Laura J. Carter

Uptake and depuration of pharmaceuticals in aquatic invertebrates.

The uptake and depuration of a range of pharmaceuticals in the freshwater shrimp (Gammarus pulex) and the water boatman (Notonecta glauca) was studied. For one compound, studies were also done using the freshwater snail Planobarius corneus. In G. pulex, bioconcentration factors (BCFs) ranged from 4.6 to 185,900 and increased in the order moclobemide < 5-fluoruracil < carbamazepine < diazepam < carvedilol < fluoxetine. In N. glauca BCFs ranged from 0.1 to 1.6 and increased in the order 5-fluorouracil < carbamazepine < moclobemide < diazepam < fluoxetine < carvedilol. For P. corneus, the BCF for carvedilol was 57.3. The differences in degree of uptake across the three organisms may be due to differences in mode of respiration, behaviour and the pH of the test system. BCFs of the pharmaceuticals for each organism were correlated to the pH-corrected liposome-water partition coefficient of the pharmaceuticals.

Fate and Uptake of Pharmaceuticals in Soil–Plant Systems

Pharmaceuticals have been detected in the soil environment where there is the potential for uptake into crops. This study explored the fate and uptake of pharmaceuticals (carbamazepine, diclofenac, fluoxetine, propranolol, sulfamethazine) and a personal care product (triclosan) in soil–plant systems using radish (Raphanus sativus) and ryegrass (Lolium perenne). Five of the six chemicals were detected in plant tissue. Carbamazepine was taken up to the greatest extent in both the radish (52 μg/g) and ryegrass (33 μg/g), whereas sulfamethazine uptake was below the limit of quantitation (LOQ) (<0.01 μg/g). In the soil, concentrations of diclofenac and sulfamethazine dropped below the LOQ after 7 days. However, all pharmaceuticals were still detectable in the pore water at the end of the experiment. The results demonstrate the ability of plant species to accumulate pharmaceuticals from soils with uptake apparently specific to both plant species and chemical. Results can be partly explained by the hydrophobicity and extent of ionization of each chemical in the soil.

Fate and Uptake of Pharmaceuticals in Soil−Earthworm Systems

Pharmaceuticals present a potential threat to soil organisms, yet our understanding of their fate and uptake in soil systems is limited. This study therefore investigated the fate and uptake of 14C-labeled carbamazepine, diclofenac, fluoxetine, and orlistat in soil–earthworm systems. Sorption coefficients increased in the order of carbamazepine < diclofenac < fluoxetine < orlistat. Dissipation of 14C varied by compound, and for orlistat, there was evidence of formation of nonextractable residues. Uptake of 14C was seen for all compounds. Depuration studies showed complete elimination of 14C for carbamazepine and fluoxetine treatments and partial elimination for orlistat and diclofenac, with greater than 30% of the 14C remaining in the tissue at the end of the experiment. Pore-water-based bioconcentration factors (BCFs), based on uptake and elimination of 14C, increased in the order carbamazepine < diclofenac < fluoxetine and orlistat. Liquid chromatography–tandem mass spectrometry and liquid chromatography–Fourier transform mass spectrometry indicated that the observed uptake in the fluoxetine and carbamazepine treatments was due to the parent compounds but that diclofenac was degraded in the test system so uptake was due to unidentifiable transformation products. Comparison of our data with outputs of quantitative structure−activity relationships for estimating BCFs in worms showed that these models tend to overestimate pharmaceutical BCFs so new models are needed.

Uptake of Pharmaceuticals Influences Plant Development and Affects Nutrient and Hormone Homeostases.

The detection of a range of active pharmaceutical ingredients (APIs) in the soil environment has led to a number of publications demonstrating uptake by crops, however very few studies have explored the potential for impacts on plant development as a result of API uptake. This study investigated the effect of carbamazepine and verapamil (0.005-10 mg/kg) on a range of plant responses in zucchini (Cucurbita pepo). Uptake increased in a dose-dependent manner, with maximum leaf concentrations of 821.9 and 2.2 mg/kg for carbamazepine and verapamil, respectively. Increased carbamazepine uptake by zucchini resulted in a decrease in above (<60%) and below (<30%) ground biomass compared to the controls (p < 0.05). At soil concentrations >4 mg/kg the mature leaves suffered from burnt edges and white spots as well as a reduction in photosynthetic pigments but no such effects were seen for verapamil. For both APIs, further investigations revealed significant differences in the concentrations of selected plant hormones (auxins, cytokinins, abscisic acid and jasmonates), and in the nutrient composition of the leaves in comparison to the controls (p < 0.05). This is some of the first research to demonstrate that the exposure of plants to APIs is likely to cause impacts on plant development with unknown implications.

Minimised bioconcentration tests: a useful tool for assessing chemical uptake into terrestrial and aquatic invertebrates?

Current guidelines for determining bioconcentration factors (BCF) and uptake and depuration rate constants require labor intensive studies with large numbers of organisms. A minimized approach has recently been proposed for fish BCF studies but its applicability to other taxonomic groups is unknown. In this study, we therefore evaluate the use of the minimized approach for estimating BCF and uptake and depuration rate constants for chemicals in aquatic and terrestrial invertebrates. Data from a range of previous BCF studies were resampled to calculate BCFs and rate constants using the minimized method. The resulting values were then compared to values obtained using full study designs. Results demonstrated a good correlation for uptake rate constants, a poor correlation for depuration rate constants and a very good correlation between the BCFs obtained using the traditional and minimized approach for a variety of organic compounds. The minimized approach therefore has merit in deriving bioconcentration factors and uptake rate constants but may not be appropriate for deriving depuration rate constants for use in, for example, toxico-kinetic toxico-dynamic modeling. The approach uses up to 70% fewer organisms, requires less labor and has lower analytical costs. The minimized design therefore could be a valuable approach for running large multifactorial studies to assess bioconcentration of the plethora of chemicals that occur in the environment into the many taxonomic groups that occur in the environment. The approach should therefore help in accelerating the development of our understanding of factors and processes affecting uptake of chemicals into organisms in the environment.

Effects of soil properties on the uptake of pharmaceuticals into earthworms

Pharmaceuticals can enter the soil environment when animal slurries and sewage sludge are applied to land as a fertiliser or during irrigation with contaminated water. These pharmaceuticals may then be taken up by soil organisms possibly resulting in toxic effects and/or exposure of organisms higher up the food chain. This study investigated the influence of soil properties on the uptake and depuration of pharmaceuticals (carbamazepine, diclofenac, fluoxetine and orlistat) in the earthworm Eisenia fetida. The uptake and accumulation of pharmaceuticals into E. fetida changed depending on soil type. Orlistat exhibited the highest pore water based bioconcentration factors (BCFs) and displayed the largest differences between soil types with BCFs ranging between 30.5 and 115.9. For carbamazepine, diclofenac and fluoxetine BCFs ranged between 1.1 and 1.6, 7.0 and 69.6 and 14.1 and 20.4 respectively. Additional analysis demonstrated that in certain treatments the presence of these chemicals in the soil matrices changed the soil pH over time, with a statistically significant pH difference to control samples. The internal pH of E. fetida also changed as a result of incubation in pharmaceutically spiked soil, in comparison to the control earthworms. These results demonstrate that a combination of soil properties and pharmaceutical physico-chemical properties are important in terms of predicting pharmaceutical uptake in terrestrial systems and that pharmaceuticals can modify soil and internal earthworm chemistry which may hold wider implications for risk assessment.

Temporal and spatial variation in pharmaceutical concentrations in an urban river system

Many studies have quantified pharmaceuticals in the environment, few however, have incorporated detailed temporal and spatial variability due to associated costs in terms of time and materials. Here, we target 33 physico-chemically diverse pharmaceuticals in a spatiotemporal exposure study into the occurrence of pharmaceuticals in the wastewater system and the Rivers Ouse and Foss (two diverse river systems) in the city of York, UK. Removal rates in two of the WWTPs sampled (a conventional activated sludge (CAS) and trickling filter plant) ranged from not eliminated (carbamazepine) to >99% (paracetamol). Data comparisons indicate that pharmaceutical exposures in river systems are highly variable regionally, in part due to variability in prescribing practices, hydrology, wastewater management, and urbanisation and that select annual median pharmaceutical concentrations observed in this study were higher than those previously observed in the European Union and Asia thus far. Significant spatial variability was found between all sites in both river systems, while seasonal variability was significant for 86% and 50% of compounds in the River Foss and Ouse, respectively. Seasonal variations in flow, in-stream attenuation, usage and septic effluent releases are suspected drivers behind some of the observed temporal exposure variability. When the data were used to evaluate a simple environmental exposure model for pharmaceuticals, mean ratios of predicted environmental concentrations (PECs), obtained using the model, to measured environmental concentrations (MECs) were 0.51 and 0.04 for the River Foss and River Ouse, respectively. Such PEC/MEC ratios indicate that the model underestimates actual concentrations in both river systems, but to a much greater extent in the larger River Ouse.

Sorption, plant uptake and metabolism of benzodiazepines

Reuse of treated wastewater for irrigation of crops is growing in arid and semi-arid regions, whilst increasing amounts of biosolids are being applied to fields to improve agricultural outputs. Due to incomplete removal in the wastewater treatment processes, pharmaceuticals present in treated wastewater and biosolids can contaminate soil systems. Benzodiazepines are a widely used class of pharmaceuticals that are released following wastewater treatment. Benzodiazepines are represented by a class of compounds with a range of physicochemical properties and this study was therefore designed to evaluate the influence of soil properties on the sorption behaviour and subsequent uptake of seven benzodiazepines (chlordiazepoxide, clonazepam, diazepam, flurazepam, oxazepam, temazepam and triazolam) in two plant species. The sorption and desorption behaviour of benzodiazepines was strongly influenced by soil type and hydrophobicity of the chemical. The partitioning behaviour of these chemicals in soil was a key controller of the uptake and accumulation of benzodiazepines by radish (Raphanus sativus) and silverbeet (Beta vulgaris). Benzodiazepines such as oxazepam that were neutral, had low sorption coefficients (Kd) or had pH-adjusted log octanol-water partition coefficients (log Dow, pH6.3) values close to 2 had the greatest extent of uptake. Conversely, benzodiazepines such as flurazepam that had an ionised functional groups and greater Kd values had comparatively limited accumulation in the selected plant species. Results also revealed active in-plant metabolism of benzodiazepines, potentially analogous to the known metabolic transformation pathway of benzodiazepines in humans. Along with this observed biological transformation of benzodiazepines in exposed plants, previously work has established the widespread presence of the plant signalling molecule γ-amino butyric acid (GABA), which is specifically modulated by benzodiazepines in humans. This highlights the need for further assessment of the potential for biological activity of benzodiazepines following their plant uptake.

Application of prioritization approaches to optimize environmental monitoring and testing of pharmaceuticals

ABSTRACT Pharmaceuticals are ubiquitous in the natural environment with concentrations expected to rise as human population increases. Environmental risk assessments are available for a small portion of pharmaceuticals in use, raising concerns over the potential risks posed by other drugs that have little or no data. With >1900 active pharmaceutical ingredients in use, it would be a major task to test all of the compounds with little or no data. Desk-based prioritization studies provide a potential solution by identifying those substances that are likely to pose the greatest risk to the environment and which, therefore, need to be considered a priority for further study. The aim of this review was to (1) provide an overview of different prioritization exercises performed for pharmaceuticals in the environment and the results obtained; and (2) propose a new holistic risk-based prioritization framework for drugs in the environment. The suggested models to underpin this framework are discussed in terms of validity and applicability. The availability of data required to run the models was assessed and data gaps identified. The implementation of this framework may harmonize pharmaceutical prioritization efforts and ensure that, in the future, experimental resources are focused on molecules, endpoints, and environmental compartments that are biologically relevant.

Bioaccumulation, uptake, and toxicity of carbamazepine in soil–plant systems

Since the detection of active pharmaceutical ingredients (APIs) in various environmental media, research has explored the potential uptake and toxicity of these chemicals to species inhabiting these matrices. Specifically, pharmaceuticals, including the antiepileptic API carbamazepine (CBZ), are taken up from soil by a range of plants. Many short-term studies have also suggested that certain APIs induce toxicity in plants. However, the effects of APIs on fruiting plants remain relatively unexplored. The present study investigated the uptake, bioaccumulation, and toxicity of CBZ in Cucurbita pepo (zucchini) from seed to full maturity across a range of CBZ exposure concentrations in soil (0.1-20 mg/kg). Results of biomass, chlorophyll, starch and total nitrogen (N) concentration in C. pepo indicated toxicity at soil concentrations of ≥10 mg/kg. There were clear visual indications of increasing toxicity on leaves, including chlorosis and necrosis, from soil concentrations of 1 up to 20 mg/kg. The present study also revealed novel insights into the effect of CBZ accumulation on C. pepo fruiting: female C. pepo flowers were unable to set fruit when leaf concentrations were ≥14 mg/kg. These findings may have implications for future agricultural productivity in areas where reclaimed wastewater containing APIs is a source of irrigation. Detectable CBZ concentrations were found in edible C. pepo fruit, indicating the possibility of trophic transfer. Environ Toxicol Chem 2018;37:1122-1130.