The Umbilical Cord Creatine Flux and Time Course of Human Milk Creatine across Lactation.

(1) Background: The aim of the present paper was to study fetal and infant creatine (Cr) supply to improve nutrition and neuroprotection in term and especially in preterm infants. The primary outcomes were the placental Cr flux at the end of pregnancy and the time course of human milk (HM) Cr. (2) Methods: The estimation of placental Cr flux was based on umbilical arterial and venous cord blood Cr in 10 term infants after elective caesarian section. HM Cr, creatinine (Crn), and macronutrients were measured longitudinally in 10 mothers across the first 6 months of breastfeeding. (3) Results: At the end of pregnancy, the mean fetal Cr flux was negative (-2.07 mmol/min). HM Cr was highest in colostrum, decreased significantly within the first 2 weeks of breastfeeding (p < 0.05), and did not change significantly thereafter. HM Cr was not correlated with HM Crn or macronutrient composition. (4) Conclusions: The present data suggest that fetal endogenous Cr synthesis covers the needs at the end of pregnancy. However, high colostrum Cr and HM Cr levels, independent of macronutrient composition, suggest that there may be a critical Cr demand immediately after birth that needs to be covered by enteral supply.

In-ovo feeding with creatine monohydrate: implications for chicken energy reserves and breast muscle development during the pre-post hatching period.

The most dynamic period throughout the lifespan of broiler chickens is the pre-post-hatching period, entailing profound effects on their energy status, survival rate, body weight, and muscle growth. Given the significance of this pivotal period, we evaluated the effect of in-ovo feeding (IOF) with creatine monohydrate on late-term embryos' and hatchlings' energy reserves and post-hatch breast muscle development. The results demonstrate that IOF with creatine elevates the levels of high-energy-value molecules (creatine and glycogen) in the liver, breast muscle and yolk sac tissues 48 h post IOF, on embryonic day 19 (p < 0.03). Despite this evidence, using a novel automated image analysis tool on day 14 post-hatch, we found a significantly higher number of myofibers with lower diameter and area in the IOF creatine group compared to the control and IOF NaCl groups (p < 0.004). Gene expression analysis, at hatch, revealed that IOF creatine group had significantly higher expression levels of myogenin (MYOG) and insulin-like growth factor 1 (IGF1), related to differentiation of myogenic cells (p < 0.01), and lower expression of myogenic differentiation protein 1 (MyoD), related to their proliferation (p < 0.04). These results imply a possible effect of IOF with creatine on breast muscle development through differential expression of genes involved in myogenic proliferation and differentiation. The findings provide valuable insights into the potential of pre-hatch enrichment with creatine in modulating post-hatch muscle growth and development.

Supply and demand of creatine and glycogen in broiler chicken embryos.

Optimal embryonic development and growth of meat-type chickens (broilers) rely on incubation conditions (oxygen, heat, and humidity), on nutrients and on energy resources within the egg. Throughout incubation and according to the embryo's energy balance, the main energy storage molecules (creatine and glycogen) are continuously utilized and synthesized, mainly in the embryonic liver, breast muscle, and the extraembryonic yolk sac (YS) tissue. During the last phase of incubation, as the embryo nears hatching, dynamic changes in energy metabolism occur. These changes may affect embryonic survival, hatchlings' uniformity, quality and post hatch performance of broilers, hence, being of great importance to poultry production. Here, we followed the dynamics of creatine and glycogen from embryonic day (E) 11 until hatch and up to chick placement at the farm. We showed that creatine is stored mainly in the breast muscle while glycogen is stored mainly in the YS tissue. Analysis of creatine synthesis genes revealed their expression in the liver, kidney, YS tissue and in the breast muscle, suggesting a full synthesis capacity in these tissues. Expression analysis of genes involved in gluconeogenesis, glycogenesis, and glycogenolysis, revealed that glycogen metabolism is most active in the liver. Nevertheless, due to the relatively large size of the breast muscle and YS tissue, their contribution to glycogen metabolism in embryos is valuable. Towards hatch, post E19, creatine levels in all tissues increased while glycogen levels dramatically decreased and reached low levels at hatch and at chick placement. This proves the utmost importance of creatine in energy supply to late-term embryos and hatchlings.

Addition of a combination of creatine, carnitine, and choline to a commercial diet increases postprandial plasma creatine and creatinine concentrations in adult dogs.

Creatine is a nitrogenous compound essential for cellular energy homeostasis found in animal protein; however, when heat-processed for pet food, creatine is degraded to creatinine, which is not metabolically active and excreted in urine. The objective of the present investigation was to define the postprandial plasma creatine and creatinine response in dogs fed a commercial diet (CON) formulated for adult dogs, top-dressed with a combination of creatine (9.6 g/kg dry matter, DM), carnitine (2.13 g/kg DM) and choline (0.24 g/kg DM; CCC), methionine (2.6 g/kg DM; MET), or taurine (0.7 g/kg DM; TAU). Eight adult Beagles were fed one of the four diets for 7 days in a Latin Square design with no washout period. On day 7, cephalic catheters were placed and blood samples were collected before being fed (fasted) and up to 6 h post-meal. Creatine and creatinine were analyzed using HPLC and data analyzed using PROC GLIMMIX in SAS. Plasma creatine concentrations were higher in dogs fed CCC (103 ± 10 µmol/L) compared to MET (72 ± 7 µmol/L) at fasted (P < 0.05) and higher compared to all other treatments from 15 to 360 min post-meal (P < 0.05). Plasma creatinine concentrations were higher in dogs fed CCC from 60 to 180 min compared to all other treatments. These data suggest that when creatine, carnitine and choline are top-dressed for 7 days, plasma creatine is rapidly absorbed and remains elevated up to 6 h post-meal. This may have implications for energy metabolism and should be considered when using creatinine as a diagnostic tool in dogs.

Two types of microplastics (polystyrene-HBCD and car tire abrasion) affect oxidative stress-related biomarkers in earthworm Eisenia andrei in a time-dependent manner.

Microplastics are small plastic fragments that are widely distributed in marine and terrestrial environments. While the soil ecosystem represents a large reservoir for plastic, research so far has focused mainly on the impact on aquatic ecosystems and there is a lack of information on the potentially adverse effects of microplastics on soil biota. Earthworms are key organisms of the soil ecosystem and are due to their crucial role in soil quality and fertility a suitable and popular model organism in soil ecotoxicology. Therefore, the aim of this study was to gain insight into the effects of environmentally relevant concentrations of microplastics on the earthworm Eisenia andrei on multiple levels of biological organization after different exposure periods. Earthworms were exposed to two types of microplastics: (1) polystyrene-HBCD and (2) car tire abrasion in natural soil for 2, 7, 14 and 28d. Acute and chronic toxicity and all subcellular investigations were conducted for all exposure times, avoidance behavior assessed after 48 h and reproduction after 28d. Subcellular endpoints included enzymatic biomarker responses, namely, carboxylesterase, glutathione peroxidase, acetylcholinesterase, glutathione reductase, glutathione S-transferase and catalase activities, as well as fluorescence-based measurements of oxidative stress-related markers and multixenobiotic resistance activity. Multiple biomarkers showed significant changes in activity, but a recovery of most enzymatic activities could be observed after 28d. Overall, only minor effects could be observed on a subcellular level, showing that in this exposure scenario with environmentally relevant concentrations based on German pollution levels the threat to soil biota is minimal. However, in areas with higher concentrations of microplastics in the environment, these results can be interpreted as an early warning signal for more adverse effects. In conclusion, these findings provide new insights regarding the ecotoxicological effects of environmentally relevant concentrations of microplastics on soil organisms.

Determination of tire wear markers in soil samples and their distribution in a roadside soil.

Tire wear (TW) constitutes a significant source of microplastic in terrestrial ecosystems. It is known that particles emitted by roads can have an effect up to 100 m into adjacent areas. Here, we apply for the first-time thermal extraction desorption gas chromatography-mass spectrometry (TED-GC/MS) to determine TW in soil samples by detection of thermal decomposition products of styrene-butadiene rubber (SBR), without additional enrichment. Additionally, zinc contents were determined as an elemental marker for TW. Mixed soil samples were taken along three transects along a German motorway in 0.3, 2.0, and 5.0 m distance from the road. Sampling depths were 0-2, 2-5, 5-10, and 10-20 cm. Four fine fractions, 1 000-500, 500-100, 100-50, and <50 µm, were analyzed. TW contents based on SBR ranged from 155 to 15 898 mg kg-1. TW contents based on zinc were between 413 and 44 812 mg kg-1. Comparison of individual values of SBR and zinc reveals SBR as a more specific marker. Results confirm that most TW ends up in the topsoil within a 2 m distance. The sampling strategy resulted in representative data for a larger area. Standard deviations of quadruple TED-GC/MS determination of SBR were <10% for all grain size fractions. TED-GC/MS is a suitable analytical tool for determining TW in soil samples without the use of toxic chemicals, enrichment, or special sample preparation.

PhenoApp: A mobile tool for plant phenotyping to record field and greenhouse observations.

With the ongoing cost decrease of genotyping and sequencing technologies, accurate and fast phenotyping remains the bottleneck in the utilizing of plant genetic resources for breeding and breeding research. Although cost-efficient high-throughput phenotyping platforms are emerging for specific traits and/or species, manual phenotyping is still widely used and is a time- and money-consuming step. Approaches that improve data recording, processing or handling are pivotal steps towards the efficient use of genetic resources and are demanded by the research community. Therefore, we developed PhenoApp, an open-source Android app for tablets and smartphones to facilitate the digital recording of phenotypical data in the field and in greenhouses. It is a versatile tool that offers the possibility to fully customize the descriptors/scales for any possible scenario, also in accordance with international information standards such as MIAPPE (Minimum Information About a Plant Phenotyping Experiment) and FAIR (Findable, Accessible, Interoperable, and Reusable) data principles. Furthermore, PhenoApp enables the use of pre-integrated ready-to-use BBCH (Biologische Bundesanstalt für Land- und Forstwirtschaft, Bundessortenamt und CHemische Industrie) scales for apple, cereals, grapevine, maize, potato, rapeseed and rice. Additional BBCH scales can easily be added. The simple and adaptable structure of input and output files enables an easy data handling by either spreadsheet software or even the integration in the workflow of laboratory information management systems (LIMS). PhenoApp is therefore a decisive contribution to increase efficiency of digital data acquisition in genebank management but also contributes to breeding and breeding research by accelerating the labour intensive and time-consuming acquisition of phenotyping data.

The physiological role of guanidinoacetic acid and its relationship with arginine in broiler chickens.

The role of guanidinoacetic acid (GAA) and its relationship with arginine was reviewed in order to define a replacement ratio between GAA and arginine for broiler diet formulation, the ratio being of how much arginine could be spared, or replaced by GAA. Guanidionoacetic acid, the precursor of creatine, can be synthesized de novo from the amino acids arginine and glycine, whereby 1 mol of arginine creates 1 mol of GAA; that is a weight:weight (w:w) ratio of 1.49:1 (arginine:GAA). Guanidinoacetic acid exerts a growth effect through its primary physiological fate to form creatine, and additionally spares dietary arginine from GAA synthesis; so that it contributes to protein accretion and other functions. Creatine is critical in energy metabolism as a carrier and reservoir of phosphate for adenosine triphosphate (ATP) formation. Arginine deficiency causes reduced growth and can lead to disrupted levels of blood and muscle energy metabolites (phosphocreatine and creatine). Supplementing GAA into the diet restores these metabolites. At severe arginine deficiency, GAA addition cannot fully compensate the arginine deficit, as measured by growth performance. As arginine becomes nearer to sufficiency, the effect of GAA becomes more pronounced. When using growth rate or FCR as an indicator in broilers, a ratio in the range of 0.77 to 1.3:1 (w:w arginine:GAA) was seen, with one study noting a ratio of 2:1 when using FCR as an indicator. Higher ratios of up to 2.7:1 are achieved when using muscle creatine and phosphocreatine measurements. A recommendation of 1:1 (w:w) is proposed, which takes a conservative approach. Large scale studies with practical diets would be helpful to confirm that a ratio of 1:1 (w:w) or higher may be used in the field for broilers.

Smart filters for the analysis of microplastic in beverages filled in plastic bottles.

The occurrence of microplastic (MP) in food products, such as beverages in plastic bottles, is of high public concern. Existing analytical methods focus on the determination of particle numbers, requiring elaborate sampling tools, laboratory infrastructure and generally time-consuming imaging detection methods. A comprehensive routine analysis of MP in food products is still not possible. In the present work, we present the development of a smart filter crucible as sampling and detection tool. After filtration and drying of the filtered-off solids, a direct determination of the MP mass content from the crucible sample can be done by thermal extraction desorption gas chromatography mass spectroscopy (TED-GC/MS). The new filter crucible allows a filtration of MP down to particle sizes of 5 µm. We determined MP contents below 0.01 µg/L up to 2 µg/L, depending on beverages bottle type. This may be directly related to the bottle type, especially the quality of the plastic material of the screw cap. Dependent on the plastic material, particle formation increases due to opening and closing operations during the use phase. However, we have also found that some individual determinations of samples were subjected to high errors due to random events. A conclusive quantitative evaluation of the products is therefore not possible at present.

Comprehensive Characterization of APTES Surface Modifications of Hydrous Boehmite Nanoparticles.

Hydrous boehmite (γ-AlOOH) nanoparticles (BNP) show great potential as nanoscale filler for the fabrication of fiber reinforced nanocomposite materials. Notably, the particle-matrix interaction has been demonstrated to be decisive for improving the matrix-dominant mechanical properties in the past years. Tailoring the surface properties of the nanofiller enables to selectively design the interaction and thus to exploit the benefits of the nanocomposite in an optimal way. Here, an extensive study is presented on the binding of (3-aminopropyl)triethoxysilane (APTES), a common silane surface modifier, on BNP in correlation to different process parameters (concentration, time, temperature, and pH). Furthermore, a comprehensive characterization of the modified BNP was performed by using elemental analysis (EA), thermogravimetric analysis (TGA) coupled with mass spectrometry (TGA-MS), and Kaiser's test (KT). The results show an increasing monolayer formation up to a complete surface coverage with rising APTES concentration, time, and temperature, resulting in a maximal grafting density of 1.3 molecules/nm2. Unspecific multilayer formation was solely observed under acidic conditions. Comparison of TGA-MS results with data recorded from EA, TGA, and KT verified that TGA-MS is a convenient and highly suitable method to elucidate the ligand binding in detail.

Color Intensity of the Red-Fleshed Berry Phenotype of Vitis vinifera Teinturier Grapes Varies Due to a 408 bp Duplication in the Promoter of VvmybA1.

Grapevine (Vitis vinifera) teinturier cultivars are characterized by their typical reddish leaves and red-fleshed berries due to ectopic anthocyanin formation. Wines of these varieties have economic importance as they can be used for blending to enhance the color of red wines. The unique and heritable mutation has been known for a long time but the underlying genetic mechanism still is not yet understood. Here we describe the association of the red-fleshed berry phenotype with a 408 bp repetitive DNA element in the promoter of the VvmybA1 gene (grapevine color enhancer, GCE). Three different clones of 'Teinturier' were discovered with two, three and five allelic GCE repeats (MybA1t2, MybA1t3 and MybA1t5). All three clones are periclinal chimeras; these clones share the same L1 layer, but have distinct L2 layers with different quantities of GCE repeats. Quantitative real time PCR and HPLC analysis of leaf and berry samples showed that the GCE repeat number strongly correlates with an increase of the expression of VvmybA1 itself and the VvUFGT gene regulated by it and the anthocyanin content. A model is proposed based on autoregulation of VvmybA1t to explain the red phenotype which is similar to that of red-fleshed apples. This study presents results about the generation and modes of action of three MybA1t alleles responsible for the red-fleshed berry phenotype of teinturier grapevines.

Characterization of tire and road wear particles from road runoff indicates highly dynamic particle properties.

Tire and road wear particles (TRWPs) are heteroagglomerates of tire rubber and other particles deposited on the road surface and one of the main contributors to non-exhaust emissions of automobile traffic. In this study, samples from road environments were analyzed for their TRWP contents and concentrations of eight organic tire constituents. TRWP concentrations were determined by quantifying Zn in the density fraction <1.9 g/cm³ and by thermal extraction desorption-gas chromatography-mass spectrometry (TED-GC/MS) and the concentrations ranged from 3.7 to 480 mg TRWP/g. Strong and statistically significant correlations with TRWPs were found for 2-hydroxybenzothiazole and 2-aminobenzothiazole, indicating that these substances may be suitable markers of TRWPs. The mass distribution of TRWPs in road dust suggests that the main mass fraction formed on roads consists of coarse particles (>100 µm). Data for a sedimentation basin indicate that the fine fraction (<50 µm) is preferentially transported by road runoff into receiving waters. The size distribution and density data of TRWP gathered by three different quantitation approaches also suggest that aging of TRWPs leads to changes in their particle density. An improved understanding of the dynamics of TRWP properties is essential to assess the distribution and dissipation of this contaminant of emerging concern in the environment.

From broiler breeder hen feed to the egg and embryo: The molecular effects of guanidinoacetate supplementation on creatine transport and synthesis.

Supplementation of broiler breeder hens with beneficial additives bears great potential for affecting nutrient deposition into the fertile egg. Guanidinoacetate (GAA) is the endogenous precursor of creatine that is used as a feed additive for improving cellular energy metabolism in animal nutrition. In the present study, we have investigated whether GAA supplementation in broiler breeder feed affects creatine deposition into the hatching egg and molecular mechanisms of creatine transport and synthesis within hens and their progeny. For this, broiler breeder hens of 47 wk of age were supplemented with 0.15% GAA for 15 wk, and samples from their tissues, hatching eggs and progeny were compared with those of control, nonsupplemented hens. A significant increase in creatine content was found within the yolk and albumen of hatching eggs obtained from the GAA group, compared with the control group. The GAA group exhibited a significant increased creatine transporter gene expression compared with the control group in their small intestines and oviduct. In GAA group progeny, a significant decrease in creatine transporter expression at embryonic day 19 and day of hatch was found, compared with control group progeny. At the day of hatch, creatine synthesis genes (arginine glycine amidinotransferase and guanidinoacetate N-methyltransferase) exhibited significant decrease in expression in the GAA group progeny compared with control group progeny. These results indicate that GAA supplementation in broiler breeder feed increases its absorbance and deposition into hatching eggs, subsequently affecting GAA and creatine absorbance and synthesis within broiler progeny.

Specific adsorption sites and conditions derived by thermal decomposition of activated carbons and adsorbed carbamazepine.

The adsorption of organic micropollutants onto activated carbon is a favourable solution for the treatment of drinking water and wastewater. However, these adsorption processes are not sufficiently understood to allow for the appropriate prediction of removal processes. In this study, thermogravimetric analysis, alongside evolved gas analysis, is proposed for the characterisation of micropollutants adsorbed on activated carbon. Varying amounts of carbamazepine were adsorbed onto three different activated carbons, which were subsequently dried, and their thermal decomposition mechanisms examined. The discovery of 55 different pyrolysis products allowed differentiations to be made between specific adsorption sites and conditions. However, the same adsorption mechanisms were found for all samples, which were enhanced by inorganic constituents and oxygen containing surface groups. Furthermore, increasing the loadings led to the evolution of more hydrated decomposition products, whilst parts of the carbamazepine molecules were also integrated into the carbon structure. It was also found that the chemical composition, especially the degree of dehydration of the activated carbon, plays an important role in the adsorption of carbamazepine. Hence, it is thought that the adsorption sites may have a higher adsorption energy for specific adsorbates, when the activated carbon can then potentially increase its degree of graphitisation.

Effects of Defoliation on Phenolic Concentrations, Antioxidant and Antibacterial Activity of Grape Skin Extracts of the Varieties Blaufränkisch and Merlot (Vitis vinifera L.).

Vitis vinifera L. has been highlighted by its many positive effects on human health, including antibacterial, anti-inflammatory and antioxidant activity due to its rich phytochemical content. Removing six basal leaves has great potential to influence the production of phytochemicals. The purpose of this study was to determine the impact of different terms of defoliation on the antioxidant and antibacterial activity of grape skin extracts of the Blaufränkisch and Merlot varieties. The total phenolic concentration, total and individual anthocyanin concentration, antioxidant activity and antibacterial activity on gram-positive and gram-negative human pathogens have been determined. The study was conducted on three treatments (control treatment without defoliation, defoliation immediately after bloom and defoliation before the start of the véraison phase). The results showed statistically significant enhancement of the total phenolic concentration as well as the antioxidant and antibacterial activity in both studied cultivars. Defoliation just after blooming was the preferable defoliation term in the Merlot variety for achieving the highest total anthocyanin concentration, antioxidant activity and significant increase of antibacterial activity against all four investigated bacteria. Defoliation before the start of the véraison phase was the preferable defoliation term for achieving the highest total anthocyanin concentration in the Blaufränkisch variety. In general, treatment of defoliation immediately after bloom was more beneficial compared with the defoliation before the start of the véraison phase.

Stability of creatine monohydrate and guanidinoacetic acid during manufacture (retorting and extrusion) and storage of dog foods.

The stability of creatine monohydrate (CrMH), crystallised guanidinoacetic acid (GAA-C) and granulated GAA (GAA-G) in a moist retorted and a dry extruded dog food formulation during production and storage was investigated. Commercial food mixtures were supplemented with CrMH, GAA-C or GAA-G. Uniformity after mixing and retorting or extrusion was determined based on replicate samples (moist n = 8, dry n = 10). Storage stability was evaluated at 25°C/60% relative humidity for 15 months and 40°C/75% for 6 months. Foods with CrMH were analysed for creatine (Cr) and creatinine (Crn), whereas GAA-C and GAA-G foods were analysed for GAA concentrations. Coefficients of variation (CV) for uniformity of the additives after mixing of moist and dry pet food formulations were below 15%, and the CV was lower in processed mixtures. Recoveries after retorting and extrusion were higher for GAA-G (79 and 99%) and GAA-C (89 and 86%) compared to CrMH (36 and 85%) foods. In moist CrMH food, Cr concentrations re-increased by 54% whilst Crn concentrations decreased by 39% after storage at 25°C for 15 months. With total molar Cr + Crn remaining stable throughout storage, Crn and Cr appeared to effectively interconvert. Storage of the extruded CrMH food at 25°C for 15 months resulted in a 63% decrease in Cr and a 39% increase in Crn concentration. The decrease in Cr concentration was larger at 6 months storage at 40°C compared to 15 months storage at 25°C. Both GAA-C and GAA-G moist and dry foods were stable during storage (<10% decrease). This study showed that GAA is highly stable during production and storage of moist and dry canine foods whilst CrMH is relatively unstable, particularly during storage. The latter makes it difficult to establish a guaranteed Cr content in finished moist retorted and dry extruded foods with CrMH.

Boehmite Nanofillers in Epoxy Oligosiloxane Resins: Influencing the Curing Process by Complex Physical and Chemical Interactions.

In this work, a novel boehmite (BA)-embedded organic/inorganic nanocomposite coating based on cycloaliphatic epoxy oligosiloxane (CEOS) resin was fabricated applying UV-induced cationic polymerization. The main changes of the material behavior caused by the nanofiller were investigated with regard to its photocuring kinetics, thermal stability, and glass transition. The role of the particle surface was of particular interest, thus, unmodified nanoparticles (HP14) and particles modified with p-toluenesulfonic acid (OS1) were incorporated into a CEOS matrix in the concentration range of 1-10 wt.%. Resulting nanocomposites exhibited improved thermal properties, with the glass transition temperature (Tg) being shifted from 30 °C for unfilled CEOS to 54 °C (2 wt.% HP14) and 73 °C (2 wt.% OS1) for filled CEOS. Additionally, TGA analysis showed increased thermal stability of samples filled with nanoparticles. An attractive interaction between boehmite and CEOS matrix influenced the curing. Real-time infrared spectroscopy (RT-IR) experiments demonstrated that the epoxide conversion rate of nanocomposites was slightly increased compared to neat resin. The beneficial role of the BA can be explained by the participation of hydroxyl groups at the particle surface in photopolymerization processes and by the complementary contribution of p-toluenesulfonic acid surface modifier and water molecules introduced into the system with nanoparticles.

Tire and road wear particles in road environment - Quantification and assessment of particle dynamics by Zn determination after density separation.

In this study, a method for the determination of tire and road wear particle (TRWP) contents in particulate samples from road environment was developed. Zn was identified as the most suitable elemental marker for TRWP, due to its high concentration in tire tread and the possibility of separation from other Zn sources. The mean concentration of 21 tire samples was 8.7 ± 2.0 mg Zn/g. Before quantification in samples from road environment, TRWP were separated from the particulate matrix by density separation. Method development was conducted using shredded tread particles (TP) as a surrogate for TRWP. Recovery of TP from spiked sediment was 95 ± 17% in a concentration range of 2 - 200 mg TP/g. TP determination was not affected by other Zn containing solids or spiked Zn-salts. By adjusting the density of the separation solution to 1.9 g/cm³, more than 90% of total TRWP were separated from the sample matrix. TRWP concentrations in particulate matter collected in two road runoff treatment systems ranged from 0.38 to 150 mg TRWP/g. Differences in quantified TRWP contents of the two systems indicate changes in particle dynamics due to ageing and aggregation processes. The developed method allows TRWP determination in road runoff and in environments that are influenced by road traffic. The validated separation procedure can also be applied for TRWP characterization in future studies.

Development and testing of a fractionated filtration for sampling of microplastics in water.

A harmonization of sampling, sample preparation and detection is pivotal in order to obtain comparable data on microplastics (MP) in the environment. This paper develops and proposes a suitable sampling concept for waterbodies that considers different plastic specific properties and influencing factors in the environment. Both artificial water including defined MP fractions and the discharge of a wastewater treatment plant were used to verify the derived sampling procedure, sample preparation and the subsequent analysis of MP using thermal extraction-desorption gas chromatography - mass spectrometry (TED-GC-MS). A major finding of this paper is that an application of various particle size classes greatly improves the practical handling of the sampling equipment. Size classes also enable the TED-GC-MS to provide any data on the MP size distribution, a substantial sampling property affecting both the necessary sampling volume and the optimal sampling depth. In the artificial water with defined MP fractions, the recovery rates ranged from 80 to 110%, depending on the different MP types and MP size classes. In the treated wastewater, we found both polyethylene and polystyrene in different size classes and quantities.

High-throughput NIR spectroscopic (NIRS) detection of microplastics in soil.

The increasing pollution of terrestrial and aquatic ecosystems with plastic debris leads to the accumulation of microscopic plastic particles of still unknown amount. To monitor the degree of contamination, analytical methods are urgently needed, which help to quantify microplastics (MP). Currently, time-costly purified materials enriched on filters are investigated both by micro-infrared spectroscopy and/or micro-Raman. Although yielding precise results, these techniques are time consuming, and are restricted to the analysis of a small part of the sample in the order of few micrograms. To overcome these problems, we tested a macroscopic dimensioned near-infrared (NIR) process-spectroscopic method in combination with chemometrics. For calibration, artificial MP/ soil mixtures containing defined ratios of polyethylene, polyethylene terephthalate, polypropylene, and polystyrene with diameters < 125 µm were prepared and measured by a process FT-NIR spectrometer equipped with a fiber-optic reflection probe. The resulting spectra were processed by chemometric models including support vector machine regression (SVR), and partial least squares discriminant analysis (PLS-DA). Validation of models by MP mixtures, MP-free soils, and real-world samples, e.g., fermenter residue, suggests a reliable detection and a possible classification of MP at levels above 0.5 to 1.0 mass% depending on the polymer. The benefit of the combined NIRS chemometric approach lies in the rapid assessment whether soil contains MP, without any chemical pretreatment. The method can be used with larger sample volumes and even allows for an online prediction and thus meets the demand of a high-throughput method.