Critical evaluation of the potential of ICP-MS-based systems in toxicological studies of metallic nanoparticles.

The extensive application of metallic nanoparticles (NPs) in several fields has significantly impacted our daily lives. Nonetheless, uncertainties persist regarding the toxicity and potential risks associated with the vast number of NPs entering the environment and human bodies, so the performance of toxicological studies are highly demanded. While traditional assays focus primarily on the effects, the comprehension of the underlying processes requires innovative analytical approaches that can detect, characterize, and quantify NPs in complex biological matrices. Among the available alternatives to achieve this information, mass spectrometry, and more concretely, inductively coupled plasma mass spectrometry (ICP-MS), has emerged as an appealing option. This work critically reviews the valuable contribution of ICP-MS-based techniques to investigate NP toxicity and their transformations during in vitro and in vivo toxicological assays. Various ICP-MS modalities, such as total elemental analysis, single particle or single-cell modes, and coupling with separation techniques, as well as the potential of laser ablation as a spatially resolved sample introduction approach, are explored and discussed. Moreover, this review addresses limitations, novel trends, and perspectives in the field of nanotoxicology, particularly concerning NP internalization and pathways. These processes encompass cellular uptake and quantification, localization, translocation to other cell compartments, and biological transformations. By leveraging the capabilities of ICP-MS, researchers can gain deeper insights into the behaviour and effects of NPs, which can pave the way for safer and more responsible use of these materials.

Critical evaluation of key parameters in single particle ICP-MS data processing for the correct determination of platinum nanoparticles in complex environmental and biological matrices.

There is an urgent need for the harmonization of critical parameters in single particle inductively coupled plasma mass spectrometry (SP-ICP-MS) and they have been deeply studied and optimized in the present work using platinum nanoparticles (PtNPs) as a representative case of study. Special attention has been paid to data processing in order to achieve an adequate discrimination between signals. Thus, a comparison between four different algorithms has been performed and the method for transport efficiency calculation has also been thorougly evaluated (finding the use of a well-characterized solution of the same targeted analyte (30 nm PtNPs) as adequate). The best results have been obtained after the application of a deconvolution approach for the data processing and using 5 ms as dwell time and 40,000 data points for data acquisition. Under the optimized conditions, a correct discrimination between NP events and background signal up to 100 or 750 ng L-1 of added ionic Pt was reached for 30 and 50 nm PtNPs, respectively. The suitability of the developed method for the characterization of PtNPs in relevant environmental (water samples) and biological (cell culture media) matrices has also been demonstrated.

Hair mercury is associated with dyslipidemia and cardiovascular risk: An anthropometric, biochemical and genetic cross-sectional study of Amazonian vulnerable populations.

This cross-sectional study evaluated the association between human exposure to mercury and cardiovascular risk using lipid profile (including apolipoproteins) and genetic analysis of Amazonian riverine population. Anthropometric data (gender, age, height, weight, blood pressure, and neck and waist circumferences) of the participants were recorded. Total mercury and methylmercury (MeHg) content were quantified in hair by ICP-MS and GC-pyro-AFS system. Polymorphisms rs662799, rs693, rs429358 and rs7412 (of genes of apolipoproteins A-V, B, and E at positions 112 and 158, respectively) were genotyped by real-time PCR. The population presented a dyslipidemia profile significantly correlated with high mercury levels. The apolipoprotein B/apolipoprotein A-I (ApoB/ApoA-I) index was also positively correlated with mercury, supporting a possible causal relationship. Allelic distributions were similar to those described in other populations, suggesting that genetic susceptibility may not have a significant role in the lipid alterations found in this work. This study demonstrated for the first time: i) the relationship between mercury exposure and cardiovascular risk-related apolipoproteins in humans, ii) the ApoB levels and the ApoB/ApoA-I index as the risk factors more strongly associated to the mercury-related dyslipidemia in humans, and iii) the prevalence of high/moderate risk of acute myocardial infarction in the vulnerable and chronically exposed-populations of the Amazon, in addition to the genotypic profile of the three most frequent polymorphisms in apolipoproteins of relevance for cardiovascular risk. This early detection of lipid alterations is essential to prevent the development of cardiovascular diseases (CVD), especially in chronically exposed populations such as those found in the Amazon. Therefore, in addition to provide data for the Minamata Convention implementation, our work is in line with the efforts joined by all members of the World Health Organization committed to reducing premature deaths originating from non-communicable diseases by 25% in 2025, including CVD.

Electrical asymmetric-flow field-flow fractionation with a multi-detector array platform for the characterization of metallic nanoparticles with different coatings.

Electrical asymmetric-flow field-flow fractionation (EAF4) is a new and interesting analytical technique recently proposed for the characterization of metallic nanoparticles (NPs). It has the potential to simultaneously provide relevant information about size and electrical parameters, such as electrophoretic mobility (µ) and zeta-potential (ζ), of individual NP populations in an online instrumental setup with an array of detectors. However, several chemical and instrumental conditions involved in this technique are definitely influential, and only few applications have been proposed until now. In the present work, an EAF4 system has been used with different detectors, ultraviolet-visible (UV-vis), multi-angle light scattering (MALS), and inductively coupled plasma with triple quadrupole mass spectrometry (ICP-TQ-MS) for the characterization of gold, silver, and platinum NPs with both citrate and phosphate coatings. The behavior of NPs has been studied in terms of retention time and signal intensity under both positive and negative current with results depending on the coating. Carrier composition, particularly ionic strength, was found to be critical to achieve satisfactory recoveries and a reliable measurement of electrical parameters. Dynamic light scattering (DLS) has been used as a comparative technique for these parameters. The NovaChem surfactant mix (0.01%) showed a quantitative recovery (93 ± 1%) of the membrane, but the carrier had to be modified by increasing the ionic strength with 200 µM of Na2CO3 to achieve consistent µ values. However, ζ was one order of magnitude lower in EAF4-UV-vis-MALS than in DLS, probably due to different electric processes in the channel. From a practical point of view, EAF4 technique is still in its infancy and further studies are necessary for a robust implementation in the characterization of NPs.

Speciation of platinum nanoparticles in different cell culture media by HPLC-ICP-TQ-MS and complementary techniques: A contribution to toxicological assays.

Toxicological studies of nanoparticles (NPs) are highly demanded nowadays but they are very challenging. In the in vitro assays, the understanding of the role of cell culture media is crucial to derive a proper interpretation of the toxicological results and to do so, new analytical tools are necessary. In this context, an analytical strategy based on reversed-phase liquid chromatography hyphenated to inductively coupled plasma-triple quadrupole mass spectrometry (HPLC-ICP-TQ-MS) has been developed for the first time for the detection and characterization of both 5 and 30 nm PtNPs, as well as ionic platinum species, in commonly used cell culture media. For this purpose, Dulbecco's Modified Eagle Medium, DMEM-high glucose, DMEM-F12, DMEM 31053-028, and Roswell Park Memorial Institute, RPMI-1640 (supplemented with 10% fetal bovine serum (FBS) and antibiotics) at several incubation times (24, 48, and 96 h at 37 °C) were tested. After a careful optimization and analytical performance, the developed method allows to simultaneously study the oxidation process, leading to the release of ionic species, and the increase in the hydrodynamic volume of PtNPs, probably related to the formation of new biological entities (protein corona). The magnitude of both processes was found to be dependent on the tested cell culture media and incubation times. Dynamic light scattering (DLS) and high-resolution scanning electron microscopy (HR-SEM) were used as complementary techniques to study the important process of both soft and hard protein corona formation. The feasibility of the HPLC-ICP-TQ-MS to get relevant information for toxicological studies has been demonstrated and in light of our results, the influence of the cell culture media on the behavior of PtNPs should not be underestimated.

Vasovagal syncope after blood donation in active duty military personnel of the Spanish Army.

BACKGROUND: All studies of vasovagal syncope (VVS) after blood donation have been performed with civilian donors. We hypothesized that military donors have a lower incidence of VVS and a particular set of predisposing factors. STUDY DESIGN AND METHODS: Retrospective case-control study matching every case of VVS seen from 2011 to 2019 with two controls without VVS from the same blood drive. We used the odds ratio (OR) and its 95% confidence interval (CI), estimated by multivariate logistic regression, to identify independent predictors of VVS. RESULTS: There were 105 episodes of VVS among 65.481 whole blood donations (0.15%). VVS was more frequent among donors from military academies than from other military units (0.37% vs. 0.10%, p < .001) and in collections conducted in mobile inside setups than in mobile buses (0.23% vs. 0.06%, p < .001). In the multivariate analysis, the only independent predictors of VVS were the status of first-time donor (OR 2.6, 95% CI 1.5-4.4; p < .001) and pre-donation systolic blood pressure (SBP) < 120 mm Hg (OR 1.9, 95% CI 1.1-3.3; p = .01). Donors with both risk factors had a 5.5-fold increased risk of VVS than donors without any risk factor. Age and female sex were not predictive of VVS. DISCUSSION: Active duty military blood donors have a lower incidence of VVS than that reported in civilian donors. First-time donors and donors with SBP < 120 mm Hg should be temporarily deferred when immediate reincorporation to hazardous or strenuous duty tasks after donation is inescapable.

A simple analytical methodology for platinum nanoparticles control in complex clinical matrices via SP-ICP-MS.

A simple method based on the use of inductively coupled plasma mass spectrometry in single particle mode (SP-ICP-MS) has been proposed, for the first time, for the study of platinum nanoparticles (PtNPs) in complex clinical matrices such as human urine and blood serum. Critical parameters for signal acquisition were optimized to achieve a correct and simultaneous sizing and counting (particle-based in particles L-1 and mass-based in ng L-1) of 50 and 70 nm PtNPs. Different reagents, as tetramethylammonium hydroxide (TMAH) and/or Triton X-100, and concentrations have been tested to ensure an adequate stabilization and extraction of PtNPs. Finally, TMAH at 1% is demonstrated to be the best reagent to extract the NPs guaranteeing their integrity. No heating or any additional treatment was required, which allows sample preparation, and the overall process, to be simple and fast. Good precisions for size (2% RSD) and particle number and mass concentrations (<1% RSD), and limits of detection of 21.6 nm and 1.9 × 105 particles L-1 were reported. The influence of matrix on the determination of PtNP sizes and number- and mass-based concentrations was evaluated. Particle sizes were in all cases in accordance with values determined by TEM or SEM, whereas recoveries of PtNPs in terms of concentration ranged between 92 and 101%. The stability of PtNP characteristics after 24 h was specifically studied in human urine spiked with PtNPs. Statistically significant differences were only reported for the particle number concentrations of 50 nm PtNPs in female urine samples. The present work will be relevant to understand the behaviour of PtNPs in body fluids and to take appropriate actions in future (pre)clinical trials.

Nutrient enrichment and trace element accumulation in sediments caused by waterbird colonies at a Mediterranean semiarid floodplain.

Waterfowl are vectors of nutrients, metals, and other pollutants (i.e. PCBs and microorganisms) on wetlands and other water bodies, especially when bird population density is high, e.g., in nesting, roosting or breeding periods. This work reports the effects caused by the increase in breeding pairs of herons and cormorants between 2010 and 2014 on sediments in an eutrophic Mediterranean wetland belonging to La Mancha Húmeda Biosphere Reserve. Nutrients, metals, metalloids, and isotopic composition (δ15N and δ13C) were analysed in sediments and faeces at several points in the Tablas de Daimiel National Park (TDNP), as well as in the nearby Navaseca wetland, which receives inputs from a wastewater treatment plant (WWTP). Sediments from Navaseca and from one site that is affected by heron colonies (Garcera) in the TDNP showed higher concentrations of organic matter, total organic carbon, total nitrogen, total phosphorus, Al, Co, Cu, Cr, Fe, Ni and Zn than the rest of the sampling sites. Significant correlations between these variables were found, probably suggesting that they have a common organic source. The highest δ15N values in sediments of the TDNP were registered in Guadiana, a site where there is no impact from colonies; these values might therefore be due to anthropogenic activities. Metal and metalloid content in sediments was lower than the regional reference values in soils, except for Se in the TDNP, and Zn and Cu in Navaseca. High Se level in TDNP sediment may be caused by seleniferous soils located upstream. Results showed a low impact of bird droppings compared to other sources of nutrients in the annual input in the TDNP.

Metals and metalloids in freshwater fish from the floodplain of Tablas de Daimiel National Park, Spain.

The Tablas de Daimiel National Park (TDNP) is a floodplain ecosystem in central Spain with a potential risk of heavy metal and metalloid pollution. The objective of this study was to know the accumulation of arsenic (As), cadmium (Cd), mercury (Hg), lead (Pb), and selenium (Se) in muscle of six species of freshwater fish from the floodplain of TDNP. We obtained muscle samples of Cyprinus carpio (n = 89); Squalius pyrenaicus (n = 16); Ameiurus melas (n = 9); Lepomis gibbosus (n = 8); Micropterus salmoides (n = 6) and Carassius auratus (n = 5). A. melas, which is a predatory species, had significantly higher Hg concentrations than omnivorous or herbivorous species (i.e. C. carpio). On the contrary, A. melas showed lower concentrations of As, Pb and Se than omnivorous species (i.e. S. pyrenaicus and L. gibbosus ). The concentration of Hg was positively associated with fish size in C. carpio and A. melas. Some individuals of C. carpio (5.7%) and S. pyrenaicus (12.5%) showed Pb muscle concentrations above the maximum residue levels established by the European legislation for human consumers. The observed muscle Se concentrations can be associated with adverse effects on fish such as blood changes, reduced growth, mortality of juveniles and reproductive failure. The accumulation of Se in this floodplain located in a seleniferous area and the contamination produced by spent Pb shot pellets used for hunting in the past are discussed as potential sources of the elevated levels of these two elements in fish from this floodplain wetland.

Dust and bullets: Stable isotopes and GPS tracking disentangle lead sources for a large avian scavenger.

Lead intoxication is an important threat to human health and a large number of wildlife species. Animals are exposed to several sources of lead highlighting hunting ammunition and lead that is bioavailable in topsoil. Disentangling the role of each in lead exposure is an important conservation issue, particularly for species potentially affected by lead poisoning, such as vultures. The identification of lead sources in vultures and other species has been classically addressed by means of stable-isotope comparisons, but the extremely varied isotope signatures found in ammunition hinders this identification when it overlaps with topsoil signatures. In addition, assumptions related to the exposure of individual vultures to lead sources have been made without knowledge of the actual feeding grounds exploited by the birds. Here, we combine lead concentration analysis in blood, novel stable isotope approaches to assign the origin of the lead and GPS tracking data to investigate the main foraging grounds of two Iberian griffon vulture populations (N = 58) whose foraging ranges differ in terms of topsoil lead concentration and intensity of big game hunting activity. We found that the lead signature in vultures was closer to topsoil than to ammunition, but this similarity decreased significantly in the area with higher big game hunting activity. In addition, attending to the individual home ranges of the tracked birds, models accounting for the intensity of hunting activity better explained the higher blood lead concentration in vultures than topsoil exposure. In spite of that, our finding also show that lead exposure from topsoil is more important than previously thought.

Simultaneous extraction and preconcentration of monomethylmercury and inorganic mercury using magnetic cellulose nanoparticles.

Magnetite (Fe3O4) nanoparticles were modified with nanocellulose and are showed to be a useful sorbent for magnetic solid-phase extraction of mercury species. Speciation analysis was performed by using gas chromatography coupled to atomic fluorescence detection (GC-pyro-AFS). The magnetic properties of the sorbent make this approach simple and rapid, and the use of a renewable and biodegradable nanomaterial (nanocellulose) makes it environmentally friendly. The factors that affect adsorption (pH value, amount of nanomaterial, time, volume of sample) and desorption (solvent, time) have been optimized. Both desorption and derivatization of mercury species were performed in a single step. This reduces considerably the sample preparation time. Under the optimized conditions, the limits of detection are 4.0 pg mL-1 for monomethylmercury and 5.6 pg mL-1 for inorganic mercury. The repeatability and reproducibility are satisfactory. The method enables inorganic mercury and monomethylmercury to be simultaneously extracted, with preconcentration factors up to 300. The potential interferences of organic matter and/or co-existing ions were also investigated using synthetic waters. The procedure was applied to the analysis of tap water and river water samples with different characteristics from a mercury polluted area (Almadén, Spain). The extraction recoveries ranged from 81 to 98% regardless of the type of water, which demonstrates the applicability of the method. This is the first time that this kind of sorbent is used for trace metal speciation. Graphical abstract Schematic representation of the new composite material (made of Fe3O4 magnetic nanoparticles and cellulose fibers, MCNPs) for the simultaneous extraction and preconcentration of mercury species taking advantage of the magnetic properties of this eco-friendly sorbent.

Mercury species accumulation and distribution in Typha domingensis under real field conditions (Almadén, Spain).

Monomethylmercury (MeHg) is one of the most toxic and the most commonly occurring organomercury compound and the wetlands are one of the main areas of generation of this Hg form. Concretely, it is in the macrophyte root system where better conditions are given for its generation. However, the knowledge of absorption and subsequent distribution of mercury (Hg) and monomethylmercury in aquatic plants is still limited. Mercury mining district such as Almadén (Ciudad Real, Spain) is a natural laboratory where different rivers flow and the species Typha domingensis Pers. is a common macrophyte which grows in their riverbanks. The aim of the present work is to apply a recently developed method specially designed to analyze Hg species in plant tissues to the different fractions of T. domingensis under real field conditions and to study the accumulation and distribution of Hg species (inorganic Hg and MeHg) within the plant. The results proved that whatever Hg species has preference to be accumulated in the belowground fractions and demonstrated a high efficiency in the accumulation of MeHg.

Metals and metalloids in blood and feathers of common moorhens (Gallinula chloropus) from wetlands that receive treated wastewater.

We addressed the hypothesis that birds in eutrophic wetlands receiving wastewater treatment plant (WWTP) effluents are exposed to high levels of metals and metalloids and this may drive an ecological trap in some species attracted to these highly productive ecosystems. Levels of metals and metalloids were determined in sediment and in blood and feathers of common moorhens (Gallinula chloropus) from two wetlands in Central Spain: Navaseca Pond, which receives directly the effluent of a WWTP; and Tablas de Daimiel National Park, which is a floodplain less affected by urban discharges. Sediment concentrations of Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd and Pb were higher in Navaseca Pond than in Tablas de Daimiel; only Se was higher in Tablas de Daimiel than in Navaseca. Blood levels of Hg and Se were higher in moorhens from Tablas de Daimiel than those from Navaseca. In the case of Hg these levels were below the threshold of adverse effect, but Se levels in 24% of moorhens from Tablas de Daimiel were above the threshold value associated with Se toxicity in birds (1000 ng/mL). In feathers, Hg, Se, Mn, Cu and As levels were higher in Tablas de Daimiel than in Navaseca. Body condition of moorhens was negatively associated with blood Se levels in the moorhens from Tablas de Daimiel. We can reject the hypothesis of a higher accumulation of metals and metalloids in birds associated with the WWTP effluent, but Se levels may need further research considering the nature of the floodplain of Tablas de Daimiel National Park.

Assessing mercury intoxication in isolated/remote populations: Increased S100B mRNA in blood in exposed riverine inhabitants of the Amazon.

Mercury is a heavy metal responsible for human intoxication worldwide and especially in the Amazon, where both natural and anthropogenic sources are responsible for exposure in riverine populations. Methylmercury is the most toxic specie of mercury with recognized neurotoxicity due to its affinity for the central nervous system. S100B protein is a well-established biomarker of brain damage and it was recently associated with mercury-related neurotoxicity. Accurate measurement is especially challenging in isolated/remote populations due to the difficulty of adequate sample conservation, therefore here we use S100B mRNA levels in blood as a way to assay mercury neurotoxicity. We hypothesized that individuals from chronically exposed populations showing mercury levels above the limit of 10 µg/g in hair would present increased levels of S100B mRNA, likely due to early brain damage. A total of 224 riverine individuals were evaluated for anthropometric data (age, body mass index), self-reported symptoms of mercury intoxication, c-reactive protein in blood, and mercury speciation in hair. Approximately 20% of participants showed mercury levels above the limit, and prevalence for most symptoms was not different between individuals exposed to high or low mercury levels. Rigorous exclusion criteria were applied to avoid confounding factors and S100B mRNA in blood was tested by RT-qPCR. Participants with ≥10 µg/g of mercury had S100B mRNA levels over two times higher than that of individuals with lower exposure. A significant correlation was also detected between mercury content in hair and S100B mRNA levels in blood, supporting the use of the latter as a possible candidate to predict mercury-induced neurotoxicity. This is the first report of an association between S100B mRNA and mercury exposure in humans. The combination of both exposure and intoxication biomarkers could provide additional support for the screening and early identification of high-risk individuals in isolated populations and subsequent referral to specialized centers.

Genetic Susceptibility to Neurodegeneration in Amazon: Apolipoprotein E Genotyping in Vulnerable Populations Exposed to Mercury.

Human exposure to mercury is a serious problem of public health in Amazon. As in other vulnerable populations throughout the world, Amazonian riverine populations are chronically exposed to this metal and some symptoms of mercury intoxication were already detected in these populations. However, studies on the genetic susceptibility to mercury toxicity in the Amazon are scarce, and they tested a limited number of individuals. In this context, apolipoprotein E gene (APOE) is a key element with a well-established association among their alleles and the neurodegenerative consequences of mercury intoxication. However, no studies have addressed APOE genotyping in Amazonian exposed populations. Additionally, epidemiological studies with APOE genotyping in Amazon have been restricted to indigenous populations. Therefore, this work analyzed for the first time the genotypic and allelic profiles of APOE in Amazonian riverine populations chronically exposed to mercury. Eight hundred and twenty three individuals were enrolled in our study donating blood (794) and/or hair (757). APOE genotyping was analyzed by real-time PCR. Total mercury and mercury species were quantified by ICP-MS and GC-pyro-AFS, respectively. Genomic ancestry markers were evaluated by multiplex-PCR reaction, separated by capillary electrophoresis on the ABI 3130 Genetic Analyzer instrument and analyzed on GeneMapper ID v3.2. The 𝜀3 and 𝜀3/𝜀3 were the most frequent allele and genotype, respectively, followed by 𝜀4 allele and 𝜀3/𝜀4 genotype. Only 𝜀2/𝜀2 genotype was not found, suggesting that the absence of this genotype is a generalized phenomenon in Amazon. Also, our data supported an association between the presence of APOE4 and the Amerindian origin in these populations. Fifty-nine individuals were identified at maximum risk with levels of mercury above 10 µg/g and the presence of APOE4. Interestingly, among individuals with high mercury content, APOE4-carriers had high mercury levels than APOE2-carriers, pointing to a different heavy metal accumulation according to the APOE allele. These data suggest that APOE4, in addition to a possible pharmacodynamic effect, may influence pharmacokinetically the mercury exposure causing its higher accumulation and leading to worse deleterious consequences. Our results may aid in the development of prevention strategies and health policy decision-making regarding these at-risk vulnerable populations.

Carbon nanotubes magnetic hybrid nanocomposites for a rapid and selective preconcentration and clean-up of mercury species in water samples.

Hybrid nanocomposites based on Fe3O4 magnetic nanoparticles (MNPs) coated with different types of carbon nanotubes (CNTs) have been studied for the first time as sorbent materials for magnetic solid phase extraction (MSPE) for mercury speciation analysis. Monomethylmercury (MMHg) was the target mercury species in water samples and the adsorption and desorption processes were optimized based on this species. Single-walled CNT-MNP showed higher adsorption capacity than double-walled or multi-walled CNTs. Then, the magnetic sorbent was retrieved with an external magnet and MMHg was selectively desorbed from it with dichloromethane (DCM) in two steps with vortex agitation. Inorganic mercury was removed during the desorption stage. The rapid adsorption and desorption equilibrium, the magnetic separation of the sorbent, and the simple and fast synthesis of CNT-MNPs without any additional modification of the CNTs simplified and shortened the extraction procedure. The extract was submitted to derivatization of the mercury species by ethylation (with an optional nitrogen stream evaporation of the organic phase) and injection into a gas chromatograph coupled to an atomic fluorescence detector (GC-pyro-AFS). The overall procedure provides the preconcentration of MMHg up to 150 times and the removal of inorganic mercury at the same time. The procedural limits of detection (LOD) and quantification (LOQ) were 5.4 and 17.9pgmL-1, respectively. Moreover, magnetic nanocomposites can be reused at least 7 times without losing their efficiency. The methodology was validated in tap, dam and river water samples to evaluate the performance under real conditions with recoveries from 79% to 97% of spiked MMHg.

Fast method for the simultaneous determination of monomethylmercury and inorganic mercury in rice and aquatic plants.

Recent investigations revealed that monomethylmercury (MMHg) can be absorbed and accumulated by plants, i.e. rice crops, thus becoming an important route of human exposure to MMHg through diet. The increasing concern about this fact makes that appropriate analytical methods for Hg speciation in these samples are urgently required. Therefore, the aim of this work has been the development of a fast and sensitive method which enables the simultaneous determination of MMHg and inorganic Hg in rice and aquatic plants. The proposed methodology is based on the extraction of Hg species by closed-vessel microwave heating, subsequent derivatization by ethylation and analysis by gas chromatography coupled to atomic fluorescence detection via pyrolysis (GC-pyro-AFS). A careful optimization of the extraction, using both acid (6N HNO3) and alkaline (tetramethylammonium hydroxide, TMAH) extractants, and derivatization conditions has been carried out. Spiked and unspiked aquatic plants (Typha domingensis) and CRMs certified for Total-Hg (BCR-60, BCR-482 and NCS ZC73027, corresponding to aquatic plant, lichen and rice, respectively) have been used. Under the final optimized conditions the simultaneous determination of MMHg and inorganic Hg can be carried out in less than 40min with no tedious clean-up steps. Quantitative recoveries (from 92% to 101%) were obtained in aquatic plants (Typha domingensis) and CRMs spiked with known concentrations of MMHg. For unspiked BCR-60 and BCR-482, no statistically significant differences (p=0.05) were found in Total-Hg concentrations between those obtained by the sum of species and the certified values for both acid and alkaline extraction. For the analysis of low Hg polluted samples, an additional preconcentration step by evaporation under nitrogen stream was required but adequate blanks were only obtained for acid extraction. Detection limits in the low ng/g range (0.7-1.0ng/g) were consequently achieved for both Hg species in the case of acid extraction and the analysis of NCS ZC73027 gave satisfactory results without statistically significant differences between the found and certified values (p = 0.05).

Large-scale projects in the amazon and human exposure to mercury: The case-study of the Tucuruí Dam.

The Tucuruí Dam is one of the largest dams ever built in the Amazon. The area is not highly influenced by gold mining as a source of mercury contamination. Still, we recently noted that one of the most consumed fishes (Cichla sp.) is possibly contaminated with methylmercury. Therefore, this work evaluated the mercury content in the human population living near the Tucuruí Dam. Strict exclusion/inclusion criteria were applied for the selection of participants avoiding those with altered hepatic and/or renal functions. Methylmercury and total mercury contents were analyzed in hair samples. The median level of total mercury in hair was above the safe limit (10µg/g) recommended by the World Health Organization, with values up to 75µg/g (about 90% as methylmercury). A large percentage of the participants (57% and 30%) showed high concentrations of total mercury (≥ 10µg/g and ≥ 20µg/g, respectively), with a median value of 12.0µg/g. These are among the highest concentrations ever detected in populations living near Amazonian dams. Interestingly, the concentrations are relatively higher than those currently shown for human populations highly influenced by gold mining areas. Although additional studies are needed to confirm the possible biomagnification and bioaccumulation of mercury by the dams in the Amazon, our data already support the importance of adequate impact studies and continuous monitoring. More than 400 hydropower dams are operational or under construction in the Amazon, and an additional 334 dams are presently planned/proposed. Continuous monitoring of the populations will assist in the development of prevention strategies and government actions to face the problem of the impacts caused by the dams.

Mapping the spatio-temporal risk of lead exposure in apex species for more effective mitigation.

Effective mitigation of the risks posed by environmental contaminants for ecosystem integrity and human health requires knowing their sources and spatio-temporal distribution. We analysed the exposure to lead (Pb) in griffon vulture Gyps fulvus-an apex species valuable as biomonitoring sentinel. We determined vultures' lead exposure and its main sources by combining isotope signatures and modelling analyses of 691 bird blood samples collected over 5 years. We made yearlong spatially explicit predictions of the species risk of lead exposure. Our results highlight elevated lead exposure of griffon vultures (i.e. 44.9% of the studied population, approximately 15% of the European, showed lead blood levels more than 200 ng ml(-1)) partly owing to environmental lead (e.g. geological sources). These exposures to environmental lead of geological sources increased in those vultures exposed to point sources (e.g. lead-based ammunition). These spatial models and pollutant risk maps are powerful tools that identify areas of wildlife exposure to potentially harmful sources of lead that could affect ecosystem and human health.

Sources and fate of mercury pollution in Almadén mining district (Spain): Evidences from mercury isotopic compositions in sediments and lichens.

Variations in mercury (Hg) isotopic compositions have been scarcely investigated until now in the Almadén mining district (Spain), which is one of the most impacted Hg areas worldwide. In this work, we explore and compare Hg isotopic signatures in sediments and lichens from Almadén mining district and its surroundings in order to identify and trace Hg aquatic and atmospheric contamination sources. No statistically significant mass independent fractionation was observed in sediments, while negative Δ(201)Hg values from -0.12 to -0.21‰ (2SD = 0.06‰) were found in lichens. A large range of δ(202)Hg values were reported in sediments, from -1.86 ± 0.21‰ in La Serena Reservoir sites far away from the pollution sources to δ(202)Hg values close to zero in sediments directly influenced by Almadén mining district, whereas lichens presented δ(202)Hg values from -1.95 to -0.40‰ (2SD = 0.15‰). A dilution or mixing trend in Hg isotope signatures versus the distance to the mine was found in sediments along the Valdeazogues River-La Serena Reservoir system and in lichens. This suggests that Hg isotope fingerprints in these samples are providing a direct assessment of Hg inputs and exposure from the mining district, and potential information on diffuse atmospheric contamination and/or geochemical alteration processes in less contaminated sites over the entire hydrosystem. This study confirms the applicability of Hg isotope signatures in lichens and sediments as an effective and complementary tool for tracing aquatic and atmospheric Hg contamination sources and a better constraint of the spatial and temporal fate of Hg released by recent or ancient mining activities.