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

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

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

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

Municipal biowaste treatment plants contribute to the contamination of the environment with residues of biodegradable plastics with putative higher persistence potential.

Biodegradable plastics (BDP) are expected to mineralize easily, in particular under conditions of technical composting. However, the complexity of the sample matrix has largely prevented degradation studies under realistic conditions. Here composts and fertilizers from state-of-the-art municipal combined anaerobic/aerobic biowaste treatment plants were investigated for residues of BDP. We found BDP fragments > 1 mm in significant numbers in the final composts intended as fertilizer for agriculture and gardening. Compared to pristine compostable bags, the recovered BDP fragments showed differences in their material properties, which potentially renders them less prone to further biodegradation. BDP fragments < 1 mm were extracted in bulk and came up to 0.43 wt% of compost dry weight. Finally, the liquid fertilizer produced during the anaerobic treatment contained several thousand BDP fragments < 500 µm per liter. Hence, our study questions, if currently available BDP are compatible with applications in areas of environmental relevance, such as fertilizer production.

Flooding frequency and floodplain topography determine abundance of microplastics in an alluvial Rhine soil.

Rivers are major pathways for the transport of microplastics towards the oceans, and many studies focus on microplastic abundance in fluvial ecosystems. Although flooding strongly affects transport of microplastics, knowledge about the potential input via floodwaters, spatial distribution, and fate of microplastics in adjacent floodplains remains very limited. In this study, we suggest that local topography and flood frequency could influence the abundance of microplastics in floodplains. Based on this concept, we took soil samples in a Rhine River floodplain in two different depths (0-5 cm and 5-20 cm) along three transects with increasing distance to the river and analysed the abundance of microplastics via FTIR spectroscopy. Flood frequency of the transects was estimated by a combination of hydrodynamic modelling with MIKE 21 (DHI, Hørsholm Denmark) and analysis of time series of water levels. Microplastic abundance per kg dry soil varied between 25,502 to 51,119 particles in the top 5 cm and 25,616 to 84,824 particles in the deeper soil (5-20 cm). The results of our study indicate that local topography and resulting flooding patterns are responsible for the amount of microplastics found at the respective transect. Differences in soil properties, vegetation cover and signs of earthworm activity in the soil profile seem to be related to microplastic migration and accumulation in the deeper soil. The interdisciplinary approach we used in our work can be applied to other floodplains to elucidate the respective processes. This information is essentially important both for locating potential microplastic sinks for process-informed sampling designs and to identify areas of increased bioavailability of microplastics for proper ecological risk assessment.

Microplastic sample purification methods - Assessing detrimental effects of purification procedures on specific plastic types.

In search of effective, fast, and cheap methods to purify environmental samples for microplastic analysis, scientific literature provides various purification protocols. However, while most of these protocols effectively purify the samples, some may also degrade the targeted polymers. This study was conducted to systematically compare the effects of purification protocols based on acidic, alkaline, oxidative, and enzymatic digestion and extraction via density separation on eight of the most relevant plastic types. It offers insights into how specific purification protocols may compromise microplastic detection by documenting visible and gravimetric effects, analyzing potential surface degradation using Fourier transform infrared spectroscopy (FTIR) and bulk erosion on a molecular level using gel permeation chromatography (GPC). For example, protocols using strong acids and high temperatures are likely to completely dissolve or cause strong degradation to a wide range of polymers (PA, PC, PET, PS, PUR & PVC), while strong alkaline solutions may damage PC and PET. Contrarily, Fenton's reagent, multiple enzymatic digestion steps, as well as treatment with a zinc chloride solution frequently used for density-separation, do not degrade the eight polymers tested here. Therefore, their implementation in microplastic sample processing may be considered an essential stepping-stone towards a standardized protocol for future microplastics analyses.

Computer-Assisted Analysis of Microplastics in Environmental Samples Based on µFTIR Imaging in Combination with Machine Learning.

The problem of automating the data analysis of microplastics following a spectroscopic measurement such as focal plane array (FPA)-based micro-Fourier transform infrared (FTIR), Raman, or QCL is gaining ever more attention. Ease of use of the analysis software, reduction of expert time, analysis speed, and accuracy of the result are key for making the overall process scalable and thus allowing nonresearch laboratories to offer microplastics analysis as a service. Over the recent years, the prevailing approach has been to use spectral library search to automatically identify spectra of the sample. Recent studies, however, showed that this approach is rather limited in certain contexts, which led to developments for making library searches more robust but on the other hand also paved the way for introducing more advanced machine learning approaches. This study describes a model-based machine learning approach based on random decision forests for the analysis of large FPA-µFTIR data sets of environmental samples. The model can distinguish between more than 20 different polymer types and is applicable to complex matrices. The performance of the model under these demanding circumstances is shown based on eight different data sets. Further, a Monte Carlo cross validation has been performed to compute error rates such as sensitivity, specificity, and precision.

Tackling the Challenge of Extracting Microplastics from Soils: A Protocol to Purify Soil Samples for Spectroscopic Analysis.

Microplastic pollution in soils is an emerging topic in the scientific community, with researchers striving to determine the occurrence and the impact of microplastics on soil health, ecology, and functionality. However, information on the microplastic contamination of soils is limited because of a lack of suitable analytical methods. Because micro-Fourier-transform infrared spectroscopy (µ-FTIR), next to Raman spectroscopy, is one of the few methods that allows the determination of the number, polymer type, shape, and size of microplastic particles, the present study addresses the challenge of purifying soil samples sufficiently to allow a subsequent µ-FTIR analysis. A combination of freeze-drying, sieving, density separation, and a sequential enzymatic-oxidative digestion protocol enables removal of the mineral mass (>99.9% dry wt) and an average reduction of 77% dry weight of the remaining organic fraction. In addition to visual integrity, attenuated total reflectance FTIR, gel permeation chromatography, and differential scanning calorimetry showed that polyamide, polyethylene, polyethylene terephthalate, and polyvinyl chloride in the size range of 100 to 400 µm were not affected by the approach. However, biodegradable polylactic acid showed visible signs of degradation and reduced molecular weight distribution after protease treatment. Nevertheless, the presented purification protocol is a reliable and robust method to purify relatively large soil samples of approximately 250 g dry weight for spectroscopic analysis in microplastic research and has been shown to recover various microplastic fibers and fragments down to a size of 10 µm from natural soil samples. Environ Toxicol Chem 2022;41:844-857. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Finding Microplastics in Soils: A Review of Analytical Methods.

Research on microplastics in soils is still uncommon, and the existing publications are often incomparable due to the use of different sampling, processing, and analytical methods. Given the complex nature of soils, a suitable and efficient method for standardized microplastic analysis in the soil matrix has yet to be found. This paper proposes a critical review on the different published methods for sampling, extraction, purification, and identification/quantification of microplastics in complex environmental matrices, with the main focus on their applicability for soil samples. While large microplastic particles can be manually sorted out and verified with chemical analysis, sample preparation for smaller microplastic analysis is usually more difficult. Of the analytical approaches proposed in the literature, some are established, whereas others are a proof of principle and have not yet been applied to environmental samples. For the sake of development, all approaches are discussed and assessed for their potential applicability for soil samples. So far, none of the published methods seems ideally suitable for the analysis of smaller microplastics in soil samples, but slight modifications and combinations of methods may prove promising and need to be explored.

Organic fertilizer as a vehicle for the entry of microplastic into the environment.

The contamination of the environment with microplastic, defined as particles smaller than 5 mm, has emerged as a global challenge because it may pose risks to biota and public health. Current research focuses predominantly on aquatic systems, whereas comparatively little is known regarding the sources, pathways, and possible accumulation of plastic particles in terrestrial ecosystems. We investigated the potential of organic fertilizers from biowaste fermentation and composting as an entry path for microplastic particles into the environment. Particles were classified by size and identified by attenuated total reflection-Fourier transform infrared spectroscopy. All fertilizer samples from plants converting biowaste contained plastic particles, but amounts differed significantly with substrate pretreatment, plant, and waste (for example, household versus commerce) type. In contrast, digestates from agricultural energy crop digesters tested for comparison contained only isolated particles, if any. Among the most abundant synthetic polymers observed were those used for common consumer products. Our results indicate that depending on pretreatment, organic fertilizers from biowaste fermentation and composting, as applied in agriculture and gardening worldwide, are a neglected source of microplastic in the environment.