles have the potential to cause significant impact due to their small size and large surface area. However, the effects of these widely dispersed nanoparticles on human health have yet to be thoroughly investigated.\u003c/p\u003e\n\u003cp\u003eIn humans, the tonsils are the first immunologically active tissue that encounters particles upon oral and nasal exposure. Tonsils are located in the oral cavity, and air, food and liquid pass the tonsils on route to the respiratory and digestive tracts. Their position optimises the uptake of foreign materials like antigens and possibly micro- and nanoplastics. In this work, we aimed to quantify presence and size distribution and types of micro- and nanoplastics in tonsil tissues and hypothesised that tonsils, besides skin, digestive and respiratory systems, could play an active and significant role in human micro- and nanoplastics exposure pathway.\u003c/p\u003e"},{"header":"Material \u0026 Methods","content":"\u003cp\u003eTonsil samples\u003c/p\u003e\n\u003cp\u003eThe tonsils were obtained from individuals who underwent conventional tonsillectomy at the Otorhinolaryngology department in Ystad, southern Sweden. The indications for tonsillectomy are sleep apnoea or chronic tonsilitis. The tonsils would, if not used in the study, have been destroyed. Prior to sampling, all patients signed informed consent. The consent form was signed by the legal guardians of underage patients (\u0026lt;18 years old). Following surgery, the tonsils were placed in 50 mL falcon tubes (Eppendorf, Sweden) and stored one night in a freezer and then delivered to the Department of Otorhinolaryngology in Lund, southern Sweden, and then further on to the Department of Biochemistry and Structural Biology at Lund University, where they were stored at -20 \u0026deg;C until further analysis began. The study was approved by The Regional Ethical Committee (Dnr 2018/611) and the Regional Bioethical Committee (136/BD16).\u003c/p\u003e\n\u003cp\u003eThere were 15 tonsils included in the study from 15 different patients with a mean age of 18 years (min-max 3-44 years), of which 9 were female and 8 were children (\u0026lt;18 years). The cohort was mainly healthy except for their problems with sleep apnoea and recurrent tonsillitis. Among the adults, two persons reported smoking, and they reported smoking\u0026gt;20 cigarettes per day. Ten of the patients were living in a city with more than 100000 inhabitants. It was common to drink and eat from plastic containers; 10 persons were drinking from a plastic bottle more than twice a week, and six had a daily drink from a bottle made of plastic. Eating from plastic containers daily was reported by 6, and at least twice a week by 12 persons. This data is summarised in Table S2.\u003c/p\u003e\n\u003cp\u003eChemicals and materials\u003c/p\u003e\n\u003cp\u003ePotassium hydroxide (KOH) pellets were obtained from Sigma-Aldrich, and a hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, 3%) solution was supplied by Frost Pharma. All the solutions were prepared using HPLC water (VWR, Germany), and the digestion solution was freshly prepared daily. Glass fibre filters were purchased from Whatman\u003csup\u003eTM\u003c/sup\u003e, while quartz fibre filters, anodisc filters, and the glass filtration setup were supplied by Sterlitech. The isolation of plastic particles from the tonsil tissue was evaluated using polystyrene particles of 0.5 \u0026mu;m size, sourced from Sigma-Aldrich. These particles were used to spike samples for recovery experiments. The same chemicals and materials are used for the samples and complete procedural blanks.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSample preparation\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe tonsil tissues were removed from the freezer and rinsed four times with Milli-Q water to remove any residual blood from the surgery. The tissues were then freeze-dried for 24 hours at \u0026minus;55\u0026deg;C, grounded using a ceramic mortar, and weighed. To prevent contamination and rehydration, the tonsil tissues were stored in a desiccator and wrapped in aluminium foil.\u003c/p\u003e\n\u003cp\u003eThe digestion and extraction of nanoplastics were performed with slight modifications to the previously published protocol\u003csup\u003e48,57\u003c/sup\u003e. Tonsil tissue samples of a known quantity of 10 mg were transferred into glass bottles. To each bottle, 8 mL of digestion solution consisting of 2.5% KOH and 3% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in a 1:1 ratio was added. The bottles were then incubated in a water bath at 60\u0026deg;C for 8 hours to facilitate the breakdown of organic matter. After incubation, the samples were allowed to cool to room temperature, and the solutions were neutralised by the dropwise addition of 10% citric acid until a neutral pH was achieved.\u003c/p\u003e\n\u003cp\u003eSolutions were filtered in cascade using different filter membranes and pore sizes including 2700 nm (Glass fibre filter), 1200 nm (Silver membrane filter), 200 nm (Anodisc filter), and 20 nm (Anodisc filter). Each bottle was rinsed twice with HPLC water. After each filtration step, the filters were washed with HPLC water to ensure no residual particles remained. Each filter was dried and stored in clean, sterile glass petri dishes for further analysis. Additionally, two procedural blanks were included to evaluate potential contamination and impurities arising from laboratory equipment and the procedural methodology.\u003c/p\u003e\n\u003cp\u003eFollowing the filtration, the filters were dried and cut in a laminar flow workstation. For the glass fibre and silver membrane filters, cutting was performed using a metal punch tool with a 3 mm diameter. The anodisc filters (200 nm and 20 nm pore size) were cracked using a spatula and tweezers. The resulting filter pieces were photographed, and their surface areas were estimated using ImageJ software\u003csup\u003e58\u003c/sup\u003e. Tools and surfaces were cleaned with ethanol and cellulose tissue between uses. Three aliquots from each filter stage of each sample and blank were then transferred into pre-baked (250\u0026deg;C, overnight) 10 mL glass vials.\u003c/p\u003e\n\u003cp\u003eTD-PTR-MS Analysis\u003c/p\u003e\n\u003cp\u003eThe samples were analysed using thermal desorption proton transfer mass spectrometry (TD-PTR-MS), as previously described in detail\u003csup\u003e47,59\u003c/sup\u003e. Briefly, the TD-PTR-MS protocol was carried with the following parameters: a temperature ramp from 35\u0026deg;C to 360\u0026deg;C, an E/N ratio of 120 Td, and a p-drift pressure of 2.9 mbar to minimise clustering and fragmentation. TD-PTR-MS analysis was performed using a PTR8000 instrument (Ionicon Analitik, AU). Both samples and blanks were analysed in triplicates to ensure data reliability and reproducibility.\u003c/p\u003e\n\u003cp\u003eThe first level of data processing (peak identification, integration, TOF calibration, ion quantification) was conducted using the PTRwid software\u003csup\u003e60\u003c/sup\u003e. The TD-PTR-MS signal was integrated over a 7-minute period, starting when the thermal desorption temperature reached 200\u0026deg;C, to capture the thermal degradation products of each polymer. The mass spectra were corrected by subtracting the blank signal, and organic ions below the 3\u0026sigma; detection limit were excluded from further analysis.\u003c/p\u003e\n\u003cp\u003eTD-PTR-MS has been applied for the analysis of organic aerosols\u003csup\u003e61\u003c/sup\u003e, dissolved organic matter (DOM)\u003csup\u003e62\u003c/sup\u003e, Methylsiloxanes\u003csup\u003e63,64\u003c/sup\u003e, and nanoplastics in different environments, including seawater\u003csup\u003e20\u003c/sup\u003e, surface water\u003csup\u003e65\u003c/sup\u003e, snow\u003csup\u003e47\u003c/sup\u003e, ice\u003csup\u003e29\u003c/sup\u003e, glaciers\u003csup\u003e31\u003c/sup\u003e, atmospheric samples\u003csup\u003e33,34\u003c/sup\u003e, and animal tissue\u003csup\u003e48\u003c/sup\u003e. Similarly, here, we obtained plastic identification via the fingerprinting approach, which utilises the 40 most abundant library ions with m/z \u0026gt;100 for the identification and quantification of microplastics and nanoplastics in the samples. Fingerprinting was performed on all samples and blanks (see Data Availability section for all the steps of the analysis). In short, the ion signals of each sample were compared to those in the reference library to identify matching patterns (Figure 4). The similarity was assessed based on the ratio of ion signals from the library using four different matching algorithms. If a match was obtained (z-score \u0026gt; 2), each identified ion present in the plastics\u0026mdash;after contamination correction\u0026mdash;was quantified following the standard PTR-MS protocol\u003csup\u003e49,50\u003c/sup\u003e. Details on data processing and matching algorithms can be found in previous studies, e.g.\u003csup\u003e20,47,48\u003c/sup\u003e, and the standard PTR-MS quantification method is explained elsewhere\u003csup\u003e49\u0026ndash;51,66\u003c/sup\u003e. A step-by-step outline of our data analysis and processing, from raw files to final results, is provided in the supplementary information and Data Availability section.\u003c/p\u003e\n\u003ch2\u003eQuality Control and Assurance (QC/QA)\u003c/h2\u003e\n\u003cp\u003eGiven that the presence of plastics is unavoidable, extra care was taken to prevent contamination of the samples. A laminar flow was used to avoid contamination from the surroundings. Laboratory staff wore cotton lab coats and nitrile gloves. Additional precautions were taken to minimise contamination, such as using silicone and Teflon materials, with silicone vacuum tubing and Teflon discs replaced on the glass vial seals. All glassware was heat-treated at 250\u0026deg;C overnight, and all containers were covered with aluminium foil to prevent airborne plastic contamination. Procedural blanks were included to assess potential contamination from the laboratory environment, reagents, or equipment.\u003c/p\u003e\n\u003cp\u003eOur experiments included three different blanks:\u003c/p\u003e\n\u003cp\u003ea) Procedural blanks to evaluate contamination during laboratory operations. These represent control samples treated the same way as the actual tonsil samples but without tonsil tissue, allowing for the identification of any plastics introduced by the laboratory environment or procedures.\u003c/p\u003e\n\u003cp\u003eb) Filter blanks to measure possible contamination from the filter material.\u003c/p\u003e\n\u003cp\u003ec) System blanks to assess contamination from the instrument.\u003c/p\u003e\n\u003cp\u003eIn the procedural blanks, some traces of plastic were detected. For example, extraction batch that included patient 7 and associated procedural blanks, we detected an average of 3.7 ng absolute of tyre particles in the procedural blanks, while the sample itself contained 37\u0026ndash;707 ng absolute (see Supplementary Information and Data Availability for details). Furthermore, the results showed that the plastic content in the samples was significantly higher than in the procedural blanks for all plastic types, indicating that most of the plastics detected\u0026mdash;such as tyre particles, PET, PS, PVC, PP/PPC, and PE\u0026mdash;originated from the samples rather than from contamination during the experimental process. This confirms the presence of various plastics in the actual samples, particularly PVC and PP/PPC, which were absent in the blanks for nanoplastics. All reported values were corrected using the mean blank value for each extraction batch.\u003c/p\u003e\n\u003cp\u003eThe tonsils samples were collected specifically for the project and transport from surgery theatre has been planned to reduce contamination with short way from patient into sample storing container. Potential contamination is still possible and it was experimentally reduced by washing the organs and avoiding sampling from the tissue surface, which could have been exposed to external contaminants. However, we acknowledge that the best blank control would have been archived tonsil samples from the pre-plastic era, but we did not have access to such specimens.\u003c/p\u003e\n\u003cp\u003eAdditionally, to evaluate the ionisation efficiency and recovery rate of our analytical method, we performed spiking assessments on both procedural blanks and sample filters by adding a known quantity of PS particles to random runs. The ionisation efficiency and recovery rate averaged 30%, which is similar to previously obtained values\u003csup\u003e31,48\u003c/sup\u003e. Furthermore, spiking the samples with PS showed good match scores (z-score \u0026gt; 4 for at least one algorithm), suggesting that no matrix interference (e.g., from insufficient tissue digestion) compromised the plastic fingerprinting process.\u003c/p\u003e\n\u003cp\u003eIdeally, further analysis of such samples, besides TD-PTR-MS, could be coupled with optical methods that facilitate the chemical identification of polymers and have the same or a similar analytical window, such as Raman spectroscopy. TD-PTR-MS and Raman have previously been used in tandem, yielding overall similar results\u003csup\u003e32\u003c/sup\u003e; however, this was achieved for relatively clear matrices, such as atmospheric deposition. In this project, which involved human samples, such cross-comparison of different methods was not technically possible. Developing a digestion protocol and a pre-concentration method compatible with both Raman spectroscopy and TD-PTR-MS should be a future focus of analytical development to improve QC/QA.\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003eAssessing the uncertainties\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eIn this study, we performed replicated measurements to assess the experimental uncertainty in our analysis of micro- and nanoplastics. We also included procedural blanks to apply suitable corrections for any plastics that might have been introduced during sample handling. However, additional factors could contribute to both underestimation and overestimation. Here, we outline the potential causes and assess the associated uncertainties.\u003c/p\u003e\n\u003ch4\u003e\u003cstrong\u003eUnderestimation\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eThe most prominent factor leading to underestimation is related to ionisation efficiency and recovery in proton transfer reaction mass spectrometry. We tested ionisation efficiency and recovery by spiking blanks with a known amount of an analytical-grade PS standard, obtaining an average recovery of 29.7% (see SI). However, quantitative analytical standards for other nanoplastics do not exist, and we cautiously assume that this value holds, on average, for other plastic types.\u003c/p\u003e\n\u003cp\u003eAdditionally, we account for the PTR-MS uncertainty of \u0026plusmn;30%\u003csup\u003e50\u003c/sup\u003e. Using these two parameters, we calculate the underestimation factor as:\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAOAAAAA6CAYAAACpiFWoAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAAZfSURBVHhe7dzPaxPNAwbwZ9+7Nok5Fekh21uFgoSeeunBbNCjSmIUTyJsboKpEPUkHjYqgvSQgIInG8Eet7QVetnQg1VIoOLF+BfsotU/YN7DtzNkJ43t127etdvnA8FkJpuOkCc7v3YNIYQAEcXiH72AiP47DCBRjBhAohgxgEQxYgCJYsQAEsWIASSKEQNIFCMGkChGDCBRjBhAohgxgEQxYgCJYsQAEsWIASSKUWIDuLq6inK5rBcjCAIUi0UYhoHp6Wmsrq6G6nu9Hqanp2EYBorFInq9XqieKEqJDOD9+/dx6dIl/PjxQ6/Cw4cPkUqlIIRArVbDjRs38O3bN1V/+fJl1Go1CCGQSqVw69at0PFEkRIJ4ziO8DxPOI4jLMsK1fm+LwCIbrerytLptGg2m0IIIVzXFel0WtV1u92h9xNFKXFnwMXFRczPz+vFAIAvX74AAGZnZ1XZ3Nwcdnd3AQA7OzuYm5tTdfJ9v379UmVEUUpcAP/E5uamXhSytbWlFxFFggEEkEql9KKQiYkJvYgoEgwggPPnz+tFITMzM3oRUSROVAAnJycBAJ1OR5V9+PABU1NTAICpqSmsr6+rOrkEIY8jilpiAygnVgblcjlYloWlpSUAQLvdBgBcuHBB/ZtOp9FqtQAArVYLlmUhl8sNfApRhPRp0eNueXlZ5PN5AUAAEPl8XiwvL6v6brcrLMsSAIRpmkNLDK7rCtM0BQBhWZbwfT9UTxQlQ/DO2ESxSWwX9G/XaDTUlrf9tsS1221kMhkYhoFqtYogCEJ18jh9q1yj0UCj0QiVRemgdktyu5/+GNx1RAnsgsbJcRzhOI5ePETu0vF9X/i+L0qlUmgHTr/fVztwfN8XpmmKer2u6kzTFL7vi2azKUqlUui4fD4/tm7zQe0epHff6/V6qK30PwxghA4bQJ3jOMI0TfVa/7I2m031Rfc8T9i2LcTe1rrB7Xa2bYfGu+Omt/t30um0cF1XLz7x2AWNURAEaLfb2NzcxMrKiir/9OlTaG1yZmYG379/V6+lM2fOqOe9Xg/b29v7XgEStVHtHkV2py9evKhXnXgMYEw6nQ6y2SyuXbuGXC6Hs2fP6m8Z0ul0MDk5iY2NDQRBgFarpZZI7t27h+fPn+uH7GvU+Ew+isWifojyJ+1+/fo1bt++rRcTOAY8Es/z1HLHqIfnefphIXLcNtjltCwr1JWVf0cumdTrdbWM0u/3hed5qisq68Y5FhQj2r0feUVJv9/XqyhJY0D9iz+ux+/86RhQBkwaFcBRZBBd11WBsG37j9ry/zioXWKvHQeF9CRLTBd078dk7I9xSafT6nkulwtdofH582eYpqleD2q32ygUCsjlctjZ2VFjx0qlMvIqj6N0QXWD7dbJseLNmzf1KtqTmAAeJ41GA+VyGUEQIAgCLC0thSZPKpUK1tfX0ev1EAQBXr58ue8YKggCPHv2DHfv3tWrgN9c5bG2tjb0wzL4WFtb0w8BDtFuwzBCa5Dv37/n5MsBEhXAdrsdWiQeXKQOggDValX9yler1dCxg/Qzgvy8qFy5cgWZTAbZbBbZbBaZTAaPHj1S9fPz83AcBwsLC8hmsygUClhcXAx9BgC8evUKV69eVRMx586dU2e9ra0tLCwsaEcczUHt1nHy5RD0Pulx5TiOmniQi8SDa1S2bQvbtlV9Pp8fOUbK5/NDr+VtK/4WcoFen2iR+1z1hXD6OyUmgDrXdUMTBNDu7eK67lDQ9iNn8fhlpnFIVBd00M+fP4cmLk6dOqWenz59Gh8/fgzV76fVasG27dCiN1FUEhvABw8e4MWLF+q1aZp4+vQpsDcePMx9XuQsXqVS0auIIpHIAFarVZRKpdDs28rKCjY2NtQ0++7u7tAZUvfu3TtkMpmRd1kjOqrEBVBe5f748eNQ+ezsLL5+/QohBLa3tzExMYFCoRB6j+7Jkyeo1Wp6MVF09EHhceZ53qG2YMkZxN9tE5O7PA76LKKjSMwZMAgC3LlzB2/fvlUTJuVyeegC0F6vh+vXr6NQKKiuZafTgWEYoZs1vXnzhpMvNH56Io+rZrM5tG9T/+/Zti1M0xy6Zk6e7QbPiPpronHgPWGIYpSYLijRccQAEsWIASSKEQNIFCMGkChGDCBRjBhAohgxgEQxYgCJYsQAEsWIASSK0b/QcjFGnqcVsgAAAABJRU5ErkJggg==\"\u003e\u003c/p\u003e\n\u003cp\u003eThis suggests that our reported concentrations may be underestimated by a factor of 3.7.\u003c/p\u003e\n\u003ch4\u003e\u003cstrong\u003eOverestimation\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eOverestimation may occur if organic matter interferes with the ions used for identification and quantification. Such interference can arise, for example, from incomplete digestion of organic tissue. To estimate this effect, we performed a Monte Carlo simulation\u003csup\u003e67\u003c/sup\u003e to assess the impact of organic matter (OM) contamination on our 40 marker ions (see SI).\u003c/p\u003e\n\u003cp\u003eIn this simulation, we stepwise increased OM signal contributions by up to 350%, repeating each step 1000 times. The results, summarised in Figure 5, indicate that increased organic matter can lead to overestimation, but only up to 31%. Beyond this threshold, the 40-ion plastic fingerprinting approach prevents further overestimation.\u003c/p\u003e\n\u003cp\u003eFor example, when organic matter impurities increase by 200% over the 40 PET-associated ions, overestimation rises to 31%, but further increases in OM distort the mass spectra, preventing a positive PET match. This Monte Carlo analysis highlights the importance of a multi-ion approach, which is significantly more resistant to false positives than traditional methods relying on one or a few ions, such as py-GC-MS or TED-GC-MS.\u003c/p\u003e\n\u003cp\u003eUsing the Monte Carlo values and a PTR-MS uncertainty of 30%, we calculate total overestimation uncertainty as:\u003c/p\u003e\n\u003cp\u003e0.31 + (0.31\u0026times;30%) = 0.40\u003c/p\u003e\n\u003cp\u003eThis suggests that our overall overestimation could be up to 40%.\u003c/p\u003e"},{"header":"Results \u0026 Discussions","content":"\u003cp\u003eThis study shows the first data of micro- and nanoplastics as well as tyre size distribution in human tonsils. Our conservative approach to nanoplastic measurements relies on strict quality control of the analysis, including tested digestion protocol, multiple blanks (including complete procedural blanks), triplicate samples, spiking experiments with known amount of nanoplastics, conservative plastic detection using 40-ion fingerprinting and detection\u003csup\u003e47,48\u003c/sup\u003e for each plastic type and plastics quantification based on well-established PTR-TOF-MS quantification procedures\u003csup\u003e49\u0026ndash;51\u003c/sup\u003e widely applied in atmospheric, environmental, and food analysis. Details on QC/QA including detailed assessment of the uncertainties can be found in the Methods section.\u003c/p\u003e\n\u003cp\u003eAll 15 samples from different individuals showed the presence of at least one type of micro- and/or nanoplastics, spanning the mass concentration over four orders of magnitude (from 1 to 12000 ng/mg). The mean nanoplastics concentration for the 20-200 nm particle size was 350 ng/mg DW (median 126 ng/mg DW), reaching over 1200 ng/mg DW. Microplastic loads (size \u0026gt;2700 nm) were, on average, 2500 ng/mg (median 2083 ng/mg DW) and reached 9400 ng/g DW of tonsil tissue (Table S1).\u003c/p\u003e\n\u003cp\u003eThe average size distribution for all plastic types analysed in the samples is shown in Figure 1. For most of the samples (n=14), we measured the mass increase either on the 2700 nm filter or cut-off size fraction 20-200 nm or both. Comparatively, little plastic mass was observed in the size range of 200 \u0026ndash; 1200 nm, and 1200 -2700 nm. The primary polymer types for microplastics were tyre wear, PVC, and PS, while for nanoplastics, the major types were tyre wear, PE, and PVC (Figure 1b and c). To our knowledge, it is the first time tyre wear nanoparticles have been detected in human tissues. However, the presence of most polymer types was evident in all size ranges, but it was somewhat different for each sample (Figure 2).\u003c/p\u003e\n\u003cp\u003eWe did not observe the absolute dominance of the PE polymer as reported in previous studies for microplastics in human samples (including placenta, artery plaque, kidney, liver and brain)\u003csup\u003e38,52,53\u003c/sup\u003e. In fact, in 9 out of 15 samples (60%), we did not detect any PE nanoplastics, and in 10 out of 15 samples (different once than for PE), we did not detect PVC nanoplastics either. This discrepancy can be explained by methodological differences, as recent findings suggest that PE and PVC could be falsely assigned as present in py-GC-MS. In contrast, TD-PTR-MS, with its high resolution and 40-ion identification approach, provides much more conservative data for nanoplastics in complex matrices compared to py-GC-MS.\u003c/p\u003e\n\u003cp\u003eWe observe a considerable variation in plastic loads and types across different size fractions and patients (Figure 2). This significant variation in the load of specific plastic types and sizes is likely due to vast differences in individual exposure. Notably, some polymer types are absent in certain patients, while others appear only within specific size groups. This suggests that exposure to different plastic types may be influenced by particle size\u0026mdash;e.g., depending on the initial size distribution of plastics in air or food. These findings highlight the need for future systematic and comprehensive analysis of micro- and nanoplastic size distribution in indoor and occupational environments, as well as in food and beverages that humans are exposed to. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTo our knowledge, the quantity of nanoplastics and size distribution of the plastic for biological samples is only reported for seashells \u0026ndash; mussels\u003csup\u003e48\u003c/sup\u003e, and a similar analytical approach is used. The human tonsils contained 4 times more nanoplastics and 20 times more microplastics than the farmed mussels. Both these studies included a limited number of samples and showed a vast diversity of NPs concentrations. Sizes and types of micro- and nanoplastics vary from tonsil to tonsil in terms of concentration and plastic type, as exemplified in Figure 3. This additionally shows that human exposure might be highly diverse on a case-to-case basis. Using the same method, we previously measured tyre wear particles in farmed mussels, which contributed around 6% of total nanoplastics and only 1% of microplastics load, respectively\u003csup\u003e48\u003c/sup\u003e. However, for the human tonsil tissue, we observed a relative tyre wear contribution of 27% for nanoplastics and 32% for microplastics.\u003c/p\u003e\n\u003cp\u003eOn average, in tonsils, the mass concentrations were 6 times higher for microplastics (\u0026gt;2700 nm) compared to the nanoplastics (20-200 nm). However, particle number concentrations, considering the most conservative sizes for micro- and nanoplastics (2700 and 200 nm respectively), and average concentrations (2500 ng/mg for MPs and 350 ng/mg or NPs), estimate the average particle number of microplastics to ~127013 particles and nanoplastics to ~43750000 particles. In other words, our data shows that, on average, there are 300 times more nanoplastics than microplastic particles in the tonsil samples. This also results in a larger surface area, which can be important for the effects on human health as many reactions occur on surfaces dependent on size and material. For example, the effect on blood coagulation from polystyrene nanoparticles depends on particle surface chemistry\u003csup\u003e54\u003c/sup\u003e and on particle size\u003csup\u003e55\u003c/sup\u003e. However, nanoplastic mechanisms and kinetics around primary uptake, possibly passing into the lymph system or bloodstream, and excretion mechanisms are still mostly theoretical.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNotably, the larger quantities of higher microplastic size fractions (Figures 1a and 2) compared to nanoplastics are consistent with the role of the tonsils as organs that actively sample fine particles for immune response. However, this does not mean that such large plastic particles can effectively enter the lymphatic system and the blood. It is clear that more research is needed to understand the processes that may involve the activation of plastic particles of different sizes and polymer types, as well as their transport kinetics within and between organs in the human body.\u003c/p\u003e\n\u003cp\u003eGiven the limited number of samples, it is not possible to draw conclusions about plastic concentrations at the population level, especially considering the large variation described above. However, it is interesting to note that we observed a weak correlation between body mass index (BMI) and the concentration of PE nanoplastics (size fraction: 20\u0026ndash;200 nm; R = 0.69, p = 0.0047, see SI). Previous studies have suggested that fatty acids and lipid structures may facilitate the retention of plastic particles (PS and PMMA\u0026mdash;polymethyl methacrylate)\u003csup\u003e56\u003c/sup\u003e, but this has not yet been studied in vivo. The relationship between nanoplastic loads and BMI, as well as the mechanisms of retention in fat-rich tissues, would be an important topic for future research with potentially significant toxicological implications.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe want to thank Bengt Olsson, Anna Elfvik, and Natalia Ioukhnenko, and the doctors and the department of ORL surgery, Department of ORL, Ystad Hospital, Ystad, Sweden, for tonsil sample collection, Lena Glantz-Larsson, Department of ORL, Head and Neck Surgery, Sk\u0026aring;ne University Hospital, Lund, Sweden, for logistics support, and Morgan Andersson (deceased), Department of Otorhinolaryngology (ORL), Head and Neck Surgery, Sk\u0026aring;ne University Hospital, Lund, Sweden, for being part of study start-up.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions. \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eM.V., T.C., M.T.E., and DM conceived the project and designed the study. M.V. organised the sample collection and coordinated the project. D.M designed and coordinated the analysis and data collection. R.H provided the instrumentation. V.R.T. performed experiments supervised by M.T.E. and DM. V.R.T and DM performed data analysis and figure preparation. M.V, T.C., and DM provided funding support and/or the resources. DM, V.R.T, M.V. and wrote the original draft. All authors contributed to the manuscript editing.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data needed to evaluate the conclusions in the paper including raw files (measures related to nanoplastics) and processing stages of all the data analysis are available via: DOI:[the link will be available upon the review of the manuscript]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests.\u003c/strong\u003e No competing interests\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eJambeck, J. R.; Walker-Franklin, I. The Impacts of Plastics\u0026rsquo; Life Cycle. \u003cem\u003eOne Earth\u003c/em\u003e \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e6\u003c/em\u003e (6), 600\u0026ndash;606. https://doi.org/10.1016/j.oneear.2023.05.015.\u003c/li\u003e\n\u003cli\u003ePlastics Europe. \u003cem\u003ePlastics - the Facts 2022\u003c/em\u003e. https://plasticseurope.org/knowledge-hub/plastics-the-facts-2022/ (accessed 2023-09-04).\u003c/li\u003e\n\u003cli\u003eAndrady, A. L.; Neal, M. A. Applications and Societal Benefits of Plastics. \u003cem\u003ePhilosophical Transactions of the Royal Society B: Biological Sciences\u003c/em\u003e \u003cstrong\u003e2009\u003c/strong\u003e, \u003cem\u003e364\u003c/em\u003e (1526), 1977\u0026ndash;1984. https://doi.org/10.1098/rstb.2008.0304.\u003c/li\u003e\n\u003cli\u003eEkvall, M. T.; St\u0026aacute;bile, F.; Hansson, L.-A. Nanoplastics Rewire Freshwater Food Webs. \u003cem\u003eCommun Earth Environ\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e5\u003c/em\u003e (1), 1\u0026ndash;7. https://doi.org/10.1038/s43247-024-01646-7.\u003c/li\u003e\n\u003cli\u003eWright, S. L.; Kelly, F. J. Plastic and Human Health: A Micro Issue? \u003cem\u003eEnviron. Sci. Technol.\u003c/em\u003e \u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e51\u003c/em\u003e (12), 6634\u0026ndash;6647. https://doi.org/10.1021/acs.est.7b00423.\u003c/li\u003e\n\u003cli\u003eRevel, M.; Ch\u0026acirc;tel, A.; Mouneyrac, C. Micro(Nano)Plastics: A Threat to Human Health? \u003cem\u003eCurrent Opinion in Environmental Science \u0026amp; Health\u003c/em\u003e \u003cstrong\u003e2018\u003c/strong\u003e, \u003cem\u003e1\u003c/em\u003e, 17\u0026ndash;23. https://doi.org/10.1016/j.coesh.2017.10.003.\u003c/li\u003e\n\u003cli\u003eRamsperger, A. F. R. M.; Bergamaschi, E.; Panizzolo, M.; Fenoglio, I.; Barbero, F.; Peters, R.; Undas, A.; Purker, S.; Giese, B.; Lalyer, C. R.; Tamargo, A.; Moreno-Arribas, M. V.; Grossart, H.-P.; K\u0026uuml;hnel, D.; Dietrich, J.; Paulsen, F.; Afanou, A. K.; Zienolddiny-Narui, S.; Eriksen Hammer, S.; Kringlen Ervik, T.; Graff, P.; Brinchmann, B. C.; Nordby, K.-C.; Wallin, H.; Nassi, M.; Benetti, F.; Zanella, M.; Brehm, J.; Kress, H.; L\u0026ouml;der, M. G. J.; Laforsch, C. Nano- and Microplastics: A Comprehensive Review on Their Exposure Routes, Translocation, and Fate in Humans. \u003cem\u003eNanoImpact\u003c/em\u003e \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e29\u003c/em\u003e, 100441. https://doi.org/10.1016/j.impact.2022.100441.\u003c/li\u003e\n\u003cli\u003eBachmann, M.; Zibunas, C.; Hartmann, J.; Tulus, V.; Suh, S.; Guill\u0026eacute;n-Gos\u0026aacute;lbez, G.; Bardow, A. Towards Circular Plastics within Planetary Boundaries. \u003cem\u003eNat Sustain\u003c/em\u003e \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e6\u003c/em\u003e (5), 599\u0026ndash;610. https://doi.org/10.1038/s41893-022-01054-9.\u003c/li\u003e\n\u003cli\u003eHahladakis, J. N.; Velis, C. A.; Weber, R.; Iacovidou, E.; Purnell, P. An Overview of Chemical Additives Present in Plastics: Migration, Release, Fate and Environmental Impact during Their Use, Disposal and Recycling. \u003cem\u003eJ Hazard Mater\u003c/em\u003e \u003cstrong\u003e2018\u003c/strong\u003e, \u003cem\u003e344\u003c/em\u003e, 179\u0026ndash;199. https://doi.org/10.1016/j.jhazmat.2017.10.014.\u003c/li\u003e\n\u003cli\u003eWiesinger, H.; Wang, Z.; Hellweg, S. Deep Dive into Plastic Monomers, Additives, and Processing Aids. \u003cem\u003eEnviron. Sci. Technol.\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e55\u003c/em\u003e (13), 9339\u0026ndash;9351. https://doi.org/10.1021/acs.est.1c00976.\u003c/li\u003e\n\u003cli\u003eGigault, J.; El Hadri, H.; Nguyen, B.; Grassl, B.; Rowenczyk, L.; Tufenkji, N.; Feng, S.; Wiesner, M. Nanoplastics Are Neither Microplastics nor Engineered Nanoparticles. \u003cem\u003eNat. Nanotechnol.\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e16\u003c/em\u003e (5), 501\u0026ndash;507. https://doi.org/10.1038/s41565-021-00886-4.\u003c/li\u003e\n\u003cli\u003eFrias, J. P. G. L.; Nash, R. Microplastics: Finding a Consensus on the Definition. \u003cem\u003eMarine Pollution Bulletin\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e138\u003c/em\u003e, 145\u0026ndash;147. https://doi.org/10.1016/j.marpolbul.2018.11.022.\u003c/li\u003e\n\u003cli\u003eVethaak, A. D.; Legler, J. Microplastics and Human Health. \u003cem\u003eScience\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e371\u003c/em\u003e (6530), 672\u0026ndash;674. https://doi.org/10.1126/science.abe5041.\u003c/li\u003e\n\u003cli\u003eCasta\u0026ntilde;eda, R. A.; Avlijas, S.; Simard, M. A.; Ricciardi, A. Microplastic Pollution in St. Lawrence River Sediments. \u003cem\u003eCan. J. Fish. Aquat. Sci.\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e71\u003c/em\u003e (12), 1767\u0026ndash;1771. https://doi.org/10.1139/cjfas-2014-0281.\u003c/li\u003e\n\u003cli\u003eDekiff, J. H.; Remy, D.; Klasmeier, J.; Fries, E. Occurrence and Spatial Distribution of Microplastics in Sediments from Norderney. \u003cem\u003eEnvironmental Pollution\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e186\u003c/em\u003e, 248\u0026ndash;256. https://doi.org/10.1016/j.envpol.2013.11.019.\u003c/li\u003e\n\u003cli\u003eKumar, M.; Xiong, X.; He, M.; Tsang, D. C. W.; Gupta, J.; Khan, E.; Harrad, S.; Hou, D.; Ok, Y. S.; Bolan, N. S. Microplastics as Pollutants in Agricultural Soils. \u003cem\u003eEnvironmental Pollution\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e, \u003cem\u003e265\u003c/em\u003e, 114980. https://doi.org/10.1016/j.envpol.2020.114980.\u003c/li\u003e\n\u003cli\u003eIsobe, A.; Uchiyama-Matsumoto, K.; Uchida, K.; Tokai, T. Microplastics in the Southern Ocean. \u003cem\u003eMarine Pollution Bulletin\u003c/em\u003e \u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e114\u003c/em\u003e (1), 623\u0026ndash;626. https://doi.org/10.1016/j.marpolbul.2016.09.037.\u003c/li\u003e\n\u003cli\u003eGalgani, F.; Brien, A. S.; Weis, J.; Ioakeimidis, C.; Schuyler, Q.; Makarenko, I.; Griffiths, H.; Bondareff, J.; Vethaak, D.; Deidun, A.; Sobral, P.; Topouzelis, K.; Vlahos, P.; Lana, F.; Hassellov, M.; Gerigny, O.; Arsonina, B.; Ambulkar, A.; Azzaro, M.; Bebianno, M. J. Are Litter, Plastic and Microplastic Quantities Increasing in the Ocean? \u003cem\u003eMicroplastics and Nanoplastics\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e1\u003c/em\u003e (1), 2. https://doi.org/10.1186/s43591-020-00002-8.\u003c/li\u003e\n\u003cli\u003eHaward, M. Plastic Pollution of the World\u0026rsquo;s Seas and Oceans as a Contemporary Challenge in Ocean Governance. \u003cem\u003eNat Commun\u003c/em\u003e \u003cstrong\u003e2018\u003c/strong\u003e, \u003cem\u003e9\u003c/em\u003e (1), 667. https://doi.org/10.1038/s41467-018-03104-3.\u003c/li\u003e\n\u003cli\u003eMaterić, D.; Holzinger, R.; Niemann, H. Nanoplastics and Ultrafine Microplastic in the Dutch Wadden Sea \u0026ndash; The Hidden Plastics Debris? \u003cem\u003eScience of The Total Environment\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e846\u003c/em\u003e, 157371. https://doi.org/10.1016/j.scitotenv.2022.157371.\u003c/li\u003e\n\u003cli\u003eZhu, X.; Rochman, C. M.; Hardesty, B. D.; Wilcox, C. Plastics in the Deep Sea \u0026ndash; A Global Estimate of the Ocean Floor Reservoir. \u003cem\u003eDeep Sea Research Part I: Oceanographic Research Papers\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e206\u003c/em\u003e, 104266. https://doi.org/10.1016/j.dsr.2024.104266.\u003c/li\u003e\n\u003cli\u003eVan Cauwenberghe, L.; Vanreusel, A.; Mees, J.; Janssen, C. R. Microplastic Pollution in Deep-Sea Sediments. \u003cem\u003eEnvironmental Pollution\u003c/em\u003e \u003cstrong\u003e2013\u003c/strong\u003e, \u003cem\u003e182\u003c/em\u003e, 495\u0026ndash;499. https://doi.org/10.1016/j.envpol.2013.08.013.\u003c/li\u003e\n\u003cli\u003eFinger, J. V. G.; Cor\u0026aacute;, D. H.; Convey, P.; Cruz, F. S.; Petry, M. V.; Kr\u0026uuml;ger, L. Anthropogenic Debris in an Antarctic Specially Protected Area in the Maritime Antarctic. \u003cem\u003eMarine Pollution Bulletin\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e172\u003c/em\u003e, 112921. https://doi.org/10.1016/j.marpolbul.2021.112921.\u003c/li\u003e\n\u003cli\u003eMishra, A. K.; Singh, J.; Mishra, P. P. Microplastics in Polar Regions: An Early Warning to the World\u0026rsquo;s Pristine Ecosystem. \u003cem\u003eScience of The Total Environment\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e784\u003c/em\u003e, 147149. https://doi.org/10.1016/j.scitotenv.2021.147149.\u003c/li\u003e\n\u003cli\u003eKelly, A.; Lannuzel, D.; Rodemann, T.; Meiners, K. M.; Auman, H. J. Microplastic Contamination in East Antarctic Sea Ice. \u003cem\u003eMarine Pollution Bulletin\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e, \u003cem\u003e154\u003c/em\u003e, 111130. https://doi.org/10.1016/j.marpolbul.2020.111130.\u003c/li\u003e\n\u003cli\u003eVelimirovic, M.; Tirez, K.; Voorspoels, S.; Vanhaecke, F. Recent Developments in Mass Spectrometry for the Characterization of Micro- and Nanoscale Plastic Debris in the Environment. \u003cem\u003eAnal Bioanal Chem\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e. https://doi.org/10.1007/s00216-020-02898-w.\u003c/li\u003e\n\u003cli\u003eIvleva, N. P. Chemical Analysis of Microplastics and Nanoplastics: Challenges, Advanced Methods, and Perspectives. \u003cem\u003eChem. Rev.\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e121\u003c/em\u003e (19), 11886\u0026ndash;11936. https://doi.org/10.1021/acs.chemrev.1c00178.\u003c/li\u003e\n\u003cli\u003eJakubowicz, I.; Enebro, J.; Yarahmadi, N. Challenges in the Search for Nanoplastics in the Environment\u0026mdash;A Critical Review from the Polymer Science Perspective. \u003cem\u003ePolymer Testing\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e93\u003c/em\u003e, 106953. https://doi.org/10.1016/j.polymertesting.2020.106953.\u003c/li\u003e\n\u003cli\u003eMaterić, D.; Kj\u0026aelig;r, H. A.; Vallelonga, P.; Tison, J.-L.; R\u0026ouml;ckmann, T.; Holzinger, R. Nanoplastics Measurements in Northern and Southern Polar Ice. \u003cem\u003eEnvironmental Research\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e208\u003c/em\u003e, 112741. https://doi.org/10.1016/j.envres.2022.112741.\u003c/li\u003e\n\u003cli\u003eMaterić, D.; Ludewig, E.; Brunner, D.; R\u0026ouml;ckmann, T.; Holzinger, R. Nanoplastics Transport to the Remote, High-Altitude Alps. \u003cem\u003eEnvironmental Pollution\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, 117697. https://doi.org/10.1016/j.envpol.2021.117697.\u003c/li\u003e\n\u003cli\u003eJurkschat, L.; Gill, A. J.; Milner, R.; Holzinger, R.; Evangeliou, N.; Eckhardt, S.; Materić, D. Using a Citizen Science Approach to Assess Nanoplastics Pollution in Remote High-Altitude Glaciers. \u003cem\u003eSci Rep\u003c/em\u003e \u003cstrong\u003e2025\u003c/strong\u003e, \u003cem\u003e15\u003c/em\u003e (1), 1864. https://doi.org/10.1038/s41598-024-84210-9.\u003c/li\u003e\n\u003cli\u003eAllen, S.; Materić, D.; Allen, D.; MacDonald, A.; Holzinger, R.; Roux, G. L.; Phoenix, V. R. An Early Comparison of Nano to Microplastic Mass in a Remote Catchment\u0026rsquo;s Atmospheric Deposition. \u003cem\u003eJournal of Hazardous Materials Advances\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e7\u003c/em\u003e, 100104. https://doi.org/10.1016/j.hazadv.2022.100104.\u003c/li\u003e\n\u003cli\u003eKirchsteiger, B.; Materić, D.; Happenhofer, F.; Holzinger, R.; Kasper-Giebl, A. Fine Micro- and Nanoplastics Particles (PM2.5) in Urban Air and Their Relation to Polycyclic Aromatic Hydrocarbons. \u003cem\u003eAtmospheric Environment\u003c/em\u003e \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e301\u003c/em\u003e, 119670. https://doi.org/10.1016/j.atmosenv.2023.119670.\u003c/li\u003e\n\u003cli\u003eKau, D.; Materić, D.; Holzinger, R.; Kasper-Giebl, A. \u003cem\u003eFine Microplastics and Nanoplastics in Particulate Matter Samples from a High Alpine Environment\u003c/em\u003e; EGU23-5730; Copernicus Meetings, 2023. https://doi.org/10.5194/egusphere-egu23-5730.\u003c/li\u003e\n\u003cli\u003eV. L. Leonard, S.; Liddle, C. R.; Atherall, C. A.; Chapman, E.; Watkins, M.; D. J. Calaminus, S.; Rotchell, J. M. Microplastics in Human Blood: Polymer Types, Concentrations and Characterisation Using \u0026mu;FTIR. \u003cem\u003eEnvironment International\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e188\u003c/em\u003e, 108751. https://doi.org/10.1016/j.envint.2024.108751.\u003c/li\u003e\n\u003cli\u003eHorvatits, T.; Tamminga, M.; Liu, B.; Sebode, M.; Carambia, A.; Fischer, L.; P\u0026uuml;schel, K.; Huber, S.; Fischer, E. K. Microplastics Detected in Cirrhotic Liver Tissue. \u003cem\u003eeBioMedicine\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e82\u003c/em\u003e. https://doi.org/10.1016/j.ebiom.2022.104147.\u003c/li\u003e\n\u003cli\u003eJenner, L. C.; Rotchell, J. M.; Bennett, R. T.; Cowen, M.; Tentzeris, V.; Sadofsky, L. R. Detection of Microplastics in Human Lung Tissue Using \u0026mu;FTIR Spectroscopy. \u003cem\u003eScience of The Total Environment\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e831\u003c/em\u003e, 154907. https://doi.org/10.1016/j.scitotenv.2022.154907.\u003c/li\u003e\n\u003cli\u003eNihart, A. J.; Garcia, M. A.; El Hayek, E.; Liu, R.; Olewine, M.; Kingston, J. D.; Castillo, E. F.; Gullapalli, R. R.; Howard, T.; Bleske, B.; Scott, J.; Gonzalez-Estrella, J.; Gross, J. M.; Spilde, M.; Adolphi, N. L.; Gallego, D. F.; Jarrell, H. S.; Dvorscak, G.; Zuluaga-Ruiz, M. E.; West, A. B.; Campen, M. J. Bioaccumulation of Microplastics in Decedent Human Brains. \u003cem\u003eNat Med\u003c/em\u003e \u003cstrong\u003e2025\u003c/strong\u003e, 1\u0026ndash;6. https://doi.org/10.1038/s41591-024-03453-1.\u003c/li\u003e\n\u003cli\u003eMontano, L.; Giorgini, E.; Notarstefano, V.; Notari, T.; Ricciardi, M.; Piscopo, M.; Motta, O. Raman Microspectroscopy Evidence of Microplastics in Human Semen. \u003cem\u003eScience of The Total Environment\u003c/em\u003e \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e901\u003c/em\u003e, 165922. https://doi.org/10.1016/j.scitotenv.2023.165922.\u003c/li\u003e\n\u003cli\u003eZhao, Q.; Zhu, L.; Weng, J.; Jin, Z.; Cao, Y.; Jiang, H.; Zhang, Z. Detection and Characterization of Microplastics in the Human Testis and Semen. \u003cem\u003eScience of The Total Environment\u003c/em\u003e \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e877\u003c/em\u003e, 162713. https://doi.org/10.1016/j.scitotenv.2023.162713.\u003c/li\u003e\n\u003cli\u003eHu, C. J.; Garcia, M. A.; Nihart, A.; Liu, R.; Yin, L.; Adolphi, N.; Gallego, D. F.; Kang, H.; Campen, M. J.; Yu, X. Microplastic Presence in Dog and Human Testis and Its Potential Association with Sperm Count and Weights of Testis and Epididymis. \u003cem\u003eToxicological Sciences\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e200\u003c/em\u003e (2), 235\u0026ndash;240. https://doi.org/10.1093/toxsci/kfae060.\u003c/li\u003e\n\u003cli\u003eXu, H.; Dong, C.; Yu, Z.; Hu, Z.; Yu, J.; Ma, D.; Yao, W.; Qi, X.; Ozaki, Y.; Xie, Y. First Identification of Microplastics in Human Uterine Fibroids and Myometrium. \u003cem\u003eEnvironmental Pollution\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e360\u003c/em\u003e, 124632. https://doi.org/10.1016/j.envpol.2024.124632.\u003c/li\u003e\n\u003cli\u003eRagusa, A.; Svelato, A.; Santacroce, C.; Catalano, P.; Notarstefano, V.; Carnevali, O.; Papa, F.; Rongioletti, M. C. A.; Baiocco, F.; Draghi, S.; D\u0026rsquo;Amore, E.; Rinaldo, D.; Matta, M.; Giorgini, E. Plasticenta: First Evidence of Microplastics in Human Placenta. \u003cem\u003eEnvironment International\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e146\u003c/em\u003e, 106274. https://doi.org/10.1016/j.envint.2020.106274.\u003c/li\u003e\n\u003cli\u003eLehner, R.; Weder, C.; Petri-Fink, A.; Rothen-Rutishauser, B. Emergence of Nanoplastic in the Environment and Possible Impact on Human Health. \u003cem\u003eEnviron. Sci. Technol.\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e53\u003c/em\u003e (4), 1748\u0026ndash;1765. https://doi.org/10.1021/acs.est.8b05512.\u003c/li\u003e\n\u003cli\u003eKumar, R.; Manna, C.; Padha, S.; Verma, A.; Sharma, P.; Dhar, A.; Ghosh, A.; Bhattacharya, P. Micro(Nano)Plastics Pollution and Human Health: How Plastics Can Induce Carcinogenesis to Humans? \u003cem\u003eChemosphere\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e298\u003c/em\u003e, 134267. https://doi.org/10.1016/j.chemosphere.2022.134267.\u003c/li\u003e\n\u003cli\u003eZhang, H.; Zhang, S.; Duan, Z.; Wang, L. Pulmonary Toxicology Assessment of Polyethylene Terephthalate Nanoplastic Particles in Vitro. \u003cem\u003eEnvironment International\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e162\u003c/em\u003e, 107177. https://doi.org/10.1016/j.envint.2022.107177.\u003c/li\u003e\n\u003cli\u003eMaterić, D.; Kasper-Giebl, A.; Kau, D.; Anten, M.; Greilinger, M.; Ludewig, E.; van Sebille, E.; R\u0026ouml;ckmann, T.; Holzinger, R. Micro- and Nanoplastics in Alpine Snow: A New Method for Chemical Identification and (Semi)Quantification in the Nanogram Range. \u003cem\u003eEnviron. Sci. Technol.\u003c/em\u003e \u003cstrong\u003e2020\u003c/strong\u003e, \u003cem\u003e54\u003c/em\u003e (4), 2353\u0026ndash;2359. https://doi.org/10.1021/acs.est.9b07540.\u003c/li\u003e\n\u003cli\u003eFraissinet, S.; De Benedetto, G. E.; Malitesta, C.; Holzinger, R.; Materić, D. Microplastics and Nanoplastics Size Distribution in Farmed Mussel Tissues. \u003cem\u003eCommun Earth Environ\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e5\u003c/em\u003e (1), 1\u0026ndash;8. https://doi.org/10.1038/s43247-024-01300-2.\u003c/li\u003e\n\u003cli\u003eCappellin, L.; Karl, T.; Probst, M.; Ismailova, O.; Winkler, P. M.; Soukoulis, C.; Aprea, E.; M\u0026auml;rk, T. D.; Gasperi, F.; Biasioli, F. On Quantitative Determination of Volatile Organic Compound Concentrations Using Proton Transfer Reaction Time-of-Flight Mass Spectrometry. \u003cem\u003eEnviron. Sci. Technol.\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e46\u003c/em\u003e (4), 2283\u0026ndash;2290. https://doi.org/10.1021/es203985t.\u003c/li\u003e\n\u003cli\u003eHolzinger, R.; Acton, W. J. F.; Bloss, W. J.; Breitenlechner, M.; Crilley, L. R.; Dusanter, S.; Gonin, M.; Gros, V.; Keutsch, F. N.; Kiendler-Scharr, A.; Kramer, L. J.; Krechmer, J. E.; Languille, B.; Locoge, N.; Lopez-Hilfiker, F.; Materić, D.; Moreno, S.; Nemitz, E.; Qu\u0026eacute;l\u0026eacute;ver, L. L. J.; Sarda Esteve, R.; Sauvage, S.; Schallhart, S.; Sommariva, R.; Tillmann, R.; Wedel, S.; Worton, D. R.; Xu, K.; Zaytsev, A. Validity and Limitations of Simple Reaction Kinetics to Calculate Concentrations of Organic Compounds from Ion Counts in PTR-MS. \u003cem\u003eAtmospheric Measurement Techniques\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e12\u003c/em\u003e (11), 6193\u0026ndash;6208. https://doi.org/10.5194/amt-12-6193-2019.\u003c/li\u003e\n\u003cli\u003eHansel, A.; Jordan, A.; Holzinger, R.; Prazeller, P.; Vogel, W.; Lindinger, W. Proton Transfer Reaction Mass Spectrometry: On-Line Trace Gas Analysis at the Ppb Level. \u003cem\u003eInternational Journal of Mass Spectrometry and Ion Processes\u003c/em\u003e \u003cstrong\u003e1995\u003c/strong\u003e, \u003cem\u003e149\u0026ndash;150\u003c/em\u003e, 609\u0026ndash;619. https://doi.org/10.1016/0168-1176(95)04294-U.\u003c/li\u003e\n\u003cli\u003eGarcia, M. A.; Liu, R.; Nihart, A.; El Hayek, E.; Castillo, E.; Barrozo, E. R.; Suter, M. A.; Bleske, B.; Scott, J.; Forsythe, K.; Gonzalez-Estrella, J.; Aagaard, K. M.; Campen, M. J. Quantitation and Identification of Microplastics Accumulation in Human Placental Specimens Using Pyrolysis Gas Chromatography Mass Spectrometry. \u003cem\u003eToxicological Sciences\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e199\u003c/em\u003e (1), 81\u0026ndash;88. https://doi.org/10.1093/toxsci/kfae021.\u003c/li\u003e\n\u003cli\u003eMarfella Raffaele; Prattichizzo Francesco; Sardu Celestino; Fulgenzi Gianluca; Graciotti Laura; Spadoni Tatiana; D\u0026rsquo;Onofrio Nunzia; Scisciola Lucia; La Grotta Rosalba; Frig\u0026eacute; Chiara; Pellegrini Valeria; Municin\u0026ograve; Maurizio; Siniscalchi Mario; Spinetti Fabio; Vigliotti Gennaro; Vecchione Carmine; Carrizzo Albino; Accarino Giulio; Squillante Antonio; Spaziano Giuseppe; Mirra Davida; Esposito Renata; Altieri Simona; Falco Giovanni; Fenti Angelo; Galoppo Simona; Canzano Silvana; Sasso Ferdinando C.; Matacchione Giulia; Olivieri Fabiola; Ferraraccio Franca; Panarese Iacopo; Paolisso Pasquale; Barbato Emanuele; Lubritto Carmine; Balestrieri Maria L.; Mauro Ciro; Caballero Augusto E.; Rajagopalan Sanjay; Ceriello Antonio; D\u0026rsquo;Agostino Bruno; Iovino Pasquale; Paolisso Giuseppe. Microplastics and Nanoplastics in Atheromas and Cardiovascular Events. \u003cem\u003eNew England Journal of Medicine\u003c/em\u003e \u003cstrong\u003e2024\u003c/strong\u003e, \u003cem\u003e390\u003c/em\u003e (10), 900\u0026ndash;910. https://doi.org/10.1056/NEJMoa2309822.\u003c/li\u003e\n\u003cli\u003eOslakovic, C.; Cedervall, T.; Linse, S.; Dahlb\u0026auml;ck, B. Polystyrene Nanoparticles Affecting Blood Coagulation. \u003cem\u003eNanomedicine: Nanotechnology, Biology and Medicine\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e8\u003c/em\u003e (6), 981\u0026ndash;986. https://doi.org/10.1016/j.nano.2011.12.001.\u003c/li\u003e\n\u003cli\u003eSanfins, E.; Augustsson, C.; Dahlb\u0026auml;ck, B.; Linse, S.; Cedervall, T. Size-Dependent Effects of Nanoparticles on Enzymes in the Blood Coagulation Cascade. \u003cem\u003eNano Lett.\u003c/em\u003e \u003cstrong\u003e2014\u003c/strong\u003e, \u003cem\u003e14\u003c/em\u003e (8), 4736\u0026ndash;4744. https://doi.org/10.1021/nl501863u.\u003c/li\u003e\n\u003cli\u003eNikpay, M. Polystyrene and Polymethylmethacrylate Microplastics Embedded in Fat, Oil, and Grease (FOG) Deposits of Sewers. \u003cem\u003ePollution\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e8\u003c/em\u003e (4), 1338\u0026ndash;1347. https://doi.org/10.22059/poll.2022.342517.1464.\u003c/li\u003e\n\u003cli\u003eFraissinet, S.; Pennetta, A.; Rossi, S.; De Benedetto, G. E.; Malitesta, C. Optimization of a New Multi-Reagent Procedure for Quantitative Mussel Digestion in Microplastic Analysis. \u003cem\u003eMarine Pollution Bulletin\u003c/em\u003e \u003cstrong\u003e2021\u003c/strong\u003e, \u003cem\u003e173\u003c/em\u003e, 112931. https://doi.org/10.1016/j.marpolbul.2021.112931.\u003c/li\u003e\n\u003cli\u003eSchneider, C. A.; Rasband, W. S.; Eliceiri, K. W. NIH Image to ImageJ: 25 Years of Image Analysis. \u003cem\u003eNat Methods\u003c/em\u003e \u003cstrong\u003e2012\u003c/strong\u003e, \u003cem\u003e9\u003c/em\u003e (7), 671\u0026ndash;675. https://doi.org/10.1038/nmeth.2089.\u003c/li\u003e\n\u003cli\u003eMaterić, D.; Peacock, M.; Kent, M.; Cook, S.; Gauci, V.; R\u0026ouml;ckmann, T.; Holzinger, R. Characterisation of the Semi-Volatile Component of Dissolved Organic Matter by Thermal Desorption \u0026ndash; Proton Transfer Reaction \u0026ndash; Mass Spectrometry. \u003cem\u003eScientific Reports\u003c/em\u003e \u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e7\u003c/em\u003e (1), 15936. https://doi.org/10.1038/s41598-017-16256-x.\u003c/li\u003e\n\u003cli\u003eHolzinger, R. PTRwid: A New Widget Tool for Processing PTR-TOF-MS Data. \u003cem\u003eAtmos. Meas. Tech.\u003c/em\u003e \u003cstrong\u003e2015\u003c/strong\u003e, \u003cem\u003e8\u003c/em\u003e (9), 3903\u0026ndash;3922. https://doi.org/10.5194/amt-8-3903-2015.\u003c/li\u003e\n\u003cli\u003eMaterić, D.; Ludewig, E.; Xu, K.; R\u0026ouml;ckmann, T.; Holzinger, R. Brief Communication: Analysis of Organic Matter in Surface Snow by PTR-MS \u0026ndash; Implications for Dry Deposition Dynamics in the Alps. \u003cem\u003eThe Cryosphere\u003c/em\u003e \u003cstrong\u003e2019\u003c/strong\u003e, \u003cem\u003e13\u003c/em\u003e (1), 297\u0026ndash;307. https://doi.org/10.5194/tc-13-297-2019.\u003c/li\u003e\n\u003cli\u003ePeacock, M.; Materić, D.; Kothawala, D. N.; Holzinger, R.; Futter, M. N. Understanding Dissolved Organic Matter Reactivity and Composition in Lakes and Streams Using Proton-Transfer-Reaction Mass Spectrometry (PTR-MS). \u003cem\u003eEnviron. Sci. Technol. Lett.\u003c/em\u003e \u003cstrong\u003e2018\u003c/strong\u003e, \u003cem\u003e5\u003c/em\u003e (12), 739\u0026ndash;744. https://doi.org/10.1021/acs.estlett.8b00529.\u003c/li\u003e\n\u003cli\u003eYao, P.; Holzinger, R.; Materić, D.; Oyama, B. S.; de F\u0026aacute;tima Andrade, M.; Paul, D.; Ni, H.; Noto, H.; Huang, R.-J.; Dusek, U. Methylsiloxanes from Vehicle Emissions Detected in Aerosol Particles. \u003cem\u003eEnviron. Sci. Technol.\u003c/em\u003e \u003cstrong\u003e2023\u003c/strong\u003e, \u003cem\u003e57\u003c/em\u003e (38), 14269\u0026ndash;14279. https://doi.org/10.1021/acs.est.3c03797.\u003c/li\u003e\n\u003cli\u003eYao, P.; Chianese, E.; Kairys, N.; Holzinger, R.; Materić, D.; Sirignano, C.; Riccio, A.; Ni, H.; Huang, R.-J.; Dusek, U. A Large Contribution of Methylsiloxanes to Particulate Matter from Ship Emissions. \u003cem\u003eEnvironment International\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e165\u003c/em\u003e, 107324. https://doi.org/10.1016/j.envint.2022.107324.\u003c/li\u003e\n\u003cli\u003eMaterić, D.; Peacock, M.; Dean, J.; Futter, M.; Maximov, T.; Moldan, F.; R\u0026ouml;ckmann, T.; Holzinger, R. Presence of Nanoplastics in Rural and Remote Surface Waters. \u003cem\u003eEnviron. Res. Lett.\u003c/em\u003e \u003cstrong\u003e2022\u003c/strong\u003e, \u003cem\u003e17\u003c/em\u003e (5), 054036. https://doi.org/10.1088/1748-9326/ac68f7.\u003c/li\u003e\n\u003cli\u003eEllis, A. M.; Mayhew, C. A. \u003cem\u003eProton Transfer Reaction Mass Spectrometry: Principles and Applications\u003c/em\u003e, 1 edition.; Wiley-Blackwell: Chichester, West Sussex, 2014.\u003c/li\u003e\n\u003cli\u003eThompson, K. M.; Burmaster, D. E.; Crouch3, E. A. C. Monte Carlo Techniques for Quantitative Uncertainty Analysis in Public Health Risk Assessments. \u003cem\u003eRisk Analysis\u003c/em\u003e \u003cstrong\u003e1992\u003c/strong\u003e, \u003cem\u003e12\u003c/em\u003e (1), 53\u0026ndash;63. https://doi.org/10.1111/j.1539-6924.1992.tb01307.x.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-6154338/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6154338/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"This study investigates the presence of micro- and nanoplastics in human tonsil tissue. The tonsils are uniquely positioned in the oropharynx, a gateway to both the immune, respiratory, and digestive systems, thus acting as the first line of defence towards inhaled and ingested particles. We analysed tonsil samples from 15 individuals using Thermal Desorption - Proton Transfer Reaction - Mass Spectrometry, detecting a range of micro- and nanoplastics types, including polystyrene (PS), polyethylene (PE), and polyvinyl chloride (PVC), along with notable findings of tyre wear particles. We detected large differences in polymer types and size classes for each sample with concentrations spanning over four orders of magnitude, bringing nanoplastic concentration for the size class 20-200 nm with an average of 350 ng/mg dry weight. This study is the first to document the accumulation of nanoplastics and nanosized tyre wear in an immunologically active human organ.","manuscriptTitle":"Size distribution of nanoplastics and tyre wear particles in human tonsils","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-31 10:12:06","doi":"10.21203/rs.3.rs-6154338/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"communications-medicine","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"commsmed","sideBox":"Learn more about [Communications Medicine](http://www.nature.com/commsmed)","snPcode":"43856","submissionUrl":"https://mts-commsmed.nature.com/cgi-bin/main.plex","title":"Communications Medicine","twitterHandle":"@commsmedicine","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Communications Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2b0b7824-cd59-498a-8547-36fe0b91dc74","owner":[],"postedDate":"March 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":45585916,"name":"Biological sciences/Chemical biology"},{"id":45585917,"name":"Earth and environmental sciences/Environmental sciences/Environmental chemistry"},{"id":45585918,"name":"Physical sciences/Chemistry/Chemical biology"},{"id":45585919,"name":"Biological sciences/Biological techniques"}],"tags":[],"updatedAt":"2025-03-31T10:12:06+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-31 10:12:06","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6154338","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6154338","identity":"rs-6154338","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}