Metal speciation

An analytical methodology to determine the molecular weight (MW) distribution patterns of several elements among different compounds present in commonly consumed edible mushrooms is presented in this work. A hyphenated technique based on size exclusion liquid chromatography (SEC) coupled on-line to UV and inductively coupled plasma mass spectrometry (ICP-MS) detection was used. The association of the elements to high and low MW fractions was confirmed with sequential detection by UV and ICP-MS. Separation of the fractions was performed by injecting a 100 µL sample volume to a Superdex 75 column. The effect of different mobile phases on the separation was evaluated. Additionally, three different extraction conditions including 0.05 mol L -1 NaOH, 0.05 mol L -1 HCl, and hot water at 60°C were applied to extract the elemental species from the mushroom samples. Significant differences were observed in the chromatograms depending on the extraction conditions utilized. Optimization of the experimental variables involved in the SEC-UV-ICP-MS coupling was carried out. The method was applied to investigate the fractionation patterns of Bi, Co, Cu, Fe, I, Mo, Ni, Se, and Zn in fungi porcini (Boletus edulis) mushroom. The results obtained in this work indicate an important association of most of the elements to high MW fractions.

Background, aim, and scope In water systems, water quality and geochemical properties of sediments determine the speciation of trace metals, metal transport, and sediment-water exchange, influencing metal availability and its potential effects on biota. Studies from temperate climates have shown that iron-ore mining and tailing wastewaters, besides being a source of trace metals, usually show high levels of dissolved ions and particulate suspended matter, thus having the potential of indirectly changing metal bioavailability. For the first time in the tropics, we identified the effects of iron-ore mining and processing on metal bioavailability in a coastal lagoon. With an extensive sampling scheme, we investigated the potential sources of metals; the links among metal levels in water, sediments, and invertebrates; and the contrasting effects on metal speciation and bioavailability. Methodology The metals Fe, Mn, Al, Cr, Zn, Cu, Ni, Pb, Cd, Hg, and As were measured in water, sediments (surface and profiles), and invertebrates from Mãe-Bá Lagoon and in the sites directly influenced by the mining operations (tailing dams and nearby rivers). In addition, samples from two other lagoons, considered pristine, were analyzed. The study area is located in the southeast of Brazil (Iron Quadrangle Region and a coastal area of Espírito Santo State). General water characteristics included pH, dissolved organic carbon, alkalinity, and anion composition. Water metal speciation was assessed by a speciation model (Chemical Equilibria in Aquatic Systems). Grain-size distribution, organic carbon, carbonate, and acid volatile sulfide (AVS) were determined in sediments. Statistical methods included comparison of means by Mann-Whitney test, ordination and correlation analyses, and analysis of regression for geochemical normalization of metals with grain size. Results and discussion The dissolved metal concentrations, the total metal levels in sediments, and the normalization based on the fine sediment fraction showed that the mining operations constitute potential sources of Fe, Mn, Cr, Cu, Ni, Pb, As, and Hg to Mãe-Bá Lagoon. However, trace metal availability was reduced because of increased pH, hardness, and sulfide content (356 μmol/g) in the sites influenced by the mining. The lagoon showed similar water chemistry as in the mining sites, with metal bioavailability further decreased by the presence of dissolved organic carbon and chloride. Although AVS levels in the lagoon were low (0.48-56 μmol/g), metal bioavailability was reduced because of the presence of organic matter. Metal levels in invertebrates confirmed the predicted low metal bioavailability in Mãe-Bá Lagoon. The lagoon was considered moderately contaminated only by Hg and As. Conclusions The iron-ore mining and processing studied here constitute potential sources of metal pollution into the tropical lagoon. Contrary to expectations, however, it also J Soils Sediments (contributes to reducing the overall metal bioavailability in the lagoon. Recommendations and perspectives These findings are believed to be useful for evaluating metal exposure in a more integrated way, identifying not only the sources of pollution but also how they can affect the components involved in metal speciation and bioavailability in water systems, leading to new insights.

Seasonal monitoring was carried out to investigate the influence of extracellular enzymatic activity (EEA) on metal speciation and organic matter cycling in the rhizosediment of Spartina maritima. Heavy metal speciation was achieved by the Tessier scheme, and showed a similar pattern of variation of the organicbound fraction, indicating a decomposition process in progress. Both humic acid and organic matter showed the same seasonal pattern. The basal respiration of the rhizosediments also presented a similar seasonal pattern, indicating a microbial degradation of organic matter. The high organic-bound fraction found in the summer gradually decreased towards the winter. This decrease was found to be related to the increase of activity of peroxidase, b-N-acetylglucosaminidase and protease. Also the activity of sulphatase was found to be related to the depletion on the exchangeable fraction, probably due to sulphide formation and consequent mobilization. The results show an interaction between several microbial activities, affecting metal speciation. Marine Ecology. ISSN 0173-9565 Marine Ecology 30 (Suppl. 1) (2009) 65-73 ª 2009 No claim to original government work Spartina maritima rhizosediment EEA and metal speciation Duarte, Almeida & Caç ador

After performing this experiment, the student shall be able to: •  Verify experimentally many of the transitions in metal Pourbaix diagrams. •  Understand the role of potential in metal speciation. •  Understand the role of pH in metal speciation.

The objective of this paper was to review the status of the Malaysian aquatic environment with respect to heavy metals. Much of the work on metals focused on measurements in biota and sediments, but very little research has been done on dissolved and particulate metals in water, metal speciation in sediments or on the bioavailability of metals. Cadmium (Cd), Copper (Cu), Lead (Pb) and Zinc (Zn) received much attention but due to technical difficulties, Arsenic, (As), Mercury (Hg), Tin (Sn) and other hydride elements have not been well studied.

Recent studies have demonstrated that the Guadiana Estuary contains metal concentrations in excess of background values. Therefore, this work aims to document the potential environmental hazards associated with the availability of these metals in this environment of high ecological value. Mineralogical analysis shows that the sediments are composed mainly of quartz, albite, and clay minerals (illite, smectite, kaolinite, and vermiculite) along with several small, reactive compounds (including soluble sulphated salts, Fe-Mn oxyhydroxides, organic matter, and pyrite) capable of retaining metals, which can be subsequently released, causing environmental degradation. BCR sequential extraction shows that As, Cd, Cu, Mn, Pb, and Zn present mobile fractions with respect to the total metal content (41, 100, 57, 53, 70, and 69%, respectively) in any of the described reactive phases (F1 + F2+ F3).Calculated environmental risk indices demonstrate moderate to considerable ecological risk for almost the entire estuary, associated mainly with acid mine drainage from the nearby Iberian Pyrite Belt. In addition, the indices highlight several zones of extremely high risk, which are related to industrial and urban dumps in the vicinity of the estuary and to heavy traffic on the international bridge.

Biosorption data of lead, copper, zinc and cadmium onto Sphaerotilus natans at different equilibrium pH (3-5 units) were here reported and analysed. Experimental results outlined the positive effect of pH increase on pollutant uptake and also the biomass affinity series (Pb>Cu>Zn>Cd) reflecting the hydrolytic properties of metals. An original empirical model was proposed to represent the effect of pH on heavy metal biosorption inserting q max vs. pH empirical functions into the classical Langmuir isotherm. r

A chelating resin containing iminodiacetic groups, Chelex 100 which is often employed for sorbing metal ions from complex matrices, was used for separating and preconcentrating heavy metal ions from seawater samples. The resin titration method was applied for determining the total concentration. The experimental procedure consists of measuring the concentration of the metal sorbed on Chelex 100 from solutions with different ratios volume of aqueous phase/amount of resin. The total metal ion concentration is obtained from the experimental data by a model for the sorption on the resin, the parameters of which are obtained by a non-linear regression procedure. In this way not only the total metal ion concentration can be determined, but also the side reaction coefficient α M(I) , if the sorption equilibria of the considered metal ion on the resin are exactly known, as is the case of Chelex 100 considered here. The resin titration with Chelex 100 was utilized for determining the total metal ion concentration in certified seawater samples and real samples from the Ligurian Sea. It allowed the simultaneous determination of the total concentration of manganese(II), cadmium(II), nickel(II) and copper(II), in acceptable agreement with the certified values. Moreover, the side reaction coefficients of some complex species could be evaluated. They were very high, and strongly dependent on the acidity of the sample solution. For instance, values varying from 10 9 to 10 11 were obtained for copper (II) in the different samples considered, at pH around 7. Chelex 100 is such a strong sorbing substance that it is able to compete with the stable complexes for the combination with the heavy metal ions considered here. (M. Pesavento) 0003-2670/99/$ -see front matter ©1999 Elsevier Science B.V. All rights reserved. PII: S 0 0 0 3 -2 6 7 0 ( 9 9 ) 0 0 5 0 6 -1

Cadmium, chromium, copper, lead, manganese, nickel, vanadium and zinc have been determined in samples of made-up ground from layers of a trial pit excavated on a recently derelict, industrially contaminated site. The pseudototal metal content of the layers was determined following a microwave-assisted digestion with aqua regia. Operational speciation was performed using the BCR three-step sequential extraction procedure. Analyses were carried out by¯ame or electrothermal atomic absorption spectrometry (FAAS or ETAAS). A suppressive interference ($30%) was observed in the determination of cadmium in aqua regia digests by ETAAS, but not in 0.11 mol l À1 acetic acid, 0.1 mol l À1 hydroxylammonium chloride or 1.0 mol l À1 ammonium acetate extracts. Agreement between duplicate samples was acceptable (i.e. within 10%) for most elements in most layers, but some large discrepancies were apparent, especially for lead. The amount of metal extracted in the sequential procedure (i.e. Step 1Step 2Step 3residual) did not generally agree well with pseudototal digestion. Various layers of the trail pit contained signi®cant levels of contaminant metals, but these were not always in easily mobilized forms. For example, less than 0.2% of the lead ($4000 mg g À1 ) at 65±85 cm depth was present as exchangeable or acid-soluble species. The study illustrates the importance of considering metal speciation when assessing the mobility of potentially toxic elements in industrially-contaminated land. # 1998 Elsevier Science B.V.

Despite the growing evidence that the vapor phase, formed through magma degassing and ore fluid boiling, can selectively concentrate and transport metals, the effects of major volatile components like sulfur, chlorine or carbon dioxide on the metal vapor-liquid fractionation and vapor-phase transport under magmatic-hydrothermal conditions remain poorly known. We performed systematic experiments to investigate the effect of sulfur ligands on metal vapor-liquid partitioning in model H 2 O-S-NaCl-KCl-NaOH systems at temperatures from 350 to 500°C. Results show that at acidic-to-neutral conditions, vapor-liquid equilibrium distribution coefficients, K m = m vapor / m liquid , where m is the mass concentration of the metal in corresponding phase, of metalloids (As, Sb) and base metals (Zn, Fe, Pb, Ag) are in the range 0.1-1.0 and 0.001-0.1, respectively, and are not significantly affected by the presence of geologically common sulfur concentrations, up to 1-3 wt.% S. In contrast, the partitioning of Cu, Au, and Pt into the vapor increases by a factor of 100 in comparison to the S-free water-salt system, yielding K m values of 0.5-1.0, 1-10, and 10-20, respectively, due to formation of volatile neutral complexes with H 2 S and, possibly, SO 2 . In neutral-to-basic systems, Zn, Pb, Fe and Ag show 10-100-fold increase of their partition coefficients, whereas Cu, Au and Pt exhibit K m values of up to several orders of magnitude lower, compared to acidic conditions at similar temperature, pressure and sulfur contents. These vapor-liquid distribution patterns result from combined effects of i) formation of volatile species with reduced sulfur ligands in the vapor phase, ii) changes in the metal speciation in the coexisting liquid phase as a function of pH, and iii) solute-solvent interactions in both phases. Our data explain the vapor-liquid fractionation trends for many metals as inferred in coexisting brine and vapor inclusions from magmatic-hydrothermal deposits, and provide a first experimental evidence for the dramatic increase of the mobility of Cu, Au and Pt in sulfur-enriched acidic magmatic-hydrothermal vapors, consistent with geological models of Au± Cu ores formation and distribution in porphyry-epithermal settings.

Different experimental approaches have been suggested in the last few decades to determine metal species in complex matrices of unknown composition as environmental waters. The methods are mainly focused on the determination of single species or groups of species.

The application of sequential extraction procedures to determine metal speciation in sediments is fraught with uncertainty regarding what is actually dissolving or re-precipitating at each stage. In order to choose an appropriate scheme for the investigation of contaminated anaerobic mud two different sequential extraction procedures (Kersten and Förstner, 1986; Quevauviller, 1998) were investigated using a Cryogenic SEM (CryoSEM) technique coupled

Bioavailability considerations are one of the tools for a proper assignment of sites potentially and actually at risk as it allows assessing both the extent (hazard) and probability (risk) of adverse effects. In this paper, bioavailability considerations are linked to physicochemical methods available for assessing metal fractions in soils. The focus of the overview is on empirical methods for extraction of metals from soils as a surrogate for the metal-, species-and soil-type-dependent bioavailable and bioaccessible metal pools. This cumulates in a generalized flow chart for monitoring of metals in soils. In support of the general monitoring strategy, examples are given of successful applications of analytical methods for predicting metal uptake by plants and animals, for assessing the origin of metals in soils, as well as the leaching potential of soils and the extent of soil contamination. It is concluded that a large arrays of chemical assessment methodologies for metal speciation in solid and liquid soil phases are available. As most assessment methodologies are operationally defined instead of being functionally defined, examples of mechanistically based monitoring approaches of bioavailability are still scarce. The value of the methods for measuring bioavailability can be significantly improved when the species, metal, and soil specific aspects of bioavailability are more accurately taken into account in the design of chemical simulation methodologies. r

SRM 1974a, Organics in Mussel Tissue (Mytilus edulis); SRM 2974, Organics in Mussel Tissue (freeze-dried); and SRM 2976, Mussel Tissue (trace elements and methylmercury) have been recently certified for methylmercury and total mercury content. Three independent analytical procedures were used to determine the certified methylmercury concentrations. Four independent procedures combined with data from two intercomparison exercises were used to determine the certified total mercury concentrations. These materials are the first certified metal speciation environmental SRMs issued by the National Institute of Standards and Technology.

During the time-course of ecotoxicity tests with algae and chronic (reproductive) toxicity tests with daphnids, in which algae are present as a food source, pH can dramatically increase due to photosynthetic activity. As pH changes can significantly affect metal speciation and thus its bioavailability, it may be necessary to buffer the pH of the exposure medium. One class of buffers (Good's N-subtituted aminosulfonic acids) are increasingly being used in biological and chemical applications, including ecotoxicity testing. However, the potential effect of these buffers on metal toxicity has, so far, scarcely been examined. In this study we investigated if MOPS (3-N morpholino propane sulfonic acid) affected the toxicity of copper and zinc to two standard test organisms: the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata. First, we demonstrate that up to a concentration of 750 mg l )1 (which proved to be sufficient for pH buffering) MOPS did not affect 21-day net reproduction of D. magna or the 72-h population growth of P. subcapitata. Second, we conducted bioassays in copper and zinc spiked standard media for the pH range 6 -8. For D. magna the possible effect of 750 mg l )1 MOPS on acute copper and zinc toxicity was investigated by performing parallel 48-h toxicity tests in NaHCO 3 and MOPS buffered test media. Seventy-two hour growth inhibition assays with P. subcapitata were performed in parallel in MOPS and NaHCO 3 buffered test media and in test media with daily manual pH adjustment with HCl. For daphnids no significant differences in copper and zinc toxicity were observed between MOPS or NaHCO 3 buffered test media. For algae no significant differences in metal toxicity were observed between MOPS and HCl buffered media, but in test media buffered with NaHCO 3 an increased copper and zinc toxicity was observed as a consequence of pH increases during the test. Clearly, the results of this study demonstrate the importance of pH buffering in metal toxicity testing and the suitability of the MOPS buffer for that purpose.

An industrial site, polluted with As and heavy metals, was investigated by combining chemical (sequential extractions and pH leaching tests), physical and mineralogical characterization of soil samples and slag fragments, stat and by the analysis of soil porewater aimed at assessing the distribution, speciation and mobility of heavy metals and As. On the site itself, arsenic concentrations up to 3.6% in surficial soil samples and up to 22% in slag fragments were found, together with elevated concentrations (percentage level) of Cu, Co, Ni, Zn and Pb. High concentrations of arsenic (up to 38 000 mgyl) and heavy metals (up to 1700 mgyl Cu and 4700 mgyl Zn) were also found in the in situ sampled soil porewater, highlighting the considerable availability of As, Zn and Cu for uptake by plants and leaching to the ground water. Sequential extractions also indicated a high availability of arsenic and copper in most samples and slag fragments of the industrial site, although poorly reactive phases were encountered as well. pH stat leaching tests confirmed that the present leaching of contaminants is alarming. Moreover, soil acidification will enhance the leaching of contaminants, emphasizing that remediation of the industrial site is urgent. Small scale variability of total metal concentrations and metal speciation, both in the horizontal and vertical direction, and the occurrence of a camouflage layer underline the importance of elaborate sampling for pollution assessment on an industrial site. ᮊ

A systematic investigation evaluated the concentrations of a selected number of trace elements (Cd, Co, Cu, Li, Ni, Pb, V and Zn) in carbonates of the benthic foraminifera Ammonia tepida collected from surface sediments of the highly polluted harbour of Naples. Application of cleaning procedures, combined with Scanning Electron Microscopy investigation (SEM) of the analysed shells allowed reliable quantification of the elements in the carbonate lattice. Adoption of biogenic carbonate/seawater distribution coefficients reported in the literature provided the ranges of variability of total dissolved trace elements in the studied marine environment. Very high concentrations of Zn, Cd, and Cu calculated in seawater (from 100 to 10,000 times higher than those reported for uncontaminated Mediterranean seawaters) testify to intense effects of anthropogenic impact on the harbour mainly related to the industrial and commercial activities carried out in the neighbouring area. The ensemble of the obtained results emphasizes the high potential of measurements of trace elements in the biogenic carbonates of benthic foraminifera as tracers of anthopogenic pollution of seawater and reliable proxies of potentially bioavailable forms (as free ions and/or more labile organic complexes) of seawater dissolved metals.

An industrial site, polluted with As and heavy metals, was investigated by combining chemical (sequential extractions and pH leaching tests), physical and mineralogical characterization of soil samples and slag fragments, stat and by the analysis of soil porewater aimed at assessing the distribution, speciation and mobility of heavy metals and As. On the site itself, arsenic concentrations up to 3.6% in surficial soil samples and up to 22% in slag fragments were found, together with elevated concentrations (percentage level) of Cu, Co, Ni, Zn and Pb. High concentrations of arsenic (up to 38 000 mgyl) and heavy metals (up to 1700 mgyl Cu and 4700 mgyl Zn) were also found in the in situ sampled soil porewater, highlighting the considerable availability of As, Zn and Cu for uptake by plants and leaching to the ground water. Sequential extractions also indicated a high availability of arsenic and copper in most samples and slag fragments of the industrial site, although poorly reactive phases were encountered as well. pH stat leaching tests confirmed that the present leaching of contaminants is alarming. Moreover, soil acidification will enhance the leaching of contaminants, emphasizing that remediation of the industrial site is urgent. Small scale variability of total metal concentrations and metal speciation, both in the horizontal and vertical direction, and the occurrence of a camouflage layer underline the importance of elaborate sampling for pollution assessment on an industrial site. ᮊ

The bioavailability and therefore toxicity of a metal depends on the chemical species present in a particular environment. We evaluated the effect of a series of factors that could potentially modify metal speciation on the toxicity of Hg, Cu, Zn, and Cd toward a recombinant strain of the freshwater cyanobacterium Anabaena sp. PCC 7120 with cloned lux operon of luminescent terrestrial bacterium Photorhabdus luminescens. The strain, denoted as Anabaena CPB4337, showed a high constitutive luminescence with no need to add exogenous aldehyde. The tested factors were pH, EDTA (as organic ligand), and anions PO 4 3-, CO 3 2-, and Cl -. Chemical modeling and correlation analyses were used to predict metal speciation and link it with toxicity. In general, metal toxicity significantly correlated to the predicted metal free-ion concentration, although Zn-EDTA complexes and certain Hg chloro-complexes could also exhibit some toxicity to cyanobacteria. An interesting feature of metal toxicity to strain Anabaena CPB4337 was that low amounts of PO 4 3and CO 3 2increased metal toxicity; this effect could not be related to significant changes in metal speciation and could be attributed to a modulating effect of these anions on metal/uptake toxicity. The combination of toxicity studies that take into account a range of factors that might modulate metal toxicity with chemical modeling to predict changes in metal speciation might be useful for interpreting complex toxicity data. Finally, this cyanobacterial bioreporter, due to its ecological relevance as a primary producer, could be used as a tool for toxicity assessment in freshwater environments.

The present work deals with the application of biotechnology for the mobilization of metals from different solid wastes: end of life industrial catalysts, heavy metal contaminated marine sediments and fluorescent powders coming from a cathode ray tube glass recycling process. Performed experiments were aimed at assessing the performance of acidophilic chemoautotrophic Fe/S-oxidizing bacteria for such different solid matrices, also focusing on the effect of solid concentration and of different substrata. The achieved results have evidenced that metal solubilization seems to be strongly influenced by the metal speciation and partitioning in the solid matrix. No biological effect was observed for Ni, Zn, As, Cr mobilization from marine sediments (34%, 44%, 15%, 10% yields, respectively) due to metal partitioning. On the other hand, for spent refinery catalysts (Ni, V, Mo extractions of 83%, 90% and 40%, respectively) and fluorescent powders (Zn and Y extraction of 55% and 70%, respectively), the improvement in metal extraction observed in the presence of a microbial activity confirms the key role of Fe/S oxidizing bacteria and ferrous iron. A negative effect of solid concentration was in general observed on bioleaching performances, due to the toxicity of dissolved metals and/or to the solid organic component.

The use of organic waste and compost as a source of organic matter and nutrients is a common practice to improve soil physico-chemical properties, meanwhile reducing the need for inorganic fertilisers. Official guidelines to assess sewage sludge and compost quality are mostly based on total metal content of these residues. Measurement of the total concentration of metals may be useful as a general index of contamination, but provides inadequate or little information about their bioavailability, mobility or toxicity when the organic residue is applied to the soil. However, ecotoxicity tests provide an integrated measure of bioavailability and detrimental effects of contaminants in the ecosystem. In the present study, three different types of biodegradable organic residues (BORs) have been considered: sewage sludge from municipal wastewater treatment (SS), compost from the organic fraction of unsorted municipal solid waste (MSWC), and garden waste compost (GWC). The BORs were subjected to chemical characterisation and total metal quantification (Cd, Cr, Cu, Ni, Pb and Zn), in order to verify their suitability for land application. Water leachability was determined through the DIN 38414-S4 method, while the modified BCR sequential extraction procedure was used for metal speciation. Ecotoxicity of the BORs was studied by direct and indirect bioassays. Direct toxicity bioassays were: plant growth tests with cress (Lepidium sativum L.) and barley (Hordeum vulgare L.), and earthworm (Eisenia fetida) mortality. On the other hand, indirect exposure bioassays, with leachate from the residues, took into account: luminescent bacteria (Vibrio fischeri), seed germination (L. sativum and H. vulgare) and Daphnia magna immobilization. As far as total metal concentration is concerned, with particular reference to Zn, SS resulted neither suitable for the use in agriculture nor compatible to be disposed of as an inert material into landfill, according to the Directive 1999/31/EC. Zinc in SS was mainly present in exchangeable form (28.5%), appearing as highly bioavailable. As a consequence, SS exhibited either high ecotoxicity effects with the indirect exposure bioassays or significant mortality with the earthworm bioassay. Total content of metals in MSWC allowed its classification as "stabilised biowaste", according to 2nd draft [DG Env.A.2. Working document of Biological treatment of biowaste -2nd draft. Directorate-General Environment, Brussels, 12th February; 2001. accessed in:http://europa.eu.int/comm/environment/waste/facts_en.htm, at 10/09/2002] while leachate, on the basis of the concentration of these contaminants, could be classified as "inert waste". This residue showed significant ecotoxicity effects with direct exposure bioassays as well as with the luminescent bacteria bioassay. However, it resulted less toxic than SS. Finally, GWC could be classified as a Class 2 compost, with no detectable toxic effects on the organisms used in the bioassays, except for the luminescent bacteria. In this case, an EC 50 of 73.0% was observed. Considering the results, the use of a battery of toxicity test in conjunction with chemical analysis should be suggested, in order to correctly assess possible environmental risks deriving from disposal or land application of biodegradable organic residues.

The objective of this paper was to review the status of the Malaysian aquatic environment with respect to heavy metals. Much of the work on metals focused on measurements in biota and sediments, but very little research has been done on dissolved and particulate metals in water, metal speciation in sediments or on the bioavailability of metals. Cadmium (Cd), Copper (Cu), Lead (Pb) and Zinc (Zn) received much attention but due to technical difficulties, Arsenic, (As), Mercury (Hg), Tin (Sn) and other hydride elements have not been well studied. The sources of metal pollution come from manufacturing, agriculture, sewage and motor vehicle emissions. The few reliable studies on metals in water indicate that some rivers were contaminated with Pb, Cu and Zn. Measurements in fish, bivalves and shrimp indicate no contamination of these resources by metals. However contamination by Cd and Pb was noted in some littoral shore molluscs such as Thais sp. as well as rock oysters (Saccostrea sp.) which may be due to non-anthropogenic sources. Sediments of the Juru and Langat rivers are contaminated by Pb and Zn, while the Langat River was heavily contaminated by Cd. The concentrations of Zn and Pb in coastal sediments off Juru in Penang, and in the Johor Strait were two and three times higher than global shale values. The source of Pb pollution was thought to be derived from the use of leaded petrol. Metal levels in sediments of the Malacca Strait and the South China Sea were generally similar to global shale values.

Varying the counter-ion is a highly supportive practice in tackling the problem of poor water-solubility of metal complexes of pharmaceutical importance. As a matter of fact, the relevant structural modification may alter the metabolic pathways and possibly the mode of action of a drug. To prove that this does not take place for one of the lead anticancer metal-based developmental compounds, indazolium trans-[RuCl 4 (1H-indazole) 2 ] (KP1019), its reactivity toward human serum proteins was assessed under simulated physiological conditions and compared to that of a much more soluble analogue, sodium trans-[RuCl 4 (1H-indazole) 2 ] (KP1339). For such kinetic assaying, capillary electrophoresis (CE) interfaced online with inductively coupled plasma mass spectrometry (ICP-MS) to specifically monitor changes in the metal speciation following the formation of ruthenium-protein adducts was applied. The rate constants of interaction with albumin and transferrin were determined at pharmacologically fitting drug-to-protein ratios as on average 0.0319 ± 0.0021 min −1 and 0.0931 ± 0.0019 min −1 (KP1019) and 0.0316 ± 0.0018 min −1 and 0.0935 ± 0.0053 min −1 (KP1339), respectively. The results of this brief study showed that changing from organic to inorganic counter-ion at the stage of formulation could commonly be recommended for improving ruthenium-based drug solubility and bioavailability. (A.R. Timerbaev).

A knowledge of the total content of trace metals is not enough to fully assess the environmental impact of polluted soils. For this reason, the determination of metal species in solution is important to evaluate their behaviour in the environment and their mobilization capacity. Sequential extraction procedure was used to speciate five heavy metals (Cd, Pb, Cu, Ni and Zn) from four contaminated soils of Southern Nigeria into six operationally defined geochemical species: water soluble, enchangeable, carbonates, Fe-Mn oxide, organic and residual. Metal recoveries were within +/- 10% of the independently determined total Cd, Pb, Cu, Ni and Zn concentrations. The highest amount of Cd (avg. 30%) in the nonresidual fractions was found in the exchangeable fraction, while Cu and Zn were significantly associated with the organic fraction. The carbonate fraction contained on average 14, 18.6, 12.6, 13 and 11% and the residual fraction contained on average 47, 18, 33, 50 and 25% of Cd, Pb, Cu...

The elemental composition of seasoning products, like spices garlic and onion, has been reviewed paying special attention to methods proposed in the literature to determine specific elements or as much as possible components of these products which are widespread used in food preservation or food flavouring and can contribute to the intake of both, essential and toxic elements. Sample treatment and metal speciation aspects have been critically revised concerning onion, garlic and pepper (the most commonly studied products).

This work aimed to evaluate, in vitro, the capability of roots of salt marsh plants to release strong Cu-complexing ligands and to ascertain whether Cu contamination would stimulate ligands' exudation or not. The sea rush Juncus maritimus and the sea-club rush Scirpus maritimus, both from the lower Douro river estuary (NW Portugal), were used. Plants were collected seasonally, four times a year in 2004, during low tide. After sampling, plant roots were washed for removal of adherent particles and immersed for 2 h in a solution that matched salinity (3) and pH (7.5) of the pore water from the same location and spiked with Cu 2þ in the range 0e1600 nM to obtain plant exudates. In the final solutions as well as in sediment pore water total dissolved Zn and Cu, Cu-complexing ligand concentrations and the respective conditional stability constants (K CuL 0 ) values were determined by voltammetry. This study demonstrated that plants are able to release, in a short period of time, relatively high amounts of strong Cu-complexing ligands (56e265 nmol g root À1 ), which differed among plants and sampling site but were independent of the season. Cu contamination did not stimulate exudation of Cu-complexing ligands. On the other hand, in media contaminated with Cu both plants accumulated relatively high amounts (29e83%) of the initially dissolved Cu, indicating that they have alternative internal mechanisms for Cu detoxification. Cu exchange between roots and medium (either accumulation in contaminated medium or release in the absence of Cu) was more intense for S. maritimus than for J. maritimus. It was observed that exudate solutions obtained in the absence of added Cu and sediment pore water (the densities of roots observed inside the salt marsh where comparable to those used in the in vitro experiments), displayed similarities in terms of total dissolved metals, Cu-complexing ligands concentrations, values of K CuL 0 (12 < log K CuL 0 < 14), as well as patterns of variation among seasons (only observed for Zn). These results are novel and point out that salt marsh plants may be the source at least partially of the strong organic ligands found in the sediment pore water in shallow marginal areas. The capability of salt marsh plants to release strong organic ligands into the environment, conjugated with their known capacity to oxidize anaerobic sediment around roots, indicate that these plants can play a role in controlling metal speciation in the water/sediment interface.

Water samples of contrasting origin, including natural seawater, two sediment elutriates and sewageinfluenced seawater, were collected and obtained to examine the effect of the dissolved organic matter (DOM) present on metal bioavailability. The carbon content (DOC) and the optical properties (absorbance and fluorescence) of the coloured DOM fraction (CDOM) of these materials were determined. Cu and Pb complexation properties were measured by anodic stripping voltammetry (ASV) and the effect of DOM on Cu and Pb bioavailability was studied by means of the Paracentrotus lividus embryo-larval bioassay. Sediment elutriates and sewage-influenced water (1) were enriched 1.4-1.7 times in DOC; (2) absorbed and reemitted more light; and (3) presented higher Cu complexation capacities (L Cu ) than the natural seawater used for their preparation. L Cu varied from 0.08 M in natural seawater to 0.3 and 0.5 M in sediment elutriates and sewage-influenced water, respectively. Differences in DOC, CDOM and Cu complexation capacities were reflected in Cu toxicity. DOM enriched samples presented a Cu EC 50 of 0.64 M, significantly higher than the Cu EC 50 of natural and artificial seawater, which was 0.38 M. The protecting effect of DOM on Cu toxicity greatly disappeared when the samples were irradiated with high intensity UV-light. Cu toxicity could be successfully predicted considering ASV-labile Cu concentrations in the samples. Pb complexation by DOM was only detected in the DOM-enriched samples and caused little effect on Pb EC 50 . This effect was contrary for both elutriates: one elutriate reduced Pb toxicity in comparison with the control artificial seawater, while the other increased it. UV irradiation of the samples caused a marked increase in Pb toxicity, which correlated with the remaining DOC concentration. DOM parameters were related to Cu speciation and toxicity: good correlations were found between DOC and Cu EC 50 , while L Cu correlated better with the fluorescence of marine humic substances. The present results stress the importance of characterizing not only the amount but also the quality of seawater DOM to better predict ecological effects from total metal concentration data.

Trace metals pollution of surface waters and their nearby soils in the metallurgically polluted Kardjali region, Bulgaria, were studied. Thermodynamic model-ing including the dissolved organic carbon (DOC) was carried out for evaluating the distribution of metal species in waters and soil solutions. Zn was found to be the most widespread pollutant in the water samples, followed by Cu, Mn, and Cd. Geoaccumulation indices of trace metals for the tested soils were calculated, indicating that regarding Al, Fe, Co, Ni, and Cu all soils are Buncontaminated^. The most significant soil pollutant was found to be Cd, with all soils being either Bextremely contaminated^by this metal or close to the limit, followed by Pb and Zn. The dynamics of trace metal chemical species distribution in surface waters and in the water-soluble soil fractions, as a result of possible spontaneous precipitations, was calculated by applying different thermodynamic models. Regarding Mn, Co, Ni, and Cd in waters and aqueous soil extracts and Zn in aqueous soil extracts, their free ion species prevailed, being more labile and hence toxic for the ecosystem. In the case of Al, Fe, Cu, Cd, and Pb in the waters and aqueous soil extracts and of Zn in waters, stable organic complexes with bidentate bonds, Me(OH) 4 − or Me(OH) 0 2 prevailed.

Trace metal speciation in seawater from the Bulgarian Black Sea coast was studied in situ by hollow fiber permeation liquid membrane (HF-PLM) and by diffusion gradients in thin-film gels (DGT). The concentrations of Cd, Cu, Ni, and Pb determined by HF-PLM were lower than those measured by DGT, in agreement with their analytical windows, e.g., free metal ions provided by the HF-PLM and dynamic (mobile and labile) species by the DGT. The obtained suite of data was further used to evaluate the bioavailability of these metals to the microorganisms, which was then compared with experimental results of metal uptake to green microalga Chlorella salina. Uptake fluxes of the Cd, Cu, Ni, and Pb to C. salina, were predicted from the measured HF-PLM concentrations and laboratory experimentation in artificial seawater, in agreement with theoretical considerations. The HF-PLM and DGT appear to be promising analytical techniques for speciation and bioavailability studies in complex environmental media and allow improved understanding of the role of different chemical species in metal bioavailability (and impact) in seawaters.

This study characterises the heavy-metal content in leachates collected from eight landfills in France. In order to identify heavy metal occurrence in the different size fractions of leachates, a cascade filtration protocol was applied directly in the field, under a nitrogen gas atmosphere to avoid metal oxidation. The results of analyses performed on the leachates suggest that most of the metals are concentrated in the < 30 kD fraction, while lead, copper and cadmium show an association with larger particles. Initial speciation calculations, without considering metal association with organic matter, suggest that leachate concentrations in lead, copper, nickel and zinc are super-saturated with respect to sulphur phases. Speciation calculations that account for metal complexation with organic matter, considered as fulvic acids based on C1(s) NEXAFS spectroscopy, show that this mechanism is not sufficient to explain such deviation from equilibrium conditions. It is therefore hypothesized that the deviation results also from the influence of biological activity on the kinetics of mineral phase precipitation and dissolution, thus providing a dynamic system. The results of chemical analyses of sampled fluids are compared with speciation calculations and some implications for the assessment of metal mobility and natural attenuation in a context of landfill risk assessment are discussed.

The elemental composition of seasoning products, like spices garlic and onion, has been reviewed paying special attention to methods proposed in the literature to determine specific elements or as much as possible components of these products which are widespread used in food preservation or food flavouring and can contribute to the intake of both, essential and toxic elements. Sample treatment and metal speciation aspects have been critically revised concerning onion, garlic and pepper (the most commonly studied products).

Biotic ligand models have been developed for various metals (e.g. Cu, Ag, Zn) and different aquatic species. These models incorporate the effect of physico-chemical water characteristics (major cations, pH, dissolved organic carbon) on the bioavailability and toxicity of the metal. In this study, the individual effects of calcium, magnesium, potassium, sodium and pH on zinc toxicity to the green alga Pseudokirchneriella subcapitata (formerly and better known as Selenastrum capricornutum and Raphidocelis subcapitata) were investigated. Stability constants for binding to algal cells (K ) were derived for those cations affecting zinc toxicity, using the mathematical approach proposed by De BL Schamphelaere and Janssen wEnviron. Sci. Technol. 63, (2002) 48-54x. Potassium proved to be the only cation tested that did not alter zinc toxicity to algae significantly. Log (K ) values for Ca, Mg and Na, derived at pH 7.5, were 3.2, BL 3.9 and 2.8, respectively. Toxicity tests performed at different pH values (5.5-8.0) indicated that competition between H and Zn reduces zinc toxicity. However, the observed relationship between (H ) and the 72h-EbC wexpressed as q 2q q 50 mM (Zn )x is not linear and suggests that pH affects the physiology of the biotic ligand. Although, in general, our 2q findings seem to suggest that zinc toxicity to algae can be modelled as a function of key water characteristics, the results also demonstrate that the part of the conventional BLM-hypothesis-i.e. that the binding characteristics of the biotic ligand are independent of the test medium characteristics-is not valid for algae. The observed pH-dependent change of stability constants should therefore be further investigated and incorporated in future BL-modelling efforts with algae. ᮊ

An approach is described for viewing the interrelationship between different variables and also tracing the sources of pollution of groundwater of north Chennai (India). The data set of 43 variables which include major ions, minor ions and trace metal speciation (Cu, Pb, Cd and Zn) collected during the pre-monsoon and post-monsoon seasons of the year 2000-2001, was subjected to R-mode factor analysis to comprehend the distribution pattern of the said variables. It was found that first factor measures salinity and hardness which explained 19.12% of the total variance (comprised of variables EC, TDS, Na + , K + , Ca 2+ , Mg 2+ , total hardness, Cl − and SO 2À 4) during pre-monsoon, while it was 25.08% during postmonsoon. The second and third factors were attributed to speciation of zinc and copper ions during both premonsoon and post-monsoon. Although there were two more factors, loaded with speciation parameters of lead and cadmium, the variance of them were less than 10%. From this study it is seen that sea water intrusion, municipal solid waste disposal are the identified sources of component of pollution. The importance of metal ions is taking a secondary role and the anthropogenic origin-industrial activity, is the reason in the evaluation of pollution status as they come in the second, third, fourth and fifth factors. As the trace metal speciation was grouped in separate factors, linear regression model (LRM) with correlation analysis was applied to check its validity for prediction of speciation and to apply LRM for rapid monitoring of water pollution.

The United States Environmental Protection Agency (USEPA) has established nationally applicable water quality criteria (WQC) for metals that are designed to be protective of aquatic life. However, in some instances these criteria may be overprotective as a result of natural, site-speci®c dierences in water quality characteristics. These dierences aect metal speciation and bioavailability, fundamental considerations in assessing toxicity. Laboratory studies completed during recent years have advanced the current understanding of metal chemistry in aquatic systems, including the formation of organic and inorganic metal complexes and sorption to particulate organic matter. Parallel investigations have led to an improved understanding of the physiological basis of why metals are toxic to aquatic organisms. These studies have, in combination, led to an improved understanding of how site-speci®c water chemistry aects bioavailability, and how metals exert toxicity at the organism site of action, at the biotic ligand in the context of the model to be described. The biotic ligand model provides a quantitative framework for assessing metal toxicity over a range of hardness, pH and dissolved organic carbon (DOC) levels. The chemical equilibrium sub-model incorporates metal-biotic ligand interactions to compute metal accumulation at the site of action (e.g., the gill of a ®sh) as a function of water quality. The toxicity sub-model uses this computed accumulation level as the basis for successfully predicting observed variations in toxicity associated with changes in water quality. The results highlight the potential utility of this approach to provide an alternative means of developing site-speci®c permit limits and WQC. The ability of the model framework to quantitatively assess the eects of hardness, pH and DOC on toxicity, in comparison to current WQC for metals that typically vary with hardness alone, is a signi®cant advance in understanding how site-speci®c conditions aect the toxicity of metals. 7

Combining in situ diffusion and column ion-exchange equilibration, we measured free metal ion concentrations (Cd, Cu and Zn) in water samples collected from the epilimnion of 14 lakes in the Rouyn-Noranda area (600 km north-west of Montreal, QC, Canada). Lakes were selected to represent a wide range of physico-chemical characteristics (hardness, pH, dissolved organic matter-DOM, degree of metal contamination), to determine the influence of these parameters on metal speciation. Total dissolved metal concentrations, as determined within the diffusion cells, varied over one to two orders of magnitude: [Cd] 0.19-2.9 nM; [Cu] 36-190 nM; [Zn] 7-2,800 nM. The proportion of total dissolved metal present as free Cd 2? and Zn 2? was relatively constant for the 14 selected lakes, despite the wide pH (4.5-8) and DOM (3-23 mg C/L) ranges, probably reflecting the inverse relationship observed between pH and DOM; this proportion did, however, vary with DOM and pH for Cu. Our experimental free metal ion concentrations were compared with those calculated with the thermodynamic models WHAM (Windermere Humic Aqueous Model VI) and ECOSAT 4.7 (incorporating the NICA-Donnan model). Measured and calculated values were in reasonable agreement for both Cd and Zn although measured values were generally slightly higher, i.e. less than one order of magnitude. For several lakes, measured free Cu concentrations were, however, much higher than the calculated values, suggesting that these models overestimate Cu complexation. The gap between measured and calculated free metal ion concentration becomes more important as the total metal concentration decreases and as pH increases.

The term bioavailability has many different meanings across various disciplines of toxicology and pharmacology. Often bioavailability is concerned with human health aspects such in the case of lead (Pb) ingestion by children. However, some of the most contaminated sites are found in nonpublic access facilities (Department of Defense or Energy) or in remote regions as a result of mining or industrial practices in which ecoreceptors such as plants, animals, and soil organisms are the primary concerns as well as the potential for food-chain transfer. In all cases, the endpoint requires movement of the element across a biological barrier. The still utilized approach to base risk assessment on total metal content in soils is an outdated endeavor and has never been proved to be scientifically sound. Yet to reverse this trend, much work is required to establish baseline bioavailability measurements and to develop complementary methods that are capable of predicting bioavailability across a whole range of impacted media in a cost-efficient manner. Thus, regulators have recognized site-specific human health risk assessments play a key role in decision-making processes at contaminated sites.

Determination of only total element in sediments does not give an accurate estimate of the likely environmental impacts. Speciation study of metals in sediment provides information on the potential availability of metals (toxic) to biota under various environmental conditions. In water, the toxic metal specie is the free hydrated metal ion. The toxicity of metals depends especially on their chemical forms rather than their total metal content. The present study focuses on Qaraaoun Reservoir, Lebanon. Earlier studies focused only on total metal concentrations in sediment and water. The objective of this study was to determine metal speciation (Fe, Cr, Ni, Zn, Cu, Pb, Cd) in the (operationally defined) sediment chemical fractions and metal speciation in reservoir water. This would reflect on metal bioavailability and toxicity. Water samples and bed sediments were collected from nine sites during the dry season and a sequential chemical fraction scheme was applied to the

Aluminum is known as a neurotoxic metal ion in experimental animals as well as in humans. The present study was carried out to determine whether and how the physicochemical properties of the metal coordination sphere (metal speciation) can influence the differentiation of murine neuroblastoma cells as has been observed previously in this laboratory (1). Results herein reported indicate that while the aluminum lipophilic species--particularly aluminum acetylacetonate (Alacac3) and aluminum maltolate (Almalt3), both hydrolitically stable and differently lipophilic--are both rather cytotoxic, metal hydrophilic species show different neuritogenic properties indicating the ability of AI(III), when diversely coordinated, to produce different biological effects.