LC-MS/MS analysis of pharmaceuticals in the Irish aquatic environment and the potential for human exposure

Reports concerning the quantitative analysis of pharmaceuticals in marine ecosystems are somewhat limited. It is necessary to determine pharmaceutical fate and assess any potential risk of exposure to aquatic species and ultimately, seafood consumers. However, in Ireland very little research has been carried out to determine the presence of pharmaceutical residues in the aquatic environment. The research carried out investigates the occurrence of pharmaceuticals in the Irish aquatic environment and their potential to bioaccumulate in aquatic organisms and pose a risk to human health via dietary intake. Pharmaceutical residues were determined in liquid matrices, such as wastewater effluent, marine surface water (MSW) and artificial seawater (ASW), using solid phase extraction (SPE) in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Pharmaceutical extraction from marine mussels and fish tissues required an additional pressurised liquid extraction (PLE) step prior to SPE and LC-MS/MS. The results of an in situ study, in which caged Mytilus spp. were deployed at three sites on the Irish coastline over a 1-year period are presented. All of the selected pharmaceuticals were quantified in wastewater effluent and marine surface waters and three out of the five monitored pharmaceuticals were detected in environmentally exposed mussel tissue. The potential for pharmaceutical bioaccumulation in fish via trophic level transfer was investigated. A 28-day in vivo experiment was carried out in a flow-through system in which rainbow trout were fed wild marine mussels sampled from one of the most contaminated sites in Ireland. Although low-level residues of trimethoprim were detected in the mussel tissues, no bioaccumulation was reported for this drug or any of the other selected compounds in the liver of the exposed fish. The effect of steaming on the concentrations of five pharmaceutical residues in exposed mussel tissue was investigated in an attempt to assess the potential risk of exposure to humans via ingestion of contaminated seafood. An in vivo experiment was carried out exposing marine mussels to pharmaceutical concentrations via direct injection and water uptake. A selection of water-exposed mussels were cooked (via steaming) resulting in a significant increase of parent pharmaceutical concentrations in the bivalves

Assessing the reliability of uptake and elimination kinetics modelling approaches for estimating bioconcentration factors in the freshwater invertebrate, Gammarus pulex

This study considers whether the current standard toxicokinetic methods are an accurate and applicable assessment of xenobiotic exposure in an aquatic freshwater invertebrate. An in vivo exposure examined the uptake and elimination kinetics for eight pharmaceutical compounds in the amphipod crustacean, Gammarus pulex by measuring their concentrations in both biological material and in the exposure medium over a 96 h period. Selected pharmaceuticals included two anti-inflammatories (diclofenac and ibuprofen), two beta-blockers (propranolol and metoprolol), an anti-depressant (imipramine), an anti-histamine (ranitidine) and two beta-agonists (formoterol and terbutaline). Kinetic bioconcentration factors (BCFs) for the selected pharmaceuticals were derived from a first-order one-compartment model using either the simultaneous or sequential modelling methods. Using the simultaneous method for parameter estimation, BCF values ranged from 12 to 212. In contrast, the sequential method for parameter estimation resulted in bioconcentration factors ranging from 19 to 4533. Observed toxicokinetic plots showed statistically significant lack-of-fits and further interrogation of the models revealed a decreasing trend in the uptake rate constant over time for rantidine, diclofenac, imipramine, metoprolol, formoterol and terbutaline. Previous published toxicokinetic data for 14 organic micro-pollutants were also assessed and similar trends were identified to those observed in this study. The decreasing trend of the uptake rate constant over time highlights the need to interpret modelled data more comprehensively to ensure uncertainties associated with uptake and elimination parameters for determining bioconcentration factors are minimised

Bioaccumulation of metals in juvenile rainbow trout 1 (oncorhynchus mykiss) via dietary exposure to blue mussels

The potential for metals to bioaccumulate in aquatic species, such as fish, via trophic level transfer was investigated. An in vivo experiment was set up in a flow-through system in which juvenile rainbow trout were fed blue mussels collected from a Class A pristine site and an effluent-impacted river estuary, over a period of 28 days. Selected elements (As, Cd, Cr, Co, Cu, Fe, Pb, Mn, Mo, Ni, Se, Sn, V, Zn) were determined in the mussels and fish tissues (muscle and skin) collected at 0, 14 and 28 days. This study reveals the occurrence of metals in mussels sampled in the Irish marine environment and highlights the bioaccumulation potential of metals in fish tissues via trophic transfer. All 14 monitored metals were determined in the mussels collected from both sites and mussels collected from the effluent-impacted site contained three times more Co, Mo, Sn and V than the mussels collected from the Class A site. Following a 28-day dietary exposure, concentrations of As and Se (fish muscle), and Pb, Se and Zn (fish skin), were significantly greater in fish feeding on contaminated mussels compared to those with a regular fish feed diet. The significance of metal detection and bioaccumulation in the mussel and fish tissues, highlights the potential for metal exposure to humans through the food chain. As fish are recommended as a healthy and nutritious food source, it is important to fully understand metal bioaccumulation in commercially important aquatic species and ensure the safety of human consumers

Gradient liquid chromatographic retention time prediction for suspect screening applications: a critical assessment of a generalised artificial neural network-based approach across 10 multi-residue reversed-phase analytical methods

For the first time, the performance of a generalised artificial neural network (ANN) approach for the prediction of 2492 chromatographic retention times (tR) is presented for a total of 1117 chemically diverse compounds present in a range of complex matrices and across 10 gradient reversed-phase liquid chromatography-(high resolution) mass spectrometry methods. Probabilistic, generalised regression, radial basis function as well as 2- and 3-layer multilayer perceptron-type neural networks were investigated to determine the most robust and accurate model for this purpose. Multi-layer perceptrons most frequently yielded the best correlations in 8 out of 10 methods. Averaged correlations of predicted versus measured tR across all methods were R2=0.918, 0.924 and 0.898 for the training, verification and test sets respectively. Predictions of blind test compounds (n=8–84 cases) resulted in an average absolute accuracy of 1.02±0.54 min for all methods. Within this variation, absolute accuracy was observed to marginally improve for shorter runtimes, but was found to be relatively consistent with respect to analyte retention ranges (~5%). Finally, optimised and replicated network dependency on molecular descriptor data is presented and critically discussed across all methods. Overall, ANNs were considered especially suitable for suspects screening applications and could potentially be utilised in bracketed-type analyses in combination with high resolution mass spectrometry

Suspect screening and quantification of trace organic explosives in wastewater using solid phase extraction and liquid chromatography-high resolution accurate mass spectrometry

The first comprehensive assessment of 34 solid phase extraction sorbents is presented for organic explosive residues in wastewater prior to analysis with liquid chromatography-high resolution accurate mass spectrometry (LC-HRMS). A total of 18 explosives were selected including nitramines, nitrate esters, nitroaromatics and organic peroxides. Three polymeric divinylbenzene-based sorbents were found to be most suitable and one co-polymerised with n-vinyl pyrrolidone offered satisfactory recoveries for 14 compounds in fortified wastewater (77–124%). Limits of detection in matrix ranged from 0.026–23 μg L−1 with R2 ≥ 0.98 for most compounds. The method was applied to eight 24-h composite wastewater samples from a London wastewater works and one compound, 2,4-dinitrotoluene, was determined over five days between 332 and 468 g day−1 (225–303 ng L−1). To further exploit the suspect screening capability, 17 additional explosives, precursors and transformation products were screened in spiked wastewater samples. Of these, 14 were detected with recoveries from 62 to 92%, highlighting the broad applicability of the method. To our knowledge, this represents the first screen of explosives-related compounds in wastewater from a major European city. This method also allows post-analysis detection of new or emerging compounds using full-scan HRMS datasets to potentially identify and locate illegal manufacture of explosives via wastewater analysis

The determination of pharmaceutical residues in cooked and uncooked marine bivalves using pressurised liquid extraction, solid-phase extraction and liquid chromatography-tandem mass spectrometry

An optimised and validated method for the determination of pharmaceutical residues in blue mussels (Mytilus spp.) is presented herein, as well as an investigation of the effect of cooking (by steaming) on any potential difference in human exposure risk. Selected pharmaceuticals included two non-steroidal anti-inflammatory drugs (diclofenac and mefenamic acid), an anti-biotic (trimethoprim), an anti-epileptic (carbamazepine) and a lipid regulator (gemfibrozil). An in vivo exposure experiment was set up in the laboratory in which mussels were exposed either directly by injection (10 ng) or daily through spiked artificial seawater (ASW) over 96 h. In liquid matrices, pharmaceutical residues were either determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS) directly, or in combination with solid phase extraction (SPE) for analyte concentration purposes. The extraction of pharmaceuticals from mussel tissues used an additional pressurised liquid extraction (PLE) step prior to SPE and LC-MS/MS. Limits of quantification of between 2 and 46 ng.L-1 were achieved for extracted cooking water and ASW, between 2 and 64 µg.L-1 for ASW in exposure tanks, and between 4 and 29 ng.g-1 for mussel tissue. Method linearities were achieved for pharmaceuticals in each matrix with correlation coefficients of R2>0.975. A selection of exposed mussels was also cooked (via steaming) and analysed using the optimised method to observe any effect on detectable concentrations of parent pharmaceuticals present. An overall increase in pharmaceutical residues in the contaminated mussel tissue and cooking water was observed after cooking

A year-long study of the spatial occurrence and relative distribution of pharmaceutical residues in effluent, receiving marine waters and marine bivalves

Reports concerning the quantitative analysis of pharmaceuticals in marine ecosystems are somewhat limited. It is necessary to determine pharmaceutical fate and assess any potential risk of exposure to aquatic species and ultimately, seafood consumers. In the work presented herein, analytical methods were optimised and validated for the quantification of pharmaceutical residues in wastewater effluent, receiving marine waters and marine mussels (Mytilus spp.). Selected pharmaceuticals included two non-steroidal anti-inflammatory drugs (NSAIDs) (diclofenac and mefenamic acid), an antibiotic (trimethoprim), an antiepileptic (carbamazepine) and a lipid regulator (gemfibrozil). This paper also presents the results of an in situ study in which caged Mytilus spp. were deployed at three sites on the Irish coastline over a 1-year period. In water samples, pharmaceutical residues were determined using solid phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The extraction of pharmaceuticals from mussel tissues used an additional pressurised liquid extraction (PLE) step prior to SPE and LC-MS/MS. Limits of quantification between 15 and 225 ng.L-1 were achieved in wastewater effluent, between 3 and 38 ng.L-1 in marine surface water and between 4 and 29 ng.g-1 dry weight in marine mussels. Method linearity was achieved for pharmaceuticals in each matrix with correlation coefficients of R2>0.976. All five selected pharmaceuticals were quantified in wastewater effluent and marine surface waters. This work has demonstrated the susceptibility of the Mytilus spp. to pharmaceutical exposure following the detection of pharmaceutical residues in the tissues of this mussel species at measurable concentrations

A proteomic evaluation of the effects of the pharmaceuticals diclofenac and gemfibrozil on marine mussels (Mytilus spp.):evidence for chronic sublethal effects on stress- response proteins

Human pharmaceuticals (e.g. the lipid regulator gemfibrozil and the non-steroidal anti-inflammatory drug diclofenac) are an emerging environmental threat in the aquatic environment. This study aimed to evaluate sublethal effects of these two commonly found pharmaceuticals on the protein profiles of marine mussels (Mytilus spp.). Mytilus spp. was exposed to environmentally relevant and elevated concentrations (1 and 1000 mg/l respectively) of both drugs for 14 days. In addition, mussels were maintained for seven days post treatment to examine the potential of blue mussels to recover from such an exposure. Differential protein expression signatures (PES) in the digestive gland of mussels were obtained using two-dimensional gel electrophoresis after 7, 14, and 21 days of exposure. Twelve spots were significantly increased or decreased by gemfibrozil and/or diclofenac, seven of which were successfully identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. These proteins were involved in energy metabolism, oxidative stress response, protein folding, and immune responses. Changes in the PES over time suggested that mussels were still experiencing oxidative stress for up to seven days post exposure. In addition, a suite of biomarkers comprising glutathione transferase, lipid peroxidation, and DNA damage were studied. An oxidative stress response was confirmed by biomarker responses. To our knowledge, this is the first investigation using proteomics to assess the potential effects of human pharmaceuticals on a non-target species in an environmentally-relevant model. The successful application of this proteomic approach supports its potential use in pollution biomonitoring and highlights its ability to aid in the discovery of new biomarkers

Pharmaceuticals in the freshwater invertebrate, Gammarus pulex, determined using pulverised liquid extraction, solid phase extraction and liquid chromatography-tandem mass spectrometry

The development, characterisation and application of a new analytical method for multi-residue PPCP determination in the freshwater amphipod, Gammarus pulex are presented. Analysis was performed using pulverised liquid extraction (PuLE), solid phase extraction (SPE) and liquid chromatography–tandem mass spectrometry (LC–MS/MS). Qualitative method performance offered excellent limits of detection at < 20 ng g− 1 for 18 out of 29 compounds. For quantitative application, linearity and precision were considered acceptable for 10 compounds across the ng-μg g− 1 range (R2 ≥ 0.99; ≤ 20% relative standard deviation respectively). The method was applied to the analysis of G. pulex and river water sourced from six tributaries of the River Thames. Carbamazepine, diazepam, nimesulide, trimethoprim and warfarin were determined in G. pulex samples at low ng g− 1 (dry weight) concentrations across these sites. Temazepam and diclofenac were also detected, but were not quantifiable. Six pharmaceuticals were quantified in surface waters across the eight sites at concentrations ranging from 3 to 344 ng L− 1. The possibility for confirmatory detection and subsequent quantification of pharmaceutical residues in benthic organisms such as G. pulex will enable further understanding on the susceptibility and ecological effects of PPCPs in the aquatic environment

Sorbent film-coated passive samplers for explosives vapour detection Part A

A new thin-film passive sampler is presented as a low resource dependent and discrete continuous monitoring solution for explosives-related vapours. Using 15 mid-high vapour pressure explosives-related compounds as probes, combinations of four thermally stable substrates and six film-based sorbents were evaluated. Meta-aramid and phenylene oxide-based materials showed the best recoveries from small voids (~70%). Analysis was performed using liquid chromatography-high resolution accurate mass spectrometry which also enabled tentative identification of new targets from the acquired data. Preliminary uptake kinetics experiments revealed plateau concentrations on the device were reached between 3–5 days. Compounds used in improvised explosive devices, such as triacetone triperoxide, were detected within 1 hour and were stably retained by the sampler for up to 7 days. Sampler performance was consistent for 22 months after manufacture. Lastly, its direct integration with currently in-service explosives screening equipment including ion mobility spectrometry and thermal desorption mass spectrometry is presented. Following exposure to several open environments and targeted interferences, sampler performance was subsequently assessed and potential interferences identified. High-security building and area monitoring for concealed explosives using such cost-effective and discrete passive samplers can add extra assurance to search routines while minimising any additional burden on personnel or everyday site operation