Multi-residue analysis of 44 pharmaceutical compounds in environmental water samples by solid-phase extraction coupled to liquid chromatography-tandem mass spectrometry.

A solid-phase extraction combined with a liquid chromatography-tandem mass spectrometry analysis has been developed and validated for the simultaneous determination of 44 pharmaceuticals belonging to different therapeutic classes (i.e., antibiotics, anti-inflammatories, cardiovascular agents, hormones, neuroleptics, and anxiolytics) in water samples. The sample preparation was optimized by studying target compounds retrieval after the following processes: i) water filtration, ii) solid phase extraction using Waters Oasis HLB cartridges at various pH, and iii) several evaporation techniques. The method was then validated by the analysis of spiked estuarine waters and wastewaters before and after treatment. Analytical performances were evaluated in terms of linearity, accuracy, precision, detection, and quantification limits. Recoveries of the pharmaceuticals were acceptable, instrumental detection limits varied between 0.001 and 25 pg injected and method quantification limits ranged from 0.01 to 30.3 ng/L. The precision of the method, calculated as relative standard deviation, ranged from 0.3 to 49.4%. This procedure has been successfully applied to the determination of the target analytes in estuarine waters and wastewaters. Eight of these 44 pharmaceuticals were detected in estuarine water, while 26 of them were detected in wastewater effluent. As expected, the highest values of occurrence and concentration were found in wastewater influent.

Determination of Synthetic Musks in Surface Sediment from the Bizerte Lagoon by QuEChERS Extraction Followed by GC-MS.

A new analytical method for the simultaneous determination of eight synthetic musks compounds (SMs) including five polycyclic musks (PCMs) and three nitro musks (NMs) was validated for sediment samples based on a simple QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) sample preparation procedure followed by gas chromatography-mass spectrometry (GC-MS). Good analytical performances were obtained for all the target compounds. For the validation of the method, internal calibration (IC) and internal calibration with QuEChERS (ICQ) were compared. Good linearity was obtained for both calibration methods with determination coefficients (R2) ranging between 0.990 for Musk Xylene (MX) and 0.999 for Tonalide (AHTN) with IC and between 0.991 for Musk Ketone (MK) and 0.999 for Traseolide (ATII) with ICQ. The repeatability ranges were 0.1 %-1.9 % with the IC and 0.1 %-2.6 % with the ICQ. The apparent recoveries obtained for SMs in the standard reference sediment (SRM1944) varied in the range of 70 %-98 % and 75 %-103 % in the sediment from the Bizerte Lagoon (Tunisia). The absolute recoveries ranged between 61 % and 92 % for the SRM1944 and between 61 % and 89 % in the sediment from the Bizerte Lagoon. The limits of detection (LOD) calculated for the two main compounds, Galaxolide (HHCB) and Tonalide (AHTN) were 0.3 and 0.1 ng g-1 respectively. The LODs obtained for ADBI (Celestolide), AHMI (Phantolide), ATII (Traseolide), MM (Muks mosken), MK (Musk Ketone) and MX (Musk Xylene) were 0.08, 0.12, 0.03, 0.34, 0.11, 0.08, 0.10 and 0.15 ng g-1 respectively. The levels of ∑SMs in surface sediments from the Bizerte Lagoon ranged from 1.4 to 4.5 ng g-1, which are 1000 times lower that the predicted no effect concentration (PNEC) for marine organisms.

Assessing and predicting the changes for inorganic mercury and methylmercury concentrations in surface waters of a tidal estuary (Adour Estuary, SW France).

Total and dissolved concentrations of inorganic mercury (IHg) and methylmercury (MeHg) in water (Adour Estuary) were determined during three sampling campaigns and related to biogeochemical variables (nutrients, organic matter). Factors (sampling time, sample type) were included in analysis of covariance with effect separation. The urban estuary suffered historically from anthropogenic sources, however, decreased emissions have reduced Hg concentrations. Total IHg (0.51-3.42 ng L-1) and MeHg (25-81 pg L-1) concentrations are additively described by suspended particulate matter and particulate organic carbon. Higher total concentrations, carried by organic-rich particles, were found near specific discharge points (0.79-8.02 ng L-1 and 34-235 pg L-1 for IHg and MeHg, respectively). The associated high dissolved MeHg concentrations could not be explained only by biogeochemical variables. Better efficiency of the models is found for total than for dissolved concentrations. Models should be checked with other contaminants or with estuaries, suffering from downstream contamination.

Keystone microbial taxa organize micropollutant-related modules shaping the microbial community structure in estuarine sediments.

The fluctuation of environmental conditions drives the structure of microbial communities in estuaries, highly dynamic ecosystems. Microorganisms inhabiting estuarine sediments play a key role in ecosystem functioning. They are well adapted to the changing conditions, also threatened by the presence of pollutants. In order to determine the environmental characteristics driving the organization of the microbial assemblages, we conducted a seasonal survey along the Adour Estuary (Bay of Biscay, France) using 16S rRNA gene Illumina sequencing. Microbial diversity data were combined with a set of chemical analyses targeting metals and pharmaceuticals. Microbial communities were largely dominated by Proteobacteria (41 %) and Bacteroidota (32 %), showing a strong organization according to season, with an important shift in winter. The composition of microbial communities showed spatial distribution according to three main areas (upstream, middle, and downstream estuary) revealing the influence of the Adour River. Further analyses indicated that the microbial community was influenced by biogeochemical parameters (Corg/Norg and δ13C) and micropollutants, including metals (As, Cu, Mn, Sn, Ti, and Zn) and pharmaceuticals (norfloxacin, oxolinic acid and trimethoprim). Network analysis revealed specific modules, organized around keystone taxa, linked to a pollutant type, providing information of paramount importance to understand the microbial ecology in estuarine ecosystems.

Simultaneous determination of 42 pharmaceuticals in seafood samples by solvent extraction coupled to liquid chromatography-tandem mass spectrometry.

An efficient and sensitive analytical method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed and validated for the simultaneous determination of 42 pharmaceuticals belonging to different therapeutic classes (i.e. antibiotics, analgesics, anti-inflammatories, cardiovascular agents, anxiolytics and human indicators) in seafood samples. The very simple sample preparation included analytes extraction with acidified methanol, concentration by evaporation and filtration of the final extract prior to LC-MS/MS analysis. Analytical performances were evaluated in muscles of four commercial species (hake, red mullet, sole and shrimp) and showed good recoveries at two spiked concentration levels, with relative standard deviations below 45%. Limits of quantification ranged from 0.1 to 40.2 ng/g. This procedure has been successfully applied to the determination of the target analytes in seafood collected from the Bay of Biscay (Southern France) and 4 of these 42 pharmaceuticals were detected at low ng/g levels, suggesting a very limited contamination.

Priority and emerging micropollutants distribution from coastal to continental slope sediments: A case study of Capbreton Submarine Canyon (North Atlantic Ocean).

Incising continental margins, submarine canyons are key issue for understanding shelf/deep sea exchange of particulate pollutant, impact on marine ecosystem and global geochemical cycling. The occurrence and distribution of 100 priority and emerging micropollutants were investigated in sediments within the first 25 km of the Capbreton submarine area. The most predominant compounds were polycyclic aromatic hydrocarbons (PAHs), trace metals and metalloid (TMs) (e.g. mercury, lead and arsenic), synthetical musks (e.g. musk ketone, galaxolide), UV filters (e.g. octocrylene and 2-ethylhexyl 4-methoxycinnamate, EHMC) as well as some pharmaceuticals (e.g. azithromycin, acetaminophen). Highest concentrations were measured in submarine canyon sediments, distant from the coast and were correlated with both organic carbon and fine fraction contents, where PAHs, EHMC and musk ketone concentrations up to 7116, 32 and 7 ng g-1 dry weight, respectively. Those results likely demonstrate, that atmospheric inputs of pyrogenic PAHs, and both trapping and transporting of polluted particles along the continuum shore/deep sea by the Capbreton Canyon, might lead to an accumulation of anthropogenic micropollutants. The ecological risk assessment indicates that priority pollutants raise a potentially high risk for benthic organisms (e.g. PAHs, TMs). This might raised a specific concern about how the human can impact this ecosystem.

Microplastics in a salt-wedge estuary: Vertical structure and tidal dynamics.

The abundance and distribution of microplastics in estuaries have been barely documented, and generally without accounting for the vertical structure in the water column. This study presents the very first data on the occurrence and distribution of microplastics in the Adour Estuary, SW France. The experimental data set was complemented by numerical simulations to gain understanding of the behaviour of suspended microplastics. Microplastics were found throughout the water column with a mean abundance of 1.13 part/m3. Films and fragments were the most abundant types of particles collected. Numerical simulations demonstrated that vertical distribution of microplastics in the water column is highly dependent on particle characteristics and on the local hydrodynamics. The main trend is that neutrally-buoyant microplastics are easily flushed out while heavier microplastics are prone to entrapment in the estuary, in particular under low discharge conditions. The present study suggest that estuaries could be a sink of microplastics.

A first estimation of uncertainties related to microplastic sampling in rivers.

Many studies have been conducted to quantify microplastic contamination, but only a few of them have actually the sampling methodology and associated uncertainties. This study seeks to examine the influence of sampling strategy on the confidence interval of river microplastic estimates. 16 samples are collected in the Gave de Pau River (southwestern France) during a three-hour window with a 330-µm mesh size net. Three different exposure times (3, 5 and 7 min) allow for a respective filtration rate by the net of 35.6 m3 (3 samples), 59.4 m3 (10 samples), and 83.2 m3 (3 samples) of water. Organic matter contained in samples is removed by hydrogen peroxide oxidation. The plastic particles are then counted and classified under a binocular microscope. The microplastic concentrations vary between 2.64 and 4.24 microplastics/m3, with a median value of 3.26 microplastics/m3. Statistical analysis does not show differences in microplastic concentrations for the three exposure times. This result seems to demonstrate that a filtration of approx. 35 m3 of water is sufficient under similar conditions (similar flow condition and degree of microplastic contamination) and can help reduce sampling and sample processing time. Other analyses, based on 10 filtrations of 59.4 m3, show that the higher the number of samples, the lower the confidence interval. For triplicates, the mean confidence interval reaches 15% of the median value. Thus, collecting triplicates would seem to offer a reasonable optimum, in combining an acceptable error percentage and time efficiency. These results might depend on the microplastic load of the river, therefore making it necessary to conduct similar analyses on other rivers. This study reports for the first time uncertainties related to microplastic sampling in rivers. Such findings will serve to set up long term monitoring, highlight spatial differences between sites and improve the accuracy of annual microplastic fluxes in rivers.

Inputs, dynamics and potential impacts of silver (Ag) from urban wastewater to a highly turbid estuary (SW France).

Although silver (Ag) has been listed as a priority pollutant for the aquatic environment by the European Union (Directive 2006/11/EC), the use of Ag-based products with antimicrobial effects is increasing in Europe, as well as North America and Asia. This study investigates personal care products (PCP) as a potential source of Ag in wastewater, as well as the dynamics and fate of Ag in the influent and effluent of a major urban wastewater treatment plant (WWTP) located on the fluvial part of the Gironde Estuary. Typical household PCPs marked as using Ag contained concentrations of up to 0.4 mg kg-1 making them likely contributors to urban Ag released into the aquatic environment. Silver concentrations in influent wastewater generally occurred during mid-week working hours and decreased during the night and on weekends clearly indicating the dominance of urban sources. Up to 90% of the total Ag in wastewater was bound to particles and efficiently (>80%) removed by the treatment process, whereas 20% of Ag was released into the fluvial estuary. Silver concentrations in wastewater effluents clearly exceeded estuarine concentrations and may strongly amplify the local Ag concentrations and fluxes, especially during summer rainstorms in low river discharge conditions. Further work should focus on environmental effects and fate of urban Ag release due to immediate localized outfall and/or the adsorption on estuarine particles and subsequent release as dissolved Ag chloro-complexes within the estuarine salinity gradient.

Assessment of the effects of Cu and Ag in oysters Crassostrea gigas (Thunberg, 1793) using a battery of cell and tissue level biomarkers.

Oysters are considered sentinel organisms in environmental water quality monitoring programs in which cell and tissue level biomarkers are reliable tools. Copper (Cu) and silver (Ag) are present in relatively high concentrations in several estuaries, potentially affecting environmental and human health. Crassostrea gigas oysters were exposed during 28 days to a range of environmentally relevant concentrations of Cu and Ag alone or in mixture. Effects were studied through cell and tissue level biomarkers approach. Results indicated: changes in the Condition Index (CI), altered digestive gland epithelium and presence of histopathological alterations in the gonad and digestive gland of exposed oysters. A time-dependent increase in lipofuscin contents in exposed oysters and an increase in intralysosomal metal accumulation in digestive cells through the experiment were also recorded. The Integrative Biological Response (IBR) Index showed that even at low exposure levels, Ag and Cu can produce alterations on oysters' health status.

Relationships between bacterial energetic metabolism, mercury methylation potential, and hgcA/hgcB gene expression in Desulfovibrio dechloroacetivorans BerOc1.

The proteins encoded by the hgcA and hgcB genes are currently the only ones known to be involved in the mercury methylation by anaerobic microorganisms. However, no studies have been published to determine the relationships between their expression level and the net/gross methylmercury production. This study aimed to decipher the effect of growth conditions on methylmercury production and the relationships between hgcA and hgcB expression levels and net methylation. Desulfovibrio dechloroacetivorans strain BerOc1 was grown under sulfidogenic conditions with different carbon sources and electron donors as well as under fumarate respiration. A good correlation was found between the biomass production and the methylmercury production when the strain was grown under sulfate-reducing conditions. Methylmercury production was much higher under fumarate respiration when no sulfide was produced. During exponential growth, hgcA and hgcB gene expression levels were only slightly higher in the presence of inorganic mercury, and it was difficult to conclude whether there was a significant induction of hgcA and hgcB genes by inorganic mercury. Besides, no relationships between hgcA and hgcB expression levels and net mercury methylation could be observed when the strain was grown either under sulfate reduction or fumarate respiration, indicating that environmental factors had more influence than expression levels.

Mercury mobilization and speciation linked to bacterial iron oxide and sulfate reduction: A column study to mimic reactive transfer in an anoxic aquifer.

Mercury (Hg) mobility and speciation in subsurface aquifers is directly linked to its surrounding geochemical and microbial environment. The role of bacteria on Hg speciation (i.e., methylation, demethylation and reduction) is well documented, however little data is available on their impact on Hg mobility. The aim of this study was to test if (i) Hg mobility is due to either direct iron oxide reduction by iron reducing bacteria (IRB) or indirect iron reduction by sulfide produced by sulfate reducing bacteria (SRB), and (ii) to investigate its subsequent fate and speciation. Experiments were carried out in an original column setup combining geochemical and microbiological approaches that mimic an aquifer including an interface of iron-rich and iron depleted zones. Two identical glass columns containing iron oxides spiked with Hg(II) were submitted to (i) direct iron reduction by IRB and (ii) to indirect iron reduction by sulfides produced by SRB. Results show that in both columns Hg was leached and methylated during the height of bacterial activity. In the column where IRB are dominant, Hg methylation and leaching from the column was directly correlated to bacterial iron reduction (i.e., Fe(II) release). In opposition, when SRB are dominant, produced sulfide induced indirect iron oxide reduction and rapid adsorption of leached Hg (or produced methylmercury) on neoformed iron sulfides (e.g., Mackinawite) or its precipitation as HgS. At the end of the SRB column experiment, when iron-oxide reduction was complete, filtered Hg and Fe concentrations increased at the outlet suggesting a leaching of Hg bound to FeS colloids that may be a dominant mechanism of Hg transport in aquifer environments. These experimental results highlight different biogeochemical mechanisms that can occur in stratified sub-surface aquifers where bacterial activities play a major role on Hg mobility and changes in speciation.

Sources and historical record of tin and butyl-tin species in a Mediterranean bay (Toulon Bay, France).

Concentrations of inorganic tin (Sn(inorg)), tributyltin (TBT) and its degradation products dibutyltin (DBT) and monobutyltin (MBT) were measured in surface sediments and in two cores from the Toulon Bay, hosting the major French military harbour. Anticipating planned dredging, the aim of the present work is to map and evaluate for the first time the recent and historic contamination of these sediments by inorganic and organic Sn species derived from antifouling paints used for various naval domains including military, trade, tourism and leisure. Tin and butyl-Sn concentrations in the bay varied strongly (4 orders of magnitude), depending on the site, showing maximum values near the shipyards. The concentrations of total Sn (1.3-112 µg g(-1)), TBT (<0.5-2,700 ng g(-1)), DBT (<0.5-1,800 ng g(-1)) and MBT (0.5-1,000 ng g(-1)) generally decreased towards the open sea, i.e. as a function of both distance from the presumed main source and bottom currents. Progressive degradation state of the butyl-Sn species according to the same spatial scheme and the enrichment factors support the scenario of a strongly polluted bay with exportation of polluted sediment to the open Mediterranean. Low degradation and the historical records of butyl-Sn species in two (210)Pb-dated sediment cores, representative of the Northern Bay, are consistent with the relatively recent use of TBT by military shipyards and confirm maximum pollution during the 1970s, which will persist in the anoxic sediments for several centuries. The results show that (a) degradation kinetics of butyl-Sn species depend on environmental conditions, (b) the final degradation product Sn(inorgBT) is by far the dominant species after 10-12 half-life periods and (c) using recent data to reliably assess former TBT contamination requires the use of a modified butyl-Sn degradation index BDI(mod). Resuspension of extremely contaminated subsurface sediments by the scheduled dredging will probably result in mobilization of important amounts of butyl-Sn species.

Silver behaviour along the salinity gradient of the Gironde Estuary.

Dissolved and particulate Ag concentrations (Ag(D) and Ag(P), respectively) were measured in surface water and suspended particulate matter (SPM) along the salinity gradient of the Gironde Estuary, South West France, during three cruises (2008-2009) covering contrasting hydrological conditions, i.e. two cruises during intermediate and one during high freshwater discharge (~740 and ~2,300 m(3)/s). Silver distribution reflected non-conservative behaviour with 60-70 % of Ag(P) in freshwater particles being desorbed by chlorocomplexation. The amount of Ag(P) desorbed was similar to the so-called reactive, potentially bioavailable Ag(P) fraction (60 ± 4 %) extracted from river SPM by 1 M HCl. Both Ag(P) (0.22 ± 0.05 mg/kg) and Ag(P)/Th(P) (0.025-0.028) in the residual fraction of fluvial and estuarine SPM were similar to those in SPM from the estuary mouth and in coastal sediments from the shelf off the Gironde Estuary, indicating that chlorocomplexation desorbs the reactive Ag(P). The data show that desorption of reactive Ag(P) mainly occurs inside the estuary during low and intermediate discharge, whereas expulsion of partially Ag(P)-depleted SPM (Ag(P)/Th(P) ~0.040) during the flood implies ongoing desorption in the coastal ocean, e.g. in the nearby oyster production areas (Marennes-Oléron Bay). The highest Ag(D) levels (6-8 ng/L) occurred in the mid-salinity range (15-20) of the Gironde Estuary and were decoupled from freshwater discharge. In the maximum turbidity zone, Ag(D) were at minimum, showing that high SPM concentrations (a) induce Ag(D) adsorption in estuarine freshwater and (b) counterbalance Ag(P) desorption in the low salinity range (1-3). Accordingly, Ag behaviour in turbid estuaries appears to be controlled by the balance between salinity and SPM levels. The first estimates of daily Ag(D) net fluxes for the Gironde Estuary (Boyle's method) showed relatively stable theoretical Ag(D) at zero salinity (Ag (D) (0) = 25-30 ng/L) for the contrasting hydrological situations. Accordingly, Ag(D) net fluxes were very similar for the situations with intermediate discharge (1.7 and 1.6 g/day) and clearly higher during the flood (5.0 g/day) despite incomplete desorption. Applying Ag (D) (0) to the annual freshwater inputs provided an annual net Ag(D) flux (0.64-0.89 t/year in 2008 and 0.56-0.77 t/year in 2009) that was 12-50 times greater than the Ag(D) gross flux. This estimate was consistent with net Ag(D) flux estimates obtained from gross Ag(P) flux considering 60 % desorption in the estuarine salinity gradient.

Long-term records of cadmium and silver contamination in sediments and oysters from the Gironde fluvial-estuarine continuum - evidence of changing silver sources.

The Gironde fluvial estuarine system is impacted by historic metal pollution (e.g. Cd, Zn, Hg) and oysters (Crassostrea gigas) from the estuary mouth have shown extremely high Cd concentrations for decades. Based on recent work (Chiffoleau et al., 2005) revealing anomalously high Ag concentrations (up to 65 mg kg(-1); dry weight) in Gironde oysters, we compared long-term (~1955-2001) records of Ag and Cd concentrations in reservoir sediment with the respective concentrations in oysters collected between 1979 and 2010 to identify the origin and historical trend of the recently discovered Ag anomaly. Sediment cores from two reservoirs upstream and downstream from the main metal pollution source provided information on (i) geochemical background (upstream; Ag: ~0.3 mg kg(-1); Cd: ~0.8 mg kg(-1)) and (ii) historical trends in Ag and Cd pollution. The results showed parallel concentration-depth profiles of Ag and Cd supporting a common source and transport. Decreasing concentrations since 1986 (Cd: from 300 to 11 mg kg(-1); Ag: from 6.7 to 0.43 mg kg(-1)) reflected the termination of Zn ore treatment in the Decazeville basin followed by remediation actions. Accordingly, Cd concentrations in oysters decreased after 1988 (from 109 to 26 mg kg(-1), dry weight (dw)), while Ag bioaccumulation increased from 38 up to 116 mg kg(-1), dw after 1993. Based on the Cd/Ag ratio (Cd/Ag~2) in oysters sampled before the termination of zinc ore treatment (1981-1985) and assuming that nearly all Cd in oysters originated from the metal point source, we estimated the respective contribution of Ag from this source to Ag concentrations in oysters. The evolution over the past 30 years clearly suggested that the recent, unexplained Ag concentrations in oysters are due to increasing contributions (>70% after 1999) by other sources, such as photography, electronics and emerging Ag applications/materials.