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PubMed2004

Manganese speciation using capillary electrophoresis-ICP-mass spectrometry.

Manganese is a trace element known to activate many enzymes involved in metabolic processes and it shows protective function against oxidative stress. On the other hand, increased Mn levels are known for damaging the central nervous system, resulting in motoric abnormalities and psychic disorder. Such additional Mn exposure can cause an "Mn overflow" in the liver, accompanied by production of specific (labile) Mn transporters (Mn-species). The speciation of these Mn-compounds is still unknown but they are believed targeting the brain. The aim of this paper was to develop a speciation method for manganese species in liver extracts, which allows to speciate the compounds quickly and with minimal risk of species alteration. Capillary electrophoresis (CE)-inductively coupled plasma mass spectrometry (ICP-MS) offers a valuable tool as analytes are not in contact to a stationary phase which probably affects species stability. Separation usually is fast and ICP-MS detection is element specific and sensitive. The paper describes the set-up and optimization of the hyphenated technique, optimization of separation according to pH and finally the Mn speciation of a liver extract. Several Mn species were found, such as arginase, Mn-transferrine, Mn-albumine and some more. The detection limit of the method was determined at 1.1 microg Mn/L independent on the species.

Electrophoresis, Capillary↗
PubMed2005

Organotin speciation in environmental matrices by automated on-line hydride generation-programmed temperature vaporization-capillary gas chromatography-mass spectrometry detection.

In the present contribution, a new automated on-line hydride generation methodology was developed for dibutyltin and tributyltin speciation at the trace level, using a programmable temperature-vaporizing inlet followed by capillary gas chromatography coupled to mass spectrometry in the selected ion-monitoring mode acquisition (PTV-GC/MS(SIM)). The methodology involves a sequence defined by two running methods, the first one configured for hydride generation with sodium tetrahydroborate as derivatising agent and the second configured for speciation purposes, using a conventional autosampler and data acquisition controlled by the instrument's software. From the method-development experiments, it had been established that injector configuration has a great effect on the speciation of the actual methodology, particularly, the initial inlet temperature (-20 degrees C; He: 150 ml/min), injection volume (2 microl) and solvent characteristics using the solvent venting mode. Under optimized conditions, a remarkable instrumental performance including very good precision (RSD < 4%), excellent linear dynamic range (up to 50 microg/ml) and limits of detection of 0.12 microg/ml and 9 ng/ml, were obtained for dibutyltin and tributyltin, respectively. The feasibility of the present methodology was validated through assays upon in-house spiked water (2 ng/ml) and a certified reference sediment matrix (Community Bureau of Reference, CRM 462, Nr. 330 dibutyltin: 68+/-12 ng/g; tributyltin: 54+/-15 ng/g on dry mass basis), using liquid-liquid extraction (LLE) and solid-phase extraction (SPE) sample enrichment and multiple injections (2 x 5 microl) for sensitivity enhancement. The methodology evidenced high reproducibility, is easy to work-up, sensitive and showed to be a suitable alternative to replace the currently dedicated analytical systems for organotin speciation in environmental matrices at the trace level.

Automation↗
PubMed2006

Anion-exchange high performance liquid chromatography hyphenated to inductively coupled plasma-isotope dilution-time-of-flight mass spectrometry for speciation analysis of metal complexes with metallothionein isoforms in gibel carp (Carassius auratus gibelio) exposed to environmental metal pollution.

The capability of post-column isotope dilution (ID) combined with anion-exchange HPLC-ICP-time-of-flight (TOF)-MS was for the first time investigated for environmental quality assessment through metal speciation analysis of metallothionein (MT) isoforms in cytosols of gibel carp (Carassius auratus gibelio), used as biomarkers for environmental metal exposure. A full spectral scanning of the biological sample (with 50 microl injection volume) using ICP-TOF-MS in transient mode allowed fast multi-isotope screening of cytosolic metal-containing fractions and to investigate the presence of matrix-induced interferences. The MT cytosolic fraction of liver and kidney of the carp, sampled at three different sampling sites in Belgium, was partially purified using size-exclusion (SE) HPLC. Quantification of the elements Cd (toxic) and Zn and Cu (essential) associated with MT isoforms in this fraction was addressed using an hybrid approach based on post-column addition of the enriched isotopes 65Cu, 67Zn, 106Cd and monitoring on-line the isotope ratios 63Cu/65Cu, 64Zn/67Zn and 114Cd/106Cd by ICP-MS with a time of flight instrument, which was coupled to anion-exchange HPLC. With this separation method, baseline separation of up to five MT isoforms, which is required for quantitative metal speciation by HPLC-ICP-IDMS, was achieved within a run of 15 min. The MT fraction of the cytosols was also analysed for the total metal content using IDMS with size-exclusion HPLC-ICP-MS and species-unspecific calibration. Results showed significant differences between speciation results and total MT concentrations of control fish and fish from the most contaminated sampling sites, revealing the potential of gibel carp MT for sequestering excess intracellular free-ions (essential and toxic elements) and for its protection against metal toxicity. Preferences for metal sequestration of metal complexes with MT isoforms were also found to be tissue-specific: excess of Cd was found preferably bound to a major MT isoform (tR = 8.0 min) in kidney, whereas excess intracellular Zn appeared to be mostly sequestered by four MT isoforms (tR=7.3, 8.0, 12.2 and 14.4 min) in liver, the MT form with tR = 8.0 min being the main Zn scavenger form. Such kind of quantitative speciation information on the preferences of MT isoforms in different fish organs for sequestering heavy metals, reported here for the first time, is important to elucidate the role of isoform-specific induction of vertebrate fish MT in metal detoxification and the use of MT as biomarker.

Animals↗
PubMed2006

Current perspectives in analyte extraction strategies for tin and arsenic speciation.

Nowadays, reliable and robust detectors can be considered standard laboratory instrumentation, which, for most of the elements provide quantitation limits in the lower ng/g range. Despite these advances in detector technology, sample preparation is by far the most important error source in modern analytical method development and can be judged as the "Achilles' heel" of any analytical process regarding reliability of the obtained results and time consumption. The aim of the present review is to highlight modern trends for tin and arsenic speciation, as these analytes can be considered as models for challenges in modern method development in this field. First background information, legislative aspects and current needs are elucidated. Then the role of sample treatment within the process of method development in speciation is discussed, followed by a presentation of modern extraction techniques, matching the requirements for arsenic and tin speciation analysis: to provide mild conditions in order to ensure species preservation, to improve species recovery, to enhance sample throughput and to be suitable for hyphenation with chromatographic separation systems. The review includes applications on tin and arsenic speciation, covering the period of 2001-2006.

Analytic Sample Preparation Methods↗
PubMed2006

Nickel speciation in the presence of different sources and fractions of dissolved organic matter.

This study evaluated nickel (Ni) speciation in the presence of different fractions (humic acid (HA), fulvic acid (FA)) and sources (natural sediment, Suwannee River, peat moss) of dissolved organic matter (DOM) at Ni concentrations toxicologically relevant to the freshwater amphipod, Hyalella azteca. The free Ni ion, Ni(2+), was measured in reconstituted water (with or without DOM) using a miniaturized ion-exchange technique (IET). Ni speciation from these experiments was compared to calculated results obtained from equilibrium modelling (WHAM, Model VI). While it is known that Ni will complex with DOM, it was found that under acutely toxic Ni exposure concentrations ([Ni(Total)]=5mg/L, or 85.1 microM) representative surface-water DOC concentrations ( approximately 10mg/L) played little or no role in Ni speciation. Conversely, at sublethal Ni exposure concentrations ([Ni(Total)]=0.2 and 0.5 microg/L, or 3.4 and 8.51 microM, respectively) DOM significantly affected Ni speciation with [Ni(2+)] decreasing with increasing concentration of DOM. It was found that for similar concentrations of DOC (same fraction, different sources), the measured Ni(2+) concentrations were reduced (relative to the control), but similar to one another. Conversely, at similar DOC concentrations, the HA fraction reduced Ni(2+) levels to a greater extent than the associated FA fraction. Overall, this study provides proof of principle that Suwannee River and peat humic substances are suitable analogues for natural sediment pore-water DOM when evaluating Ni bioavailability in freshwater.

Amphipoda↗
PubMed2005

Biodegradation and speciation of residual SS-ethylenediaminedisuccinic acid (EDDS) in soil solution left after soil washing.

This paper aims to investigate the degradation and speciation of EDDS-complexes (SS-ethylenediaminedisuccinic acid) in soil following soil washing. The changes in soil solution metal and EDDS concentrations were investigated for three polluted soils. EDDS was degraded after a lag phase of 7-11 days with a half-life of 4.18-5.60 days. No influence of EDDS-speciation on the reaction was observed. The decrease in EDDS resulted in a corresponding decrease in solubilized metals. Changes in EDDS speciation can be related to (1) initial composition of the soil, (2) temporarily anoxic conditions in the soil slurry after soil washing, (3) exchange of EDDS complexes with Cu even in soils without elevated Cu and (4) formation of NiEDDS. Dissolved organic matter is important for metal speciation at low EDDS concentrations. Our results show that even in polluted soils EDDS is degraded from a level of several hundred micromoles to below 1 microM within 50 days.

Chelating Agents↗
PubMed2006

Pb speciation versus TCLP release in army firing range soils.

A series of soil parameter and mineralogical investigative techniques were applied to assess the Pb speciation in four US Army firing range soils that presented significantly different Pb leaching regimes and soil characteristics. Soil gradation tests were complemented by total chemical analyses, X-ray powder diffraction (XRPD), Rietveld quantification, optical microscopy and scanning electron microscopy (SEM) analyses. The bulk geotechnical, mineralogical and chemical analyses pointed to two possible Pb retention mechanisms: precipitation as lead carbonate and sorption in the case of fine-grained soils. Lead speciation and mobility was further investigated by the toxicity characteristic leaching procedure (TCLP) and sequential extraction test (SET). As the TCLP Pb concentrations did not necessarily reflect the total Pb analysis of the soils, the Pb leachability ratio (TCLP/total) was found to be controlled by soil mineralogy and its response to changes in system pH. Geochemical modeling, using Visual MINTEQ, was employed to evaluate the mechanisms that controlled the observed TCLP Pb leaching behavior. It was found that lead carbonate precipitation/dissolution reactions controlled Pb TCLP leachability in all soils, while sorptive phenomena did not seem to play a role even in the case of fine-grained soils. More specifically, TCLP Pb leachability was controlled by the pH, the available Pb and the available carbonate in solution. This indicates that geochemical modeling strongly complimented TCLP Pb analyses. Thus, geochemical modeling is an important assessment tool to evaluate the magnitude of site-specific Pb-related environmental problems in firing range soils. Carbonation reactions, involving metallic Pb, that occur during the SET obscure its ability to reliably ascertain Pb speciation. More specifically, SET lumps the extractable Pb into predetermined phase categories that may not be truly representative of the actual soil mineralogy or dominant forms of Pb in the soil. A thorough geotechnical, mineralogical and chemical investigation of firing range soils, complemented by geochemical modeling, was therefore found to be a more reliable approach to evaluate Pb speciation and TCLP release in firing range soils.

Lead↗
PubMed2005

Aluminium speciation in the Vienne river on its upstream catchment (Limousin region, France).

The aim of this study was to investigate the speciation of aluminium in the river Vienne on its upstream catchment (Limousin region, France) over a period of seven years (May 1998-September 2004) in order to assess harmful effects on aquatic life. Two sampling points were selected: the first at 4 km from the spring (Peyrelevade), and the second one at 89 km from the spring (Royères). The aluminium speciation was computed with Mineql+ 4.5 speciation software. Organic matter and phosphorous play a major role in aluminium speciation. If we consider the free aluminium ion (Al3+) as being the only toxic form of aluminium, the concentrations of toxic forms recorded at Peyrelevade and Royères were always below the toxic values for fish. However, if the sum of the concentrations of Al3+, Al(OH)2+, Al(OH)2+ and Al(OH)4- is taken into consideration, the concentration of aluminium recorded may have adverse effects on aquatic life in the upstream catchment of the river Vienne. Al(OH)4- is the major contributor to the concentration in toxic aluminium recorded. In general, Al(OH)4- forms appears in water during the summer with water alkalinisation due to an increase in photosynthetic activities.

Journal Article↗
PubMed2004

Speciation of arsenic in biological samples.

Speciation of arsenicals in biological samples is an essential tool to gain insight into its distribution in tissues and its species-specific toxicity to target organs. Biological samples (urine, hair, fingernail) examined in the present study were collected from 41 people of West Bengal, India, who were drinking arsenic (As)-contaminated water, whereas 25 blood and urine samples were collected from a population who stopped drinking As contaminated water 2 years before the blood collection. Speciation of arsenicals in urine, water-methanol extract of freeze-dried red blood cells (RBCs), trichloroacetic acid treated plasma, and water extract of hair and fingernail was carried out by high-performance liquid chromatography (HPLC)-inductively coupled argon plasma mass spectrometry (ICP MS). Urine contained arsenobetaine (AsB, 1.0%), arsenite (iAs(III), 11.3), arsenate (iAs(V), 10.1), monomethylarsonous acid (MMA(III), 6.6), monomethylarsonic acid (MMA(V), 10.5), dimethylarsinous acid (DMA(III), 13.0), and dimethylarsinic acid (DMA(V), 47.5); fingernail contained iAs(III) (62.4%), iAs(V) (20.2), MMA(V) (5.7), DMA(III) (8.9), and DMA(V) (2.8); hair contained iAs(III) (58.9%), iAs(V) (34.8), MMA(V) (2.9), and DMA(V) (3.4); RBCs contained AsB (22.5%) and DMA(V) (77.5); and blood plasma contained AsB (16.7%), iAs(III) (21.1), MMA(V) (27.1), and DMA(V) (35.1). MMA(III), DMA(III), and iAs(V) were not found in any plasma and RBCs samples, but urine contained all of them. Arsenic in urine, fingernails, and hair are positively correlated with water As, suggesting that any of these measurements could be considered as a biomarker to As exposure. Status of urine and exogenous contamination of hair urgently need speciation of As in these samples, but speciation of As in nail is related to its total As (tAs) concentration. Therefore, total As concentrations of nails could be considered as biomarker to As exposure in the endemic areas.

Arsenicals↗
PubMed2005

Speciation through the learning of habitat features.

Learning of environmental features can influence both mating behaviour and the location where young are produced. This may lead to speciation in three steps: (i) colonization of a new habitat, (ii) genetic divergence of the two groups by adaptation to the habitats, and (iii) a decrease of genetic mixing between the lineages (similar to reinforcement). In a previous paper we showed that steps (i) and (ii) occur readily for a wide range of fixed mating and habitat preferences. Here, we study whether this can ultimately lead to speciation through selective changes in these preferences. We show that this indeed occurs, and, furthermore, it is very general: for a large class of models there is selection toward producing young more frequently in the natal habitat. Once habitat preference is strong, there is selection toward stronger assortative mating. Even when steps (i) and (ii) initially fail, genetic divergence may succeed at a later evolutionary stage, after which a decrease of genetic mixing completes speciation. Our results show that speciation by the learning of habitat features is an extremely effective mechanism.

Animals↗
PubMed2004

Speciation by host-switching in pinyon Cinara (Insecta: Hemiptera: Aphididae).

Parasite-host cospeciation has received much attention as an important mechanism in the diversification of phytophagous insects. However, studies have shown that for certain taxa, it is not host fidelity but host-switching that plays the critical role in speciation. Cinara are aphids (Insecta: Hemiptera: Aphididae: Lachninae) that feed exclusively on the woody parts of conifers of the Cupressaceae and Pinaceae. They are unusual aphids because most Pinaceae play host to several species of Cinara. The aphids show relatively strong host fidelity, and as a consequence historically have been treated based on the taxonomy of their hosts. The historical paradigm of aphid evolution implies that Cinara species have radiated to different parts of the same host species and/or speciated with their host. Using mitochondrial cytochrome oxidase 1 and nuclear elongation factor 1-alpha DNA sequences, we performed molecular phylogenetic analysis of Cinara species, concentrating on those associated with pinyon pines in the southwestern USA. We determined that switching hosts has played a key role in the speciation of the genus, reflected in the polyphyly of pinyon-feeding Cinara. Furthermore, species sharing a common feeding site on different hosts were more closely related to each other than to those sharing the same host but at different feeding sites, suggesting that feeding site fidelity plays a more important role in speciation than does host fidelity in general. This study also elucidated the primary taxonomy of various species: it suggested that Cinara rustica Hottes is a junior synonym of C. edulis (Wilson) and that C. wahtolca Hottes represents two species on the two different pinyon pine species, Pinus edulis Englem. and P. monophylla Torr. & Frem.

Animals↗
PubMed2005

Testing hypotheses of speciation timing in Dicamptodon copei and Dicamptodon aterrimus (Caudata: Dicamptodontidae).

Giant salamanders of the genus Dicamptodon are members of the mesic forest ecosystem that occurs in the Pacific Northwest of North America. We estimate the phylogeny of the genus to test several hypotheses concerning speciation and the origin of current species distributions. Specifically, we test competing a priori hypotheses of dispersal and vicariance to explain the disjunct inland distribution of the Idaho giant salamander (D. aterrimus) and to test the hypothesis of Pleistocene speciation of Cope's giant salamander (D. copei) using Bayesian hypothesis testing. We determined that available outgroups were too divergent to root the phylogeny effectively, and we calculated Bayesian posterior probabilities for each of the 15 possible root placements for this four-taxon group. This analysis placed the root on the branch leading to D. aterrimus, indicating that current distribution and speciation of D. aterrimus fit the ancient vicariance hypothesis and are attributable to the orogeny of the Cascade Mountains rather than recent inland dispersal. Furthermore, test results indicate that D. copei is distantly related to other coastal lineages and likely originated much earlier than the Pleistocene. These results suggest that speciation within the genus is attributable to ancient geologic events, while more recent Pleistocene glaciation has shaped genetic variation and distributions within the extant species.

Animals↗
PubMed2005

Interspecific phylogeography of the Stator limbatus species complex: the geographic context of speciation and specialization.

Diversification in phytophagous insects is often attributed to a propensity toward specialization and to a tendency for speciation to be associated with host-shifts. Phylogenetic analysis revealed a sister relationship between the generalist Stator limbatus and the specialist host-shifted Stator beali, providing a system to examine the genealogical and geographic origins of the main processes involved in this diversification: host-shifts, specialization, and reproductive isolation. We examine the interspecific phylogeographic relationships between these species using mitochondrial DNA sequence data. S. beali is derived within S. limbatus, rendering the latter paraphyletic and suggesting a budding process of speciation. The inherent polarity in this genealogical pattern indicates that the specialist habit, clumping oviposition behavior, and distinct genitalia of S. beali are all derived from the ancestral S. limbatus. The phylogeography of S. limbatus also shows strong geographic structure with divergences corresponding to known biogeographic boundaries, indicating that this evolutionary signal has not been erased by the vagaries of history. However, the derivation of S. beali and the evolution of reproductive isolation between the two species does not correspond to these known biogeographic boundaries, as S. beali and its sister clade of S. limbatus are restricted to the same geographic province. The geographic proximity of diversification combined with a divergence time estimated at the beginning of the Pleistocene indicates that speciation likely occurred very rapidly, although further genetic and ecological work is necessary to examine the mode of speciation. This study provides the historical context for ongoing evolutionary, ecological, and quantitative genetic research on the divergence in diet breadth between these species.

Animals↗
PubMed1998

Al and Si: their speciation, distribution, and toxicity.

OBJECTIVES: In dialysis patients both aluminum (AI) and silicon (Si) may accumulate. Whereas the toxic effects of AI within this population are clearly established, little is known on the role of Si in the development/protection of particular dialysis-related diseases. A clear insight in the protein binding and speciation of trace elements is important to better understand the mechanisms underlying their toxicity/essentiality. Research in this field however is complex and often prone to analytical difficulties and inaccuracies. DESIGN AND METHODS: In the first part of this review techniques used for speciation studies of AI and Si in biological fluids are discussed. Notwithstanding recent technical advances (a) extraneous metal contamination, (b) unrecognized aspecific binding of metals to proteins, and (c) unwanted interactions with separation equipment such as chromatography columns and ultrafiltration membranes remain important pitfalls and often lead to erroneous conclusions. The factors that determine the speciation of AI and Si and their ultimate tissue distribution and toxicity are dealt with in the second part. Here, experimental data obtained with various speciation techniques are linked to in vivo data on the tissue distribution, localization/toxicity of both elements. CONCLUSIONS: A model in which the AI tissue distribution/toxicity is mediated by either its citrate or transferrin bound form is proposed.

Aluminum↗
PubMed2002

Speciation of inorganic arsenic in some bottled Slovene mineral waters using HPLC-hGAFS and selective coprecipitation combined with FI-HGAFS.

Five bottled mineral waters from the Radenska and the Rogaska springs (Slovenia) were analyzed for their inorganic arsenic species As(III) and As(V) using existing speciation procedures. Both a hyphenated technique (HPLC-HGAFS) and a more conventional technique based on selective coprecipitation of As(III) with dibenzyldithiocarbamate prior to arsenic analysis (FI-HGAFS) were used. The techniques yielded data which were not significantly different on the 5% level, with HPLC-HGAFS to be the least sensitive (DL 1 microgl(-1)) and the selective coprecipitation technique to be suitable for sub pgl(-1) levels (DL 0.05 microgl(-1)). The latter technique showed recoveries of 96.4 +/- 0.4%. In none of the mineral waters As(III) was found whereas the Radenska minerals waters (vrelec Radin vrelec Miral, Kraljevi Vrelec) contained little As(V) (only detectable with the selective coprecipitation technique) and the Rogaska mineral waters (Tempel, Donat Mg) (very) high As(V) concentrations. Rogaska, Donat Mg even had an average As(V) concentration around the permittable level of 50 microgl(-1) for arsenic in minerals waters, viz. 47.8 microg l(-1). Speciation calculations based on pH and Eh and taking into account the ionic strength gave a similar speciation pattern then speciation analysis with additional information about the degree of protonation of As(III) and As(V).

Arsenic↗
PubMed2000

Speciation of aluminium in forest soil extracts by size exclusion chromatography with UV and ICP-AES detection and cation exchange fast protein liquid chromatography with ETAAS detection.

Aluminium speciation was studied in forest soil extracts by size exclusion chromatography (SE) with UV and inductively coupled plasma-atomic emission spectrometric (ICP-AES) detection and cation exchange fast protein liquid chromatography (FPLC) with ETAAS detection. Size exclusion chromatography was performed on a Superdex HR75 10/30 column. Isocratic clution with 0.15 mol dm(-3) NaCl in TRIS-HCl buffer (pH = 5.5) was applied over 100 min at a flow rate of 0.35 cm(3) min(-1). The chromatographic run was followed at 278 nm and separated Al species also determined 'off line' in 0.875 cm3 fractions by ICP-AES. The analytical procedure enabled speciation of high molecular weight Al complexes. Cation exchange FPLC was performed on a Mono S HR 5/5 column. Aqueous 8 mol dm(-3) NH4NO3 linear gradient elution was applied over 10 min at a flow rate of 1 cm3 min(-1). Separated Al species were collected in 0.5 cm3 fractions and Al determined 'off line' by ETAAS. The analytical procedure enabled speciation of some positively charged monomeric Al species. Negatively charged species were eluted with the solvent front. The combination of the two analytical techniques was successfully employed in speciation of Al in forest soil extracts. Water was used as an extracting solution. It was found experimentally that 80-95% of Al in aqueous extracts of forest soils exists in monomeric Al forms. Water soluble Al (30-40%) is bound to high molecular weight complexes with humic substances. The remaining monomeric Al in the low molecular weight fraction exists as AIF2+, Al-oxalate and Al-citrate species.

Aluminum↗
PubMed2001

The importance of ligand speciation in environmental research: a case study.

The speciations of EDTA and DTPA in process, waste and river waters are modelled and simulated, specifically to the mode of occurrence in the pulp and paper mill effluents and subsequently in receiving waters. Due to relatively short residence times in bleaching process and waste water treatment and slow exchange kinetics, it is expected that the thermodynamic equilibrium is not necessarily reached. Therefore, the initial speciation plays a key role. As such, the simulations have been extended to the process waters of the pulp and paper industry taking into account estimated average conditions. The results reveal that the main species are; Mn and Ca complexes of EDTA and DTPA in pulp mill process waters; Fe(III) and Mn complexes of EDTA and DTPA in waste waters; Fe(III) and Zn complexes of EDTA and DTPA in receiving waters. It is also shown how the increasing concentration of complexing agents effects the speciation. Alkaline earth metal chelation plays a significant role in the speciation of EDTA and DTPA when there is a noticeable molar excess of complexing agents compared with transition metals.

Chelating Agents↗
PubMed2003

Possible method of aluminium speciation in forest soils.

Labile Al forms and species can be a threat in acid soils due to their potential toxicity to plants. However, there is no universally accepted extraction method. Several extraction reagents for Al release from soil have been tested. KCl (0.5 or 1 M) is recommended for extraction of exchangeable Al, while 0.5 or 0.3 M CuCl(2) is suggested for extraction of 'weakly organically bound Al'. Both 0.1 and 0.05 M Na(4)P(2)O(7) are shown to be suitable for the extraction of 'total organically bound Al'. These extractions are relatively simple, robust, and applicable to different soils and soil horizons. In the second part of the paper, detailed speciation of exchangeable soil Al by means of an HPLC instrument equipped with an ion column (IC) is presented. An experimental set-up is described and tested on a set of samples. Interpretation of the speciation results is proposed, based on the separation of Al ions and Al complexes according to their charge. Speciation is shown to be dependent mainly on soil pH and organic matter quality. A general scheme of Al fractionation and speciation in soil is proposed.

Aluminum↗