The on-line coupling of capillary electrophoresis (CE) and electrospray ionization mass spectrometry (ESI-MS) for speciation analysis is entailed with a number of difficulties due to technical and methodical reasons. The optimization of... more
The on-line coupling of capillary electrophoresis (CE) and electrospray ionization mass spectrometry (ESI-MS) for speciation analysis is entailed with a number of difficulties due to technical and methodical reasons. The optimization of several parameters in order to provide stable operating conditions are illustrated in detail: the positioning of the CE capillary, the CE buffer system, the concentration sensitivity and the detection mode. Additionally, off-line ESI-MS investigations of metal species with different stabilities were carried out in order to assess the influence of the ESI-process on the analyzed samples. It turned out, that metal ions and weak metal complexes may undergo gas-phase ligand replacement and intramolecular charge transfer reactions. Therefore, the structure of species will be altered. Covalent organometallic compounds and strong metal complexes, on the other hand, will not be altered and will be detected as charged molecular ions and/or as charged ion-solv...
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The high resolution potential of capillary electrophoresis (CE) makes CE techniques valuable for separations of selenium species, both, organic and inorganic. Such effective separations of charged species are possible using the CZE mode... more
The high resolution potential of capillary electrophoresis (CE) makes CE techniques valuable for separations of selenium species, both, organic and inorganic. Such effective separations of charged species are possible using the CZE mode (Z = zone). Very powerful separations can be achieved by capillary isoelectric focusing, when analysing molecules showing different isoelectric points (pI) values, such as organic Se species. Inductively coupled plasma (ICP) MS is an element-specific multi-element detector, providing extremely low detection limits. The combination of CE with ICP-MS promises a powerful tool for metal speciation. Therefore, an online hyphenation of CE [using capillary zone electrophoresis (CZE) or capillary isoelectric focusing (cITP) model] with ICP-MS, which was developed earlier, was modified and applied to selenium speciation. Separation was differentiated from the detection step during hyphenation. This resulted in short separation times (10 min) and a subsequent ...
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Arsenic and inorganic arsenic compounds (with the exception of arsenic hydride): BAT value documentation (2016) The German Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area has re-evaluated the... more
Arsenic and inorganic arsenic compounds (with the exception of arsenic hydride): BAT value documentation (2016) The German Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area has re-evaluated the “Biologischer Arbeitsstoff-Referenzwert” (BAR) as well as the exposure equivalents for carcinogenic substances (EKA) for arsenic and inorganic arsenic compounds (with the exception of arsenic hydride). Available publications are described in detail. The quantification of the various arsenic species is essential to understand the hazardous potential of the arsenic compounds which differ highly in their toxicity. Therefore the BAR for the sum of different arsenic species is no longer valid. Considering published data from occupational not exposed persons, new BAR for the arsenic species of 0.5 μg/l urine for arsenic (+III), 0.5 μg/l urine for arsenic (+V), 2 μg/l urine for monomethylarsonic acid (MMA) and 10 μg/l urine for dimethylarsinic acid (DMA) were ...
AIM Extracorporeal shockwave application (ESWA) is in clinical use to promote disintegration of symptomatic calcifications of the rotator cuff of the shoulder. However, disintegration of roentgenologically comparable calcifications of the... more
AIM Extracorporeal shockwave application (ESWA) is in clinical use to promote disintegration of symptomatic calcifications of the rotator cuff of the shoulder. However, disintegration of roentgenologically comparable calcifications of the rotator cuff is not always successfull. It is known from urologic stone lithotripsy that the susceptibility for disintegration of stone-like concrements depends on their mineral content. Therefore, in the present investigation the relative contents of calcium and phosphorus in rotator cuff calcifications were determined. METHOD 39 surgically removed rotator cuff calcifications were analyzed by means of atomic emission spectrometry. RESULTS The relative content of calcium of the rotator cuff calcifications was found to be 22.3 % +/- 5.7 % (mean +/- SD; 6.8 % - 32.4 %), that of phosphorus as 10.5 % +/- 2.4 % (2.7 % - 14.4 %). The data neither depend on the gender of the patients nor on their age at the time point of surgical removal of the rotator cuff calcifications. CONCLUSION Roentgenologically comparable calcifications of the rotator cuff demonstrated distinct individual differences concerning the relative contents of calcium and phosphorus. The present results may serve as the first indication that the susceptibility of rotator cuff calcifications for disintegration may depend on their relative contents of calcium and phosphorus.
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ABSTRACT
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Extracorporeal shockwave application (ESWA) is in clinical use to promote disintegration of symptomatic calcifications of the rotator cuff of the shoulder. However, disintegration of roentgenologically comparable calcifications of the... more
Extracorporeal shockwave application (ESWA) is in clinical use to promote disintegration of symptomatic calcifications of the rotator cuff of the shoulder. However, disintegration of roentgenologically comparable calcifications of the rotator cuff is not always successfull. It is known from urologic stone lithotripsy that the susceptibility for disintegration of stone-like concrements depends on their mineral content. Therefore, in the present investigation the relative contents of calcium and phosphorus in rotator cuff calcifications were determined. 39 surgically removed rotator cuff calcifications were analyzed by means of atomic emission spectrometry. The relative content of calcium of the rotator cuff calcifications was found to be 22.3 % +/- 5.7 % (mean +/- SD; 6.8 % - 32.4 %), that of phosphorus as 10.5 % +/- 2.4 % (2.7 % - 14.4 %). The data neither depend on the gender of the patients nor on their age at the time point of surgical removal of the rotator cuff calcifications. ...
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This chapter describes strategies and methods for speciation investigations using mainly hyphenated techniques of liquid chromatography with element-specific detection. Complete speciation schemes are discussed, starting with the sources... more
This chapter describes strategies and methods for speciation investigations using mainly hyphenated techniques of liquid chromatography with element-specific detection. Complete speciation schemes are discussed, starting with the sources of error and problem solutions for sampling and ending with more dimensional, orthogonally chosen coupled-speciation techniques for quality controlled results. The focus in each section is directed to the specific problems being faced with respect to speciation analysis: species stability during sample storage and processing, the choice of the most suitable speciation approaches (direct methods or hyphenated techniques), species separation techniques ranging from ultrafiltration to various forms of liquid chromatography, and capillary electrophoresis, discussing their advantageous as well as their (undesired) potential of changing species. Further, problems in interfacing to element-specific detectors, mainly plasma-based detectors like ICP OES and ICP MS are reported. The chapter concludes with aspects of quality control, including sources of errors and strategies to overcome such problems, such as orthogonal speciation concepts.
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Following the end of the Kosovo conflict, in June 1999, a study was instigated to evaluate whether there was a cause for concern of health risk from depleted uranium (DU) to German peacekeeping personnel serving in the Balkans. In... more
Following the end of the Kosovo conflict, in June 1999, a study was instigated to evaluate whether there was a cause for concern of health risk from depleted uranium (DU) to German peacekeeping personnel serving in the Balkans. In addition, the investigations were extended to residents of Kosovo and southern Serbia, who lived in areas where DU ammunitions were deployed. In order to assess a possible DU intake, both the urinary uranium excretion of volunteer residents and water samples were collected and analysed using inductively coupled plasma-mass spectrometry (ICP-MS). More than 1300 urine samples from peacekeeping personnel and unexposed controls of different genders and age were analysed to determine uranium excretion parameters. The urine measurements for 113 unexposed subjects revealed a daily uranium excretion rate with a geometric mean of 13.9 ng/d (geometric standard deviation (GSD)=2.17). The analysis of 1228 urine samples from the peacekeeping personnel resulted in a geometric mean of 12.8 ng/d (GSD=2.60). It follows that both unexposed controls and peacekeeping personnel excreted similar amounts of uranium. Inter-subject variation in uranium excretion was high and no significant age-specific differences were found. The second part of the study monitored 24 h urine samples provided by selected residents of Kosovo and adjacent regions of Serbia compared to controls from Munich, Germany. Total uranium and isotope ratios were measured in order to determine DU content. (235)U/(238)U ratios were within +/-0.3% of the natural value, and (236)U/(238)U was less than 2 x 10(-7), indicating no significant DU in any of the urine samples provided, despite total uranium excretion being relatively high in some cases. Measurements of ground and tap water samples from regions where DU munitions were deployed did not show any contamination with DU, except in one sample. It is concluded that both peacekeeping personnel and residents serving or living in the Balkans, respectively, were not exposed to significant amounts of DU.
Research Interests: Epidemiology, Mass Spectrometry, Environmental Monitoring, Public Health, Comparative Study, and 15 moreMultidisciplinary, Germany, Humans, Female, Male, Inductively Coupled Plasma Mass Spectrometry, Aged, Middle Aged, Adult, Occupational Exposure, Health risk, Depleted Uranium, Military Personnel, excretion, and Geometric mean
The speciation of trace element species in solid matrices like liver samples is still problematic due to two reasons. On the one hand direct methods with sufficient selectivity and sensitivity are currently not available. Therefore... more
The speciation of trace element species in solid matrices like liver samples is still problematic due to two reasons. On the one hand direct methods with sufficient selectivity and sensitivity are currently not available. Therefore extraction procedures have to be applied which are often problematic in respect to species stability. On the other hand there are no reference materials with known amounts of metal proteins like metallothionein-isoforms (MT) and superoxide dismutase (SOD) for quality control. So the aim of this study was to develop and optimise procedures for the species-preserving extraction of the model compounds MT and SOD from liver samples. Spiking experiments were performed to overcome the lack of appropriate reference materials. In a first step the stability of the model species without liver matrix was investigated by the variation of several extraction parameters. The extractant and exposure to ultrasonic energy especially had a great influence on the recovery of the species while temperature, buffer concentration and atmospheric conditions were less critical. In a second step spiked liver samples were extracted with a selection of procedures taken from the literature. Most of these methods provided recoveries between 70% and 100%. Additionally the buffer concentration and the extractant-to-liver ratio were varied for optimisation. The metal balance of an extraction showed recoveries of 81% for Cd, 94% for Cu and 87% for Zn.
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Background/Objectives:The apparent widespread extent of zinc (Zn) deficiency in developing countries and the efficacy of oral Zn supplements as an adjunct to oral rehydration therapy make oral Zn supplementation an increasingly important... more
Background/Objectives:The apparent widespread extent of zinc (Zn) deficiency in developing countries and the efficacy of oral Zn supplements as an adjunct to oral rehydration therapy make oral Zn supplementation an increasingly important modality in clinical medicine and public health. In this study we aimed to compare the relative bioavailability of oral doses of 30 mg of Zn in two dosing formsSubjects/Methods:In total, 10 healthy male volunteers ingested oral Zn doses with 200 ml plain water at about 0830 hours in the fasting state on two occasions, once as 30 mg of Zn in an aqueous solution of reagent grade zinc sulfate (ZnSO4) and another time as 1.5 NutriSet Zn tablets (Nutriset, Malaunay, France); on a third occasion, only plain water was consumed. Venous blood specimens were collected at baseline, 60, 120, 180 and 240 min after ingestion and the plasma Zn was measured for each sample.Results:The relative bioavailability of oral Zn from a commonly used, tableted (NutriSet) for...
Research Interests: Metabolism, Developing Countries, Adolescent, Comparative Study, Humans, and 15 moreGuatemala, Developing Country, Fasting, Male, Dietary Supplements, Bioavailability, Nutrient, Middle Aged, Adult, Food Sciences, Aqueous Solution, Biological Availability, Area Under the Curve, Calories, and functional outcome
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Improved living conditions have led to a steady increase in the life expectancy of humans in most countries. However, this is accompanied by an increased probability of suffering from neurodegenerative diseases like... more
Improved living conditions have led to a steady increase in the life expectancy of humans in most countries. However, this is accompanied by an increased probability of suffering from neurodegenerative diseases like Alzheimer's disease or Parkinson's disease. Unfortunately, the therapeutic possibilities for curing these diseases are very limited up to now. Many studies indicate that a variety of environmental factors contribute to the initiation and promotion of neurodegenerative diseases. For example, the role of metal exposure and disturbance of metal homeostasis in the brain is discussed in this respect. However, most studies focus on the neurological and toxicological aspects but not on a detailed characterisation of the species of the involved metals. Therefore, this review summarizes the neurotoxic effects of selected metals on humans and focuses on contributions from trace element speciation analysis with relevance to neuroscientific research. In spite of the advance in instrumentation and methodology of speciation analysis there are few applications for matrices like cerebrospinal fluid which is due to limited access to these samples and analytical challenges caused by matrix interferences, low concentrations and limited stability of many trace element species of interest. The most relevant neurotoxic metals aluminium, lead, manganese and mercury are reviewed in detail while further metals like cadmium, arsenic, bismuth and tin are briefly discussed. Current results indicate that knowledge on trace element speciation can contribute to a better understanding of the transport of metals across the neural barriers and potentially of their role in diseased human brains.
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During the second half of the 20th century it became evident that trace elements play a major role whenever biological activities, environmental chemistry, or material characteristics are discussed and the determination of trace elements... more
During the second half of the 20th century it became evident that trace elements play a major role whenever biological activities, environmental chemistry, or material characteristics are discussed and the determination of trace elements has therefore gained outstanding importance in environmental and life sciences. Elements, even when present at minimal concentrations in biological and environmental matrices, can, in fact, exert fundamental influence on ecosystems and the vital functions of organisms. The study of, for example, pathophysiological processes in the human body requires the determination of elements down to the ng kg(-1) range. However, organic and inorganic matrix compounds at much higher concentration levels in a sample than trace elements make the determination of trace elements often rather difficult.
Research Interests: Environmental Monitoring, Risk assessment, Environmental Sciences, Humans, Elements, and 11 moreCHEMICAL SCIENCES, Ecosystem, Inorganic chemicals, Reproducibility of Results, Microchemistry, Risk Assessment, Environmental Pollutants, Sensitivity and Specificity, Geologic Sediments, Organic Chemicals, and Medical and Health Sciences
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Research Interests: Speciation, Mass Spectrometry, Capillary electrophoresis, Sample Preparation, Soils, and 11 moreChemical Analysis, Sol, Contamination, Inductively Coupled Plasma Mass Spectrometry, High Performance Liquid Chromatography, EXTRACTION, High Resolution, Electrophoresis, Liquid Chromatography, Aqueous Extract, and Mass Spectrometer
Manganese (Mn) is an essential nutrient that can be toxic in excess concentrations, especially during early development stages. The mechanisms of Mn toxicity is still unclear, and little information is available regarding the role of Mn... more
Manganese (Mn) is an essential nutrient that can be toxic in excess concentrations, especially during early development stages. The mechanisms of Mn toxicity is still unclear, and little information is available regarding the role of Mn speciation and fractionation in toxicology. We aimed to investigate the toxic effects of several chemical forms of Mn in embryos of Danio rerio exposed during different development stages, between 2 and 122h post fertilization. We found a stage-specific increase of lethality associated with hatching and removal of the chorion. Mn(II), ([Mn(H2O)6](2+)) appeared to be the most toxic species to embryos exposed for 48h, and Mn(II) citrate was most toxic to embryos exposed for 72 and/or 120h. Manganese toxicity was associated with calcium disruption, manganese speciation and metal fractionation, including bioaccumulation in tissue, granule fractions, organelles and denaturated proteins.
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After high-dose-short-term exposure (usually from occupational exposure) and even more under low-dose long term exposure (mainly environmental) manganese (Mn) biomonitoring is still problematic since these exposure scenarios are not... more
After high-dose-short-term exposure (usually from occupational exposure) and even more under low-dose long term exposure (mainly environmental) manganese (Mn) biomonitoring is still problematic since these exposure scenarios are not necessarily reflected by a significant increase of total Mn in blood or serum. Usually, Mn concentrations of exposed and unexposed persons overlap and individual differentiation is often not possible. In this paper Mn speciation on a large sample size (n=180) was used in order to be able to differentiate between highly Mn-exposed or low or unexposed individuals at low total Mn concentration in serum (Mn(S)). The whole sample set consisted of three subsets from Munich, Emilia Romagna region in Italy and from Sweden. It turned out that also at low total Mn(S) concentrations a change in major Mn carriers in serum takes place from Mn-transferrin (Mn-Tf(S)) towards Mn-citrate (Mn-Cit(S)) with high statistical significance (p<0.000002). This carrier switch from Mn-Tf(S) to Mn-Cit(S) was observed between Mn(S) concentrations of 1.5μg/L to ca. 1.7μg/L. Parallel to this carrier change, for sample donors from Munich where serum and cerebrospinal fluid were available, the concentration of Mn beyond neural barriers - analysed as Mn in cerebrospinal fluid (Mn(C)) - positively correlates to Mn-Cit(S) when Mn(S) concentration was above 1.7μg/L. The correlation between Mn-Cit(S) and Mn(C) reflects the facilitated Mn transport through neural barrier by means of Mn-citrate. Regional differences in switch points from Mn-Tf(S) to Mn-Cit(S) were observed for the three sample subsets. It is currently unknown whether these differences are due to differences in location, occupation, health status or other aspects. Based on our results, Mn-Cit(S) determination was considered as a potential means for estimating the Mn load in brain and CSF, i.e., it could be used as a biomarker for Mn beyond neural barrier. For a simpler Mn-Cit(S) determination than size exclusion chromatography inductively coupled plasma mass spectrometry (SEC-ICP-MS), ultrafiltration (UF) of serum samples was tested for suitability, the latter possibly being a preferred choice for routine occupational medicine laboratories. Our results revealed that UF could be an alternative if methodical prerequisites and limitations are carefully considered. These prerequisites were determined to be a thorough cleaning procedure at a minimum Mn(S) concentration >1.5μg/L, as at lower concentrations a wide scattering of the measured concentrations in comparison to the standardized SEC-ICP-MS results were observed.
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Hip resurfacing arthroplasty is known to increase the metal ion concentration in the serum, urine and whole blood, with potentially adverse effects on the organism. However, only few data are available about the metal ion concentrations... more
Hip resurfacing arthroplasty is known to increase the metal ion concentration in the serum, urine and whole blood, with potentially adverse effects on the organism. However, only few data are available about the metal ion concentrations in erythrocytes, although they are directly exposed to the higher concentrations of the serum. The ion levels of chromium, cobalt, nickel and molybdenum in erythrocytes of 25 patients with a hip resurfacing implant were analysed with high resolution ICP-sf-MS (inductively-coupled-plasma-sector-field-mass-spectrometry). The results were compared to the ion levels in the serum and urine of the patients, and also to the ion levels of 27 control persons without an implant. Compared to the control group, ion levels in the erythrocytes of the hip resurfacing group were markedly increased for cobalt (0.10 vs. 3.26 microg/kg) and slightly for chromium (6.04 vs. 7.38 microg/kg). In contrast, ion levels in the serum of the hip resurfacing group were increased ...
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Reversed-phase liquid chromatography-inductively coupled plasma mass spectrometric hyphenation was used for iodine speciation in human serum. First investigations showed that iodine species nearly quantitatively were eluted in the void... more
Reversed-phase liquid chromatography-inductively coupled plasma mass spectrometric hyphenation was used for iodine speciation in human serum. First investigations showed that iodine species nearly quantitatively were eluted in the void volume. The result indicated that protein-linked thyroid hormones were not interacting with the stationary phase, thus being not retained. Investigations were performed about T4-TBG (thyroxin-thyroxin-binding globulin) complex generation and its retention during chromatography. It was shown that T4-TBG was not retained on the column. Therefore, a protease treatment was introduced for serum sample preparation. The analysis of "normal" sera (after protease) gave reasonable results lying in the range published in literature: I-:11; di-iodothyrosine (DIT): 2.1; mono-iodothyrosine (MIT): 1.6; reversed tri-iodothyronine (rT3): 3.9; T3: 5.9; T4: 60; each micrograms iodine per liter. The method also proved to recognize abnormalities in a pathologic ...
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Method developments are described for a iodine speciation method. This method is based on a reversed-phase liquid chromatography separation combined with inductively coupled plasma-mass spectrometry detection. Investigations are performed... more
Method developments are described for a iodine speciation method. This method is based on a reversed-phase liquid chromatography separation combined with inductively coupled plasma-mass spectrometry detection. Investigations are performed concerning the methanol tolerance/stability of inductively coupled plasma, postcolumn oxidation/volatilization of iodine from species, influences of buffer pH, and column temperature on separation efficiency. The experiments focused on six iodine species, namely iodine, monoiodothyrosine, di-iodothyrosine, tri-iodothyronine, reversed tri-iodothyronine, and thyroxin. Detection limits were determined between 0.08 and 1.5 microg/L (iodine related), dependent on species. Appplications on a urine sample were performed, showing iodide as the predominant iodine species.
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Page 1. 17 ELEMENTAL SPECIATION IN HUMAN MILK AND SUBSTITUTE FOOD FOR NEWBORNS BERNHARD MICHALKE GSF National Research Center for Environment and Health, Institute for Ecological Chemistry, Neuherberg ...
The chemical composition of particulate matter (PM) emissions from a medium-speed four-stroke marine engine, operated on both heavy fuel oil (HFO) and distillate fuel (DF), was studied under various operating conditions. PM emission... more
The chemical composition of particulate matter (PM) emissions from a medium-speed four-stroke marine engine, operated on both heavy fuel oil (HFO) and distillate fuel (DF), was studied under various operating conditions. PM emission factors for organic matter, elemental carbon (soot), inorganic species and a variety of organic compounds were determined. In addition, the molecular composition of aromatic organic matter was analyzed using a novel coupling of a thermal-optical carbon analyzer with a resonance-enhanced multiphoton ionization (REMPI) mass spectrometer. The polycyclic aromatic hydrocarbons (PAHs) were predominantly present in an alkylated form, and the composition of the aromatic organic matter in emissions clearly resembled that of fuel. The emissions of species known to be hazardous to health (PAH, Oxy-PAH, N-PAH, transition metals) were significantly higher from HFO than from DF operation, at all engine loads. In contrast, DF usage generated higher elemental carbon emissions than HFO at typical load points (50% and 75%) for marine operation. Thus, according to this study, the sulfur emission regulations that force the usage of low-sulfur distillate fuels will also substantially decrease the emissions of currently unregulated hazardous species. However, the emissions of soot may even increase if the fuel injection system is optimized for HFO operation.
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This paper compares trace element concentrations (Ca, K, Sr, Fe, Mn, Zn, Ni, Cu, Co and Cr) in 27 Sudanese medical plants determined in parallel by PIXE and ICP-OES to get information on which technique is preferable at different matrices... more
This paper compares trace element concentrations (Ca, K, Sr, Fe, Mn, Zn, Ni, Cu, Co and Cr) in 27 Sudanese medical plants determined in parallel by PIXE and ICP-OES to get information on which technique is preferable at different matrices and element concentrations. PIXE correlates well to ICP-OES for Sr, Mn, Ca, K, Zn and Fe determinations. ICP-OES seems to be the superior technique over PIXE when measuring low concentrated elements (chromium, cobalt, nickel and copper) in the medicinal plants.
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... Biological Conservation, 76: 1723. [CrossRef], [Web of Science ®], [CSA]; 3. Biney, C, Amuzu, AT, Calamari, D, Kaba, N, Mbome, IL, Nacre, H, Ochumba, PBO, Osibanjo, O, Radegonde, V and Saad, MAH. 1994. ... New York: John Wiley and... more
... Biological Conservation, 76: 1723. [CrossRef], [Web of Science ®], [CSA]; 3. Biney, C, Amuzu, AT, Calamari, D, Kaba, N, Mbome, IL, Nacre, H, Ochumba, PBO, Osibanjo, O, Radegonde, V and Saad, MAH. 1994. ... New York: John Wiley and Sons. 9. European Communities (EC). ...
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Hop growing in Bavaria requires the use of copper containing fungicides against mildew, which results in an accumulation of copper in the upper soils to concentrations of up to approximately 450 mg kg(-1). The naturally occurring... more
Hop growing in Bavaria requires the use of copper containing fungicides against mildew, which results in an accumulation of copper in the upper soils to concentrations of up to approximately 450 mg kg(-1). The naturally occurring concentration of copper is approximately 10-15 times lower. Several single and sequential extractions were performed in parallel to the determination of total copper content after aqua regia digestion. By these means copper mobility and retention, its availability to plants as well as its distribution in soils was assessed. Three agricultural soils and two soils from fallow land have been analysed to elucidate whether the copper distribution changes in relation to agricultural activities and to predict environmental risks due to the high copper concentrations. This work also focuses on the difficulties arising from different results when using single and sequential extraction procedures. EDTA, acetic acid, deionised water, ammonium nitrate and calcium chloride were used as single extractants for the functionally defined speciation. EDTA and acetic acid can be regarded as extractants for the plant-available species, while deionised water, NH4NO3 and CaCl2 only release electrostatically weakly bound metals to estimate the mobile species. The extraction behaviour of copper in these procedures is discussed with respect to the quantities extracted from the contaminated and non-contaminated soils. The studies indicate that no vertical copper translocation is observed in the ground, but the main amounts are highly available to plants. Sequential extraction procedures were used for operationally defined speciation, quantifying copper in specific soil phases. The 'five-step extraction scheme' first applied, suggested by the Standard Measurement and Testing Programme of the European Community, overestimated the amounts of copper bound to the residual phase. This was mainly due to the non-specific nature of the reagents for the organic/sulfidic and crystalline iron oxide fraction. A second reason could be the fact that this scheme was developed for sediment and not for soil analysis. Therefore, the sequential extraction has been applied using an adapted method of Zeien and Bruemmer, using different extractants for the two phases. The copper distribution was different, showing that approximately 40-50% of the total copper was bound to the organic fraction and approximately 15-30% could be extracted with the iron oxides. The amounts in the residual fraction were much lower. After comparing the results of the different extraction procedures the environmental behaviour of copper and its risks for plants, micro-organisms and groundwater is discussed in brief.