Stable isotopes (15N) facilitate non-invasive labelling of large quantities of macroinvertebrates across different species and feeding types.

While macroinvertebrate dispersal operates at the individual level, predictions of their dispersal capabilities often rely on indirect proxies rather than direct measurements. To gain insight into the dispersal of individual specimens, it is crucial to mark (label) and capture individuals. Isotopic enrichment with 15N is a non-invasive method with the potential of labelling large quantities of macroinvertebrates. While the analysis of 15N is widely utilised in food web studies, knowledge on the specific utility of isotopic enrichment with 15N for mass labelling of macroinvertebrate individuals across different taxa and feeding types is limited. Previous studies have focused on single species and feeding types, leaving gaps in our understanding of the broader applicability of this method. Therefore, this study aimed to test and compare isotopic mass enrichment across several macroinvertebrate taxa and feeding types. We released 15NH4Cl at five stream reaches in North-Rhine Westphalia, Germany, and successfully enriched 12 distinct macroinvertebrate taxa (Crustacea and Insecta). Significant enrichment was achieved in active and passive filter feeders, grazers, shredders and predators, and predominantly showed positive correlations with the enrichment of the taxa's main food sources phytobenthos and particulate organic matter. Enrichment levels rose rapidly and peaked at distances between 50 m and 300 m downstream of the isotopic inlet; significant enrichment occurred up to 2000 m downstream of the isotopic inlet in all feeding types. Macroinvertebrate density estimates on the stream bottom averaged to a total of approximately 3.4 million labelled individuals of the 12 investigated taxa, thus showing the high potential of isotopic (15N) enrichment as a non-invasive method applicable for mass labelling across different macroinvertebrate feeding types. Hence, isotopic enrichment can greatly assist the analysis of macroinvertebrate dispersal through mark-and-recapture experiments, as it allows to measure the movement at the level of individual specimens.

Evaluation of GC-EI&CI-TOFMS for Nontarget Analysis of Industrial Wastewater Using Hydrophilic-Lipophilic-Balanced SPME.

The evaluation of nontarget analysis (NTA) techniques for the monitoring of wastewater is important as wastewater is an anthropogenic pollution source for aquatic ecosystems and a threat to human and environmental health. This study presents the proof-of-concept NTA of industrial wastewater samples. A prototype hydrophilic-lipophilic-balanced (HLB) SPME and gas chromatography interfaced with time-of-flight high-resolution mass spectrometry (GC-TOFMS) with electron ionization (EI) and chemical ionization (CI) in parallel are employed. The HLB-SPME consists of a poly(divinylbenzene-co-N-vinylpyrrolidone) structure, allowing the extraction of hydrophilic as well as lipophilic substances. As the combination of parallel CI and EI data provides a comprehensive data set as a unique feature, this study is strongly focused on the compound identification procedure and confidence reporting of exemplary substances. Furthermore, the use of three different CI reagent ions, including [N2H]+/[N4H]+, [H3O]+, and [NH4]+, enables a broad range of analytes to be ionized in terms of selectivity and softness. The complementary information provided by EI and CI data allows a level 3 identification or higher in 69% of cases. The polarity coverage based on the physicochemical properties of the analytes (such as volatility, water solubility, hydrophilicity, and lipophilicity) was visualized by using Henry's law and octanol-water partitioning constants. In conclusion, the presented approach is shown to be valuable for water analysis and allows enhanced and accelerated compound identification compared to utilizing only one type of ionization.

Dispersive liquid-liquid microextraction as a novel enrichment approach for compound-specific carbon isotope analysis of chlorinated phenols.

Compound-specific isotope analysis (CSIA) via gas chromatography-isotope ratio mass spectrometry (GC-IRMS) is a potent tool to elucidate the fate of (semi-)volatile organic contaminants in technical and environmental systems. Yet, due to the comparatively low sensitivity of IRMS, an enrichment step prior to analysis often is inevitable. A promising approach for fast as well as economic analyte extraction and preconcentration prior to CSIA is dispersive liquid-liquid microextraction (DLLME) - a well-established technique in concentration analysis of contaminants from aqueous samples. Here, we present and evaluate the first DLLME method for GC-IRMS exemplified by the analysis of chlorinated phenols (4-chlorophenol, 2,4-dichlorophenol, and 2,4,6-trichlorophenol) as model compounds. The analytes were simultaneously acetylated with acetic anhydride and extracted from the aqueous phase using a binary solvent mixture of acetone and tetrachloroethylene. With this method, reproducible δ13C values were achieved with errors ≤ 0.6‰ (n = 3) for aqueous concentrations down to 100 µg L-1. With preconcentration factors between 130 and 220, the method outperformed conventional liquid-liquid extraction in terms of sample preparation time and resource consumption with comparable reproducibility. Furthermore, we have demonstrated the suitability of the method (i) for the extraction of the analytes from a spiked river water sample and (ii) to quantify kinetic carbon isotope effect for 2,4,6-trichlorophenol during reduction with zero-valent zinc in a laboratory batch experiment. The presented work shows for the first time the potential of DLLME for analyte enrichment prior to CSIA and paves the way for further developments, such as the extraction of other compounds or scaling up to larger sample volumes.

Isotope-labeling in situ derivatization and HS-SPME arrow GC-MS/MS for simultaneous determination of fatty acids and fatty acid methyl esters in aqueous matrices.

Fatty acids (FAs) and fatty acid methyl esters (FAMEs) co-occur in many samples, and analysis of both substance classes is frequently of high interest. To this end, this study introduces the first method for simultaneous determination of FAs and FAMEs including fully automated solvent-free solid-phase microextraction (SPME) arrow headspace extraction combined with isotope-labeling in situ FA derivatization with deuterated methanol (CD3OD). By using the chromatographic isotope effect (ΔRt = 0.03 min) and the + 3 m/z mass shift, FAs can be selectively differentiated from the FAMEs during gas chromatography tandem-mass spectrometry (GC-MS/MS) operated in the multiple reaction monitoring (MRM) aquisition mode. Additionally, an approach is presented to predict the retention times of deuterated compounds. Optimization of the derivatization conditions was accomplished by design of experiments and found to be 20 min, 50 °C, 4 v/v% CD3OD, and pH 2.1. During method validation, FAs and FAMEs were calibrated in different concentration ranges by standard addition in five real matrices and ultrapure water leading to good linearities and method detection limits for FAs ranging from 1-30 µg L-1 and for FAMEs from 0.003-0.72 µg L-1. FAs and FAMEs were detected in real samples from surface water, wastewater treatment plant effluent, and three different bioreactor samples and could be quantified in concentrations ranging from 2-1056 µg L-1 for FAs and 0.01-14 µg L-1 for FAMEs.

Improving greenness and sustainability of standard analytical methods by microextraction techniques: A critical review.

Since environmental awareness has increased in analytical chemistry, the demand for green sample preparation methods continues to grow. Microextractions such as solid-phase microextraction (SPME) and liquid-phase microextraction (LPME) miniaturize the pre-concentration step and are a more sustainable alternative to conventional large-scale extractions. However, the integration of microextractions in standard and routine analysis methods is rare, although these applications are used most frequently and have a role model function. Therefore, it is important to highlight that microextractions are capable to replace large-scale extractions in standard and routine methods. This review discusses the greenness, benefits, and drawbacks of the most common LPME and SPME variants compatible with gas chromatography based on the following key evaluation principles: Automation, solvent consumption, hazards, reusability, energy consumption, time efficiency, and handling. Furthermore, the need to integrate microextractions into standard and routine analytical methods is presented by using method greenness evaluation metrics AGREE, AGREEprep, and GAPI applied to USEPA methods and their replacements.

The Asymmetric Response Concept explains ecological consequences of multiple stressor exposure and release.

Our capacity to predict trajectories of ecosystem degradation and recovery is limited, especially when impairments are caused by multiple stressors. Recovery may be fast or slow and either complete or partial, sometimes result in novel ecosystem states or even fail completely. Here, we introduce the Asymmetric Response Concept (ARC) that provides a basis for exploring and predicting the pace and magnitude of ecological responses to, and release from, multiple stressors. The ARC holds that three key mechanisms govern population, community and ecosystem trajectories. Stress tolerance is the main mechanism determining responses to increasing stressor intensity, whereas dispersal and biotic interactions predominantly govern responses to the release from stressors. The shifting importance of these mechanisms creates asymmetries between the ecological trajectories that follow increasing and decreasing stressor intensities. This recognition helps to understand multiple stressor impacts and to predict which measures will restore communities that are resistant to restoration.

A new technique to study nutrient flow in host-parasite systems by carbon stable isotope analysis of amino acids and glucose.

Stable isotope analysis of individual compounds is emerging as a powerful tool to study nutrient origin and conversion in host-parasite systems. We measured the carbon isotope composition of amino acids and glucose in the cestode Schistocephalus solidus and in liver and muscle tissues of its second intermediate host, the three-spined stickleback (Gasterosteus aculeatus), over the course of 90 days in a controlled infection experiment. Similar linear regressions of δ13C values over time and low trophic fractionation of essential amino acids indicate that the parasite assimilates nutrients from sources closely connected to the liver metabolism of its host. Biosynthesis of glucose in the parasite might occur from the glucogenic precursors alanine, asparagine and glutamine and with an isotope fractionation of - 2 to - 3 ‰ from enzymatic reactions, while trophic fractionation of glycine, serine and threonine could be interpreted as extensive nutrient conversion to fuel parasitic growth through one-carbon metabolism. Trophic fractionation of amino acids between sticklebacks and their diets was slightly increased in infected compared to uninfected individuals, which could be caused by increased (immune-) metabolic activities due to parasitic infection. Our results show that compound-specific stable isotope analysis has unique opportunities to study host and parasite physiology.

Insights into amino acid fractionation and incorporation by compound-specific carbon isotope analysis of three-spined sticklebacks.

Interpretation of stable isotope data is of upmost importance in ecology to build sound models for the study of animal diets, migration patterns and physiology. However, our understanding of stable isotope fractionation and incorporation into consumer tissues is still limited. We therefore measured the δ13C values of individual amino acids over time from muscle and liver tissue of three-spined sticklebacks (Gasterosteus aculeatus) on a high protein diet. The δ13C values of amino acids in the liver quickly responded to small shifts of under ± 2.0‰ in dietary stable isotope compositions on 30-day intervals. We found on average no trophic fractionation in pooled essential (muscle, liver) and non-essential (muscle) amino acids. Negative Δδ13C values of - 0.7 ± 1.3‰ were observed for pooled non-essential (liver) amino acids and might indicate biosynthesis from small amounts of dietary lipids. Trophic fractionation of individual amino acids is reported and discussed, including unusual Δδ13C values of over + 4.9 ± 1.4‰ for histidine. Arginine and lysine showed the lowest trophic fractionation on individual sampling days and might be useful proxies for dietary sources on short time scales. We suggest further investigations using isotopically enriched materials to facilitate the correct interpretation of ecological field data.

Optimization and automation of rapid and selective analysis of fatty acid methyl esters from aqueous samples by headspace SPME arrow extraction followed by GC-MS/MS analysis.

The analysis of fatty acid methyl esters (FAMEs) is of high relevance for monitoring and control of various industrial processes and biological systems. In this study, a novel, green analytical approach for the determination of 24 FAMEs from aqueous samples is proposed, which is based on a headspace solid-phase microextraction (SPME) arrow followed by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). The method was substantially accelerated to a run time of 44 min per sample by thorough optimization and automation of the relevant parameters. The limiting parameters, mostly based on expediting equilibrium attainment, were found to be parameters of extraction: material, pH, time, and temperature, which were optimized to divinylbenzene polydimethylsiloxane (DVB-PDMS), pH 2, 20 min, and 70 °C, respectively. The optimization and automation of the method led to low method detection limits (9-437 ng L-1) and high selectivity. Evaluation of the method on real samples was done by analyzing the aqueous phase of a bioreactor, whereby the matrix effect could be greatly reduced due to dilution and headspace sampling. The rapid, sensitive, selective, and matrix-reduced approach is found to be not only a novel method for water analysis but is promising for further applications, e.g., with solid and gaseous samples containing FAMEs.

In-tube dynamic extraction for analysis of volatile organic compounds in honey samples.

Honey is the oldest and nowadays widely used natural sweetener for food worldwide. Its composition is associated with its botanical and geographical origin and honey is often mislabeled and has a high potential for food fraud. Thus, quick easy and sensitive analyses are required. For the first time, we developed and applied an automated, fast, sensitive and robust, in-tube extraction dynamic headspace in-tube extraction-dynamic headspace (ITEX-DHS) method for a variety of Honey containing VOCs in connection with GC-MS. Another advantage of ITEX is, that it is a green analytical solventless method. The method provides very low method detection limits (MDL) from 0.8 to 47 ng g-1 for VOCs in honey samples as well as very good repeatabilities with averages below 9 % RSD. Recoveries are between 83 and 100 %. Only octanal possess a repeatability 13 % and a recovery of 62 % due to its high polarity. 38 honey samples were measured after method validation. Four acacia honeys (A), six forest honeys (F) and 22 blossom honeys (B). The type of six honeys was not known (U) but could be predicted with the help of a linear discriminant analysis (LDA). The LDA was carried out with the three groups (A, B, F) leading to a proportion of correct predictions of 90.6 %. With the help of a scatterplot, two of the unknown samples were classified as forest honeys and four of them as blossom honeys.

Aromatic amines contents of cigarette butts: Fresh and aged cigarette butts vs unsmoked cigarette.

Cigarette butts (CBs) are some of the most abundant waste items in the environment and may contain high levels of different toxic chemicals, such as aromatic amines (AAs). However, to this date, there is no comprehensive study on the role of CBs in the emission of AAs into the environment. The present study investigated for the first time the concentration levels of 10 primary aromatic amines (PAAs), including ANL, p-TOL, m-TOL, o-TOL, 2,6-DMA, o-ASD, 2-NA, 1-NA, 3-ABP, and 4-ABP that were measured and compared in unsmoked cigarette, freshly smoked CBs, and CBs collected from urban streets (named here aged CBs). The mean levels of ∑PAAs in different sample categories were statistically significantly different and the mean level order was as freshly smoked CBs > aged CBs > unsmoked cigarette with the values of 3.43, 2.12 µg g-1, and 0.28 µg g-1, respectively. The levels of ∑PAAs, ANL, o-ASD, 2,6-DMA, 2-NA, and ∑TOL dramatically increased by 12.26, 4.05, 8.46, 10.41, 4.78, and 28.84 times, respectively, right after smoking comparing the freshly smoked CBs samples and unsmoked cigarette. The results also showed a substantial decrease in the levels of PAAs (except o-ASD) in the aged CBs samples compared to freshly smoked CBs. The levels of ∑PAAs, o-ASD, 2,6-DMA, ∑TOL, ANL, 2-NA, 1-NA, and ∑ABP decreased 1.62, 1.09, 1.91, 3.20, 3.42, 2.63, 2.00, and 1.88 times, respectively. Considering the average PAAS content and estimated CBs littered worldwide every year, freshly smoked CBs can theoretically emit 2.9 tons of ∑PAAs into the environment annually. Considering other chemicals that may also be released into the environment via CBs (beside PAAs), we can consider CBs as a critical source of toxic compounds into the environment and water bodies.

Two Pathways Compete in the Mn(II)-Catalyzed Oxidation of Aminotrismethylene Phosphonate (ATMP).

Mn(II)-catalyzed oxidation by molecular oxygen is considered a relevant process for the environmental fate of aminopolyphosphonate chelating agents such as aminotrismethylene phosphonate (ATMP). However, the potential roles of Mn(III)ATMP-species in the underlying transformation mechanisms are not fully understood. We combined kinetic studies, compound-specific stable carbon isotope analysis, and equilibrium speciation modeling to shed light on the significance of such Mn-ATMP species for the overall ATMP oxidation by molecular oxygen. The fraction of ATMP complexed with Mn(II) inversely correlated with both (i) the Mn(II)-normalized transformation rate constants of ATMP and (ii) the observed carbon isotope enrichment factors (εc-values). These findings provide evidence for two parallel ATMP transformation pathways exhibiting distinctly different reaction kinetics and carbon isotope fractionation: (i) oxidation of ATMP present in Mn(III)ATMP complexes (εc ≈ -10 ‰) and (ii) oxidation of free ATMP by such Mn(III)ATMP species (εc ≈ -1 ‰) in a catalytic cycle. The higher reaction rate of the latter pathway implies that aminopolyphosphonates can be trapped in catalytic Mn-complexes before being transformed and suggests that Mn(III)ATMP might be a potent oxidant also for other reducible solutes in aqueous environments.

How to Couple LC-IRMS with HRMS─A Proof-of-Concept Study.

Compound-specific stable isotope analysis (CSIA) is a unique analytical technique for determining small variations in isotope ratios of light isotopes in analytes from complex mixtures. A problem of CSIA using gas chromatography (GC) and liquid chromatography-isotope ratio mass spectrometry (LC-IRMS) is that any structural information of the analytes is lost due to the processes involved in determining the isotope ratio. To obtain the isotopic composition of, for example, carbon from organic compounds, all carbon in each analyte is quantitatively converted to CO2. For GC-IRMS, open split GC-IRMS-MS couplings have been described that allow additional acquisition of structural information of analytes and interferences. Structural analysis using LC-IRMS is more difficult and requires additional technical and instrumental efforts. In this study, LC was combined for the first time with simultaneous analysis by IRMS and high-resolution mass spectrometry (HRMS), enabling the direct identification of unknown or coeluting species. We have thoroughly investigated and optimized the coupling and showed how technical problems, arising from instrumental conditions, can be overcome. To this end, it was successfully demonstrated that a consistent split ratio between IRMS and HRMS could be obtained using a variable postcolumn flow splitter. This coupling provided reproducible results in terms of resulting peak areas, isotope values, and retention time differences for the two mass spectrometer systems. To demonstrate the applicability of the coupling, we chose to address an important question regarding the purity of international isotope standards. In this context, we were able to confirm that the USGS41 reference material indeed contains substantial amounts of pyroglutamic acid as suggested previously in the literature. Moreover, the replacement material, USGS41a, still has significant amounts of pyroglutamic acid as impurity, rendering some caution necessary when using this material for isotopic calibration.

Linking reaction rate constants and isotope fractionation of ozonation reactions using phenols as probes.

Ozonation is nowadays a widely used method in drinking water treatment for disinfection and pollutant control. However, transformation products of ozonation can be more toxic than their parent compounds. Therefore, the knowledge of the reaction mechanisms and product formation is essential for a safe application. Different analytical methods such as high-resolution mass spectrometry (HRMS) and compound-specific isotope analysis (CSIA) can be applied to elucidate products and primary attack positions of oxidation agents such as ozone. During the investigation of the ozonation of phenolic compounds in water by CSIA, a reaction rate depending carbon isotope fractionation was observed. The fractionation strongly depends on the phenol speciation. With decreasing pH values and reaction rates <105 M-1 s-1, the isotope enrichment factor ε increases (ε is between -5.2 and -1.0‰). For faster reactions (>105 M-1 s-1), the carbon isotope enrichment was not significant anymore (ε is between -1.0 and 0‰). Based on these data a concept to correlate isotope enrichment factors with kinetic data for aromatic compounds is proposed. The additional investigation of aliphatic double and triple bonds did not fit this correlation suggesting different rate-limiting steps. However, double and triple bond showed a similar enrichment factor, which implies the same rate-limiting step in the reaction with ozone, the monodentate addition of ozone.

Eye fluke infection changes diet composition in juvenile European perch (Perca fluviatilis).

Intraspecific diet specialization, usually driven by resource availability, competition and predation, is common in natural populations. However, the role of parasites on diet specialization of their hosts has rarely been studied. Eye flukes can impair vision ability of their hosts and have been associated with alterations of fish feeding behavior. Here it was assessed whether European perch (Perca fluviatilis) alter their diet composition as a consequence of infection with eye flukes. Young-of-the-year (YOY) perch from temperate Lake Müggelsee (Berlin, Germany) were sampled in two years, eye flukes counted and fish diet was evaluated using both stomach content and stable isotope analyses. Perch diet was dominated by zooplankton and benthic macroinvertebrates. Both methods indicated that with increasing eye fluke infection intensity fish had a more selective diet, feeding mainly on the benthic macroinvertebrate Dikerogammarus villosus, while less intensively infected fish appeared to be generalist feeders showing no preference for any particular prey type. Our results show that infection with eye flukes can indirectly affect interaction of the host with lower trophic levels by altering the diet composition and highlight the underestimated role of parasites in food web studies.

BTEX compounds leachates from cigarette butts into water environment: A primary study.

Cigarette butts (CBs) are the most abundant types of litter in the environment and may contain toxic chemicals such as BTEX that pose serious risks to the water bodies and health of aquatic organisms. So far there is no systematic study on BTEX compounds (benzene, toluene, ethylbenzene, o-xylene, and p-xylene) leaching from CBs into water environments. In this work, the leaching concentrations of BTEX compounds in deionized water (DW) and river water (RW) samples were studied for the first time. The mean concentrations of benzene, toluene, ethylbenzene, p-xylene, and o-xylene at contact times of 15 min to 1 day in water samples ranged from 0.13 to 0.18, 0.39-0.9, 0.11-0.25, 0.12-0.38, and 0.09-0.19 µgL-1 respectively. Benzene, toluene, ethylbenzene, o-xylene and p-xylene were detected at all contact times in both DW and RW samples. There were no significant differences of the leachate levels of BTEX compounds between DW and RW samples. The highest and lowest mean concentration levels in both DW and RW samples were determined for toluene and o-xylene respectively. The time after smoking had a significant effect on BTEX levels in leachates. The concentration levels of benzene, toluene, ethylbenzene, o-xylene and p-xylene leachates in water samples, after only 15 min, were reduced by 100, 93, 70, 68, and 59 percent respectively. Our data revealed that leached concentrations of benzene did not exceed the Water Framework Directive (WFD) guidelines, but with regard to the amount of CBs littered each year and other toxic chemicals contents of CBs this can still be a threat for aquatic creatures and possibly humans as well. Further studies are needed to cover the knowledge gap on the toxic leachates from CBs into water systems.

Optimization and validation of automated solid-phase microextraction arrow technique for determination of phosphorus flame retardants in water.

In the present work, a very sensitive and fully automated direct immersion PAL SPME Arrow procedure, coupled with GC-MS, has been developed and validated for determination of nine phosphorus flame retardants in different types of water samples (river, drinking and rainwater). PDMS/DVB was selected among three commercially available SPME Arrows (PDMS/DVB, DVB/PDMS/CWR and PDMS/CWR), since it resulted in the best sensitivity. The important experimental parameters were optimized via a central composite design response surface methodology and as result, extraction time of 65 min, extraction temperature of 80 °C and added salt concentration of 19% (w/v), were selected as the optimum values. The optimized method showed linear response over the calibration range (2 - 500 ng L-1), with R2-values higher than 0.9937. The precision (RSD%) measured by replicate analyses (n = 7) was estimated at 2 and 100 ng L-1 and was less than 29% and 21%, respectively. The LOQ of PAL SPME Arrow, calculated as S/N = 10, was between 0.2 and 1.2 ng L-1 (for triphenyl phosphate and tris-(1­chloro­2-propyl) phosphate, respectively) with extraction efficiencies between 5.9 and 31% (for tris-(1,3-dichloro-2-propyl) phosphate and tri-n­butyl phosphate, respectively). To assess the performance of the developed technique for real samples, two river water samples, tap water from two regions and a rainwater sample were analyzed. Most of the target analytes were observed in the river samples with concentrations of 1.0 - 250 ng L-1 and the obtained recoveries at 50 ng L-1 ranged between 60 and 107%. Considering the figures of merit of the optimized method, PAL SPME Arrow-GC-MS showed to be the most sensitive analytical approach for determination of phosphorus flame retardants in water, with satisfying precision and accuracy, compared with conventional SPME-NPD, LLE-GC-MS and SPE-LC-MS/MS.

Stable isotope analysis spills the beans about spatial variance in trophic structure in a fish host - parasite system from the Vaal River System, South Africa.

Stable isotope analysis offers a unique tool for comparing trophic interactions and food web architecture in ecosystems based on analysis of stable isotope ratios of carbon (13C/12C) and nitrogen (15N/14N) in organisms. Clarias gariepinus were collected from six sites along the Vaal River, South Africa and were assessed for ectoparasites and endoparasites. Lamproglena clariae (Copepoda), Tetracampos ciliotheca and Proteocephalus glanduligerus (Cestoda), and larval Contracaecum sp. (Nematoda) were collected from the gills, intestine and mesenteries, respectively. Signatures of δ13C and δ15N were analysed in host muscle tissue and parasites using bulk stable isotope analysis. Variable stable isotope enrichment between parasites and host were observed; L. clariae and the host shared similar δ15N signatures and endoparasites being depleted in δ13C and δ15N relative to the host. Differences in stable isotope enrichment between parasites could be related to the feeding strategy of each parasite species collected. Geographic and spatial differences in enrichment of stable isotopes observed in hosts were mirrored by parasites. As parasites rely on a single host for meeting their nutritional demands, stable isotope variability in parasites relates to the dietary differences of host organisms and therefore variations in baseline stable isotope signatures of food items consumed by hosts.

Carbon Isotope Fractionation of Substituted Benzene Analogs during Oxidation with Ozone and Hydroxyl Radicals: How Should Experimental Data Be Interpreted?

Oxidative processes frequently contribute to organic pollutant degradation in natural and engineered systems, such as during the remediation of contaminated sites and in water treatment processes. Because a systematic characterization of abiotic reactions of organic pollutants with oxidants such as ozone or hydroxyl radicals by compound-specific stable isotope analysis (CSIA) is lacking, stable isotope-based approaches have rarely been applied for the elucidation of mechanisms of such transformations. Here, we investigated the carbon isotope fractionation associated with the oxidation of benzene and several methylated and methoxylated analogs, namely, toluene, three xylene isomers, mesitylene, and anisole, and determined their carbon isotope enrichments factors (εC) for reactions with ozone (εC = -3.6 to -4.6 ‰) and hydroxyl radicals (εC = 0.0 to -1.2‰). The differences in isotope fractionation can be used to elucidate the contribution of the reactions with ozone or hydroxyl radicals to overall transformation. Derivation of apparent kinetic isotope effects (AKIEs) for the reaction with ozone, however, was nontrivial due to challenges in assigning reactive positions in the probe compounds for the monodentate attack leading to an ozone adduct. We present several options for this step and compare the outcome to quantum chemical characterizations of ozone adducts. Our data show that a general assignment of reactive positions for reactions of ozone with aromatic carbons in ortho-, meta-, or para-positions is not feasible and that AKIEs of this reaction should be derived on a compound-by-compound basis.

Polycyclic aromatic hydrocarbons (PAHs) leachates from cigarette butts into water.

Cigarette butts (CBs) are the most common littered items in the environment and may contain high amounts of polycyclic aromatic hydrocarbons (PAHs) from incomplete tobacco leave burning. The potential relevance of PAHs stemming from CBs for aquatic systems remain unclear since to date there is no systematic study on PAHs leaching from CBs. Therefore, in this study the leaching concentrations of 16 EPA-PAHs (except benzo(ghi)perylene) in 3 different types of water were measured. The concentrations of ΣPAHs leachates from 4 h to 21 days ranged from 3.9 to 5.7, 3.3-5.5, and 3.0-5.0 µg L-1 for deionized, tap, and river waters, respectively. For all contact times, there were no substantial differences of the leachate concentrations of PAHs among different water types. Lighter PAHs had the highest concentrations among the detected PAHs and they were detected in the leachates already after 4 h. Concentrations of indeno(1,2,3-cd)pyrene, and dibenz(a,h)anthracene were below the limit of detection in all water samples at different contact times. At all contact times naphthalene and fluorene had the highest concentrations among the studied PAHs. Tap and river water samples with addition of sodium azide as chemical preservative contained significantly higher concentration of ΣPAHs. Our leaching data showed that leached concentrations of PAHs exceeded the Water Framework Directive (WFD) standards and considering the number of CBs annually littered this may pose a risk to aquatic organisms and potentially also humans.