Heavy metals in the spinal cord of normal rats and of animals treated with chelating agents: a quantitative (zinc, copper, and lead) and histochemical study

The amounts of zinc, copper, and lead in the rat spinal cord were determined by means of flameless atomic absorption spectrophotometry. Zinc was present in a concentration about 100 p.p.m. (dry weight), copper in a concentration about 5 p.p.m., and lead in slightly more than 1 p.p.m. Analysis of various levels along the cranio-caudal axis of the rat spinal cord revealed differences in the heavy metal content. The Timm sulfide silver staining method has demonstrated that metals in the spinal cord have a distinct regional distribution. To obtain a differentiation between the stainable metals, the effects of six chelating agents (DEDTC, dithizone, oxine, EDTA, dipyridyl, and phenantroline) on the Timm pattern were tested. EDTA left the pattern unchanged, while the other compounds showed individual differences in their influence on the Timm pattern, suggesting that the heavy metal pattern of the spinal cord consists of multiple compartments. The effect of intravital multiple low dose treatment with three of the chelating agents on the histochemical pattern and the metal content of the spinal cord was also investigated. It was found that a decrease in the metal content was not followed by reduction of stainability and vice versa.     

The dithizone,Timm's sulphide silver and the selenium methods demonstrate a chelatable pool of zinc in CNS

Summary From rats intravitally treated with dithizone (diphenyl-thiocarbazone) brains and spinal cords were removed and freeze-dried. The dithizonates present in the CNS tissue were extracted with carbon tetrachloride and subjected to a multielement analysis (proton activation, PIXE). It was found that the extract contained two metals. Most of the metal was zinc, but small traces of copper were also dectected. Because prior treatment with the chelating agent, dithizone, can block both the Timm and the selenium metal staining methods, it is suggested that the three techniques label predominantly zine in the neuropil (DTS-zine).     

The dithizone, Timm's sulphide silver and the selenium methods demonstrate a chelatable pool of zinc in CNS. A proton activation (PIXE) analysis of carbon tetrachloride extracts from rat brains and spinal cords intravitally treated with dithizone

From rats intravitally treated with dithizone (diphenyl-thiocarbazone) brains and spinal cords were removed and freeze-dried. The dithizonates present in the CNS tissue were extracted with carbon tetrachloride and subjected to a multielement analysis (proton activation, PIXE). It was found that the extract contained two metals. Most of the metal was zinc, but small traces of copper were also detected. Because prior treatment with the chelating agent, dithizone, can block both the Timm and the selenium metal staining methods, it is suggested that the three techniques label predominantly zinc in the neuropil (DTS-zinc).     

Partitioning of zinc and copper within subnuclear nucleoprotein particles.

Nuclei from frozen calf thymus suspended in buffer were analyzed for metal content prior to and after repeated washing. After three such extractions about 0.1 micrograms Zn/mg DNA and 0.025 micrograms Cu/mg DNA remained tightly associated with chromatin. This level of metal was essentially unchanged with subsequent washings. Digestion of extracted nuclei with micrococcal nuclease yielded soluble nucleoprotein containing zinc and copper. Metal enriched regions of chromatin appeared to be preferentially solubilized by digestion, and the solubilized metal was only partially dializable either with or without EDTA. Metal profiles generated from gel (A-5m) chromatography analysis of chelated and non-chelated solubilized chromatin were distinctive in that copper was undetectable (by flame AA) while zinc was associated only with low molecular weight products when EDTA was used. In contrast, both metals were detected with higher molecular weight oligonucleosomes in the absence of chelating agents. Additionally, the two metals localized within nucleoprotein peaks and these metal-containing regions were only resolved by gel chromatography when EDTA was omitted throughout the procedure. A discrete Cu-rich species in a region of the profile suggests a subset of Cu-rich nucleoprotein complexes.     

EDTA chelation effects on urinary losses of cadmium, calcium, chromium, cobalt, copper, lead, magnesium, and zinc

The efficacy of a chelating agent in binding a given metal in a biological system depends on the binding constants of the chelator for the particular metals in the system, the concentration of the metals, and the presence and concentrations of other ligands competing for the metals in question. In this study, we make a comparison of the in vitro binding constants for the chelator, ethylenediaminetetraacetic acid, with the quantitative urinary excretion of the metals measured before and after EDTA infusion in 16 patients. There were significant increases in lead, zinc, cadmium, and calcium, and these increases roughly corresponded to the expected relative increases predicted by the EDTA-metal-binding constants as measured in vitro. There were no significant increases in urinary cobalt, chromium, or copper as a result of EDTA infusion. The actual increase in cobalt could be entirely attributed to the cobalt content of the cyanocobalamin that was added to the infusion. Although copper did increase in the post-EDTA specimens, the increase was not statistically significant. In the case of magnesium, there was a net retention of approximately 85% following chelation. These data demonstrate that EDTA chelation therapy results in significantly increased urinary losses of lead, zinc, cadmium, and calcium following EDTA chelation therapy. There were no significant changes in cobalt, chromium, or copper and a retention of magnesium. These effects are likely to have significant effects on nutrient concentrations and interactions and partially explain the clinical improvements seen in patients undergoing EDTA chelation therapy.     

Extraction,Recovery, and Biostability of EDTA for Remediation of Heavy Metal-Contaminated Soil

Chelation removal of heavy metals from contaminated soil is seen as a viable remediation technique. A useful chelating agent should be strong, reusable, and biostable during metal extraction and recovery operations. This work tested the extraction, recovery, and biostability of EDTA as a potential remediating agent. Parameters, including EDTA concentration, soil type, soil content, washing cycle, precipitant concentration and type, and pH, were varied and tested during metal extraction and recovery operations. Factors, including EDTA concentration, aqueous and 5% soil slurry, presence of Pb, acclimated and unacclimated activated sludges, along with abiotic control, were varied and studied in the biodegradation of EDTA. The results showed that EDTA was able to extract lead completely from the tested soils, amenable to recovery by addition of cationic and anionic precipitants in the alkaline pH range, relatively biostable even under conditions very favorable toward biodegradation. Thus, EDTA is a strong, recoverable, and relatively biostable chelating agent that has potential for soil remediation application.     

Remediation of Metal-Contaminated Soil by an Integrated Soil Washing-Electrolysis Process

Chelating agents such as EDTA and DTPA are often used to remove metals from soil. However, their toxicity, bio-recalcitrance, and problems with recovery of heavy metal and chelating agents severely limit their applications. A biodegradable chelating agent, LED3A, and two surfactants, SDS and Triton X 100, were evaluated as potential alternatives for remediation of metal-contaminated soil.

LED3A alone only removed 40% of cadmium the addition of surfactant significantly enhanced its cadmium removal capacity up to 80% for a wide range of pH (5 to 11). The enhancement increased with both surfactant concentrations and LED3A concentrations. Because LED3A had a much higher removal capacity for copper, the synergistic effect of surfactant-LED3A mixture was less obvious. Sequential extraction analysis indicated that the LED3A not only removed copper from carbonate and Fe-Mn oxide fraction, but also from organic fractions. A three-dimension electrolysis reactor could effectively recover both metals and LED3A-SDS within thirty minutes. The combined soil washing by LED3A-surfactants and electrolysis provides a potential approach for remediation of copper- and cadmium-contaminated soils.     

几种有机添加剂对遏蓝菜和东南景天吸收提取Zn的效应

通过盆栽试验,比较研究乙二胺四乙酸二钠盐(EDTA)、味精废液、柠檬酸、乙酸、草酸和混合试剂(柠檬酸∶味精废液∶EDTA∶KC l=10∶1∶2∶3)对Zn超累积植物遏蓝菜(T h lasp i caeru lescens)和东南景天(S edum a lf red ii)吸收提取Zn的影响。结果表明:各种添加剂均提高土壤中的水提取态和NH4NO3提取态Zn的含量,其顺序为EDTA混合试剂>味精废液>有机酸。除乙酸和味精废液外,其余添加剂都显著促进遏蓝菜的生长,以混合试剂的增产效果最好;但只有EDTA和混合试剂在浓度为10mm o l/kg土时提高了东南景天的生物量。混合试剂在浓度为6~10 mm o l/kg土时促进遏蓝菜对Zn的吸收和向地上部转移;EDTA和浓度为10mm o l/kg的混合试剂能显著促进东南景天对Zn的吸收和向地上部的转移。因此东南景天配合环境风险较小、用量为10mm o l/kg土的混合试剂较适合我国南方的Zn污染土壤。     

Effect of EDTA and Tannic Acid on the Removal of Cd,Ni, Pb and Cu from Artificially Contaminated Soil by Althaea rosea Cavan

In this study an ornamental plant of Althaea rosea Cavan was investigated for its potential use in the removal of Cd, Ni, Pb and Cu from an artificially contaminated soil. Effect of two different chelating agents on the removal has also been studied by using EDTA (ethylenediaminetetracetic acid) and TA (tannic acid). Both EDTA and TA have led to higher heavy metal concentration in shoots and leaves compared to control plants. However EDTA is generally known as an effective agent in metal solubilisation of soil, in this study, TA was found more effective to induce metal accumulation in Althaea rosea Cavan under the studied conditions. In addition to this, EDTA is toxic to some species and restraining the growth of the plants. The higher BCF (Bio Concentration Factor) and TF (Translocation Factor) values obtained from stems and leaves by the effects of the chemical enhancers (EDTA and TA) show that Althaea rosea Cavan is a hyper accumulator for the studied metals and may be cultivated to clean the contaminated soils.     

Simultaneous Desorption and Desorption Kinetics of Phenanthrene,Anthracene, and Heavy Metals from Kaolinite with Different Organic Matter Content

Soils contaminated simultaneously with polycyclic aromatic hydrocarbons (PAHs) and heavy metals pose major threat to human health and environment by getting released from soil into water environment. The purpose of this study was to evaluate simultaneous desorption and desorption kinetics of PAHs (phenanthrene and anthracene) and heavy metals (lead, nickel, and zinc) from artificially contaminated kaolinite soils with different organic matter content. Batch desorption tests were conducted using single and combined enhancing agents containing Triton X-100 and Tween 80 as non-ionic surfactants, Ethylenediaminetetraacetic acid (EDTA) as a chelating agent, and citric acid as an organic acid. The solution with the highest removal efficiency was the combined solution of Triton X-100 (10% w/w) + EDTA (0.01 M). Removal levels around 92, 46, 92, 95, and 96% were obtained for phenanthrene, anthracene, lead, nickel, and zinc, respectively, by using this combination. Batch desorption kinetics experiments were performed using the mentioned combination. During the first 24 h, desorption kinetics were rapid, followed by a plateau until the end. The data obtained from desorption kinetics experiments were fitted with four kinetics models: pseudo-second-order equation, empirical power function, elovich, and parabolic diffusion. The correlation coefficient of the pseudo-second-order equation was higher than that of other functions. Moreover, batch experiments have showed inverse correlations between removal efficiency and organic matter content of soil.     

The effects of chelating agents on radical generation in alkaline peroxide systems, and the relevance to substrate damage

A spin-trapping EPR technique has been employed to explore the generation of hydroxyl radicals from reactions between a series of first row transition metal ions and aqueous hydrogen peroxide at pH 10, and with a range of chelating agents (EDTA, DTPMP and the readily biodegradable ligands S,S-EDDS and IDS). In the absence of these chelating agents only Cu(II) generates a significant level of hydroxyl radicals; in their presence with Cu(II) EDTA and IDS give similar behaviour whereas EDDS and DTPMP inhibit hydroxyl radical generation. For Fe(II), EDTA, DTPMP and IDS significantly enhance ^√OH production under these conditions whereas EDDS does not. Results from model cellulose damage experiments broadly confirm the findings for copper, though experiments with Fe(II) lead to somewhat contrasting results. Our findings are discussed in terms of binding constants and implications for alkaline peroxygen bleaching systems.     

Effects of EDTA and EDDS on Heavy Metal Activation and Accumulation of Metals by Soybean in Alkaline Soil

The application of chelating agents for phytoextraction has demonstrated that it is an efficient method to activate heavy metals in polluted soil. We conducted pot experiments using soybean, which has been considered an indicator plant, to study the effects of EDTA and EDDS on heavy metals’ activation, and on the soybean. The study results indicated that EDDS decreased the chlorophyll content of the leaves and increased the malondialdehyde (MDA) content of the soybean. EDTA also decreased the chlorophyll content of the leaves. EDDS had a strong influence on activating Cu (2583-8900-fold) and Zn. The addition of 5 mmol kg^?1 of EDDS markedly increased the uptake of metals. Compared with the control, EDDS increased the Cu uptake (100-205-fold). EDTA greatly increased the activation of heavy metals; it also increased Cu uptake in a concentration-dependent manner. EDTA also increased the biological concentration factor (BCF) and the transfer factor (TF) in a concentration-dependent manner. The BCF and the TF reached maximum levels when 5 mmol kg^?1 EDDS was applied to the pots.     

Application of in situ cage cultures of phytoplankton for monitoring heavy metal pollution in two Norwegian fjords

A simple apparatus for in situ use of the cage culture technique for growing planktonic algae has been developed and used to follow growth and metal uptake of three diatoms in two Norwegian fjords polluted by heavy metals. Continuous pumping of sea water through a chelating resin and continuous water sampling was used to obtain average values for dissolved and particulate heavy metal content at the various test sites. The metals investigated (zinc, copper, lead, cadmium, and mercury) showed differences in the proportion of dissolved to particulate fractions.The three species of diatoms tested gave systematic growth responses to heavy metal pollution in the moderately polluted fjord; one alga died, one showed reduced growth rate at the more contaminated site compared with those at less polluted sites, while the most tolerant alga was apparently not affected. In the heavily polluted fjord only the most tolerant alga survived, showing decreasing growth rate with increasing pollution.Uptake of heavy metals increased generally with increasing heavy metal content in the sea water. The contents in the algae grown in the most polluted fjord were much higher than those obtained in the less polluted fjord, which were higher than the contents reported for algae from non-contaminated areas.     

Influence of Complexation on the Uptake by Plants of Iron, Manganese, Copper and Zinc: II. EFFECT OF DTPA IN A MULTI-METAL AND COMPUTER SIMULATION STUDY

Barley (Hordeum vulgare L.) plants were grown in nutrient solutionscontaining the chelating agent, DTPA. The experiments replicatedthose reported in the preceding paper in which EDTA was thechelating agent used. The concentrations of all the chemicalspecies of metals were stimulated using the program NUTRIENT.The concentrations of DTPA used were chosen to give a similarrange of complexation as used in the EDTA experiments. The effectof complexation by DTPA on the uptakes of the metal ions Fe³⁺,Mn²⁺, Cu²⁺, and Zn²⁺ and on plant growth were sufficiently differentfrom those with EDTA to indicate some dependence on the natureof the chelating agent used. The biggest difference betweenthe EDTA and DTPA experiments occurred in the solutions containingthe largest concentrations of these reagents. With DTPA, chlorosiswas seen only in the early stages; otherwise the plants showednormal growth. A simple chemical model for metal uptake is discussed. Key words: DTPA, EDTA, micronutrients, trace metals, computer simulation, plants, absorption, iron, manganese, copper, zinc     

Detection and Analysis of 12 Heavy Metals in Blood and Hair Sample from a General Population of Pearl River Delta Area

To detect the content of 12 heavy metals in blood and hair sample from a general population of Pearl River Delta area, and to analyze the influence of duration of residence, gender, age, smoking and drinking on the heavy metal content. Use inductively coupled plasma mass spectrometry to detect the content of 12 heavy metals lead (Pb), mercury (Hg), cadmium (Cd), aluminum (Al), arsenic (As), copper (Cu), chrome (Cr), manganese (Mn), nickel (Ni), zinc (Zn), tin (Sn) and antimony (Sb) in blood and hair samples of a total of 50 subjects from a general population, collected by multistage stratified cluster random sampling method. The geometric mean of heavy metal content in blood samples of general population (μg/L): blood aluminum 214.00; blood chrome 92.82; blood manganese 21.43; blood nickel 20.59; blood copper 0.67; blood zinc 11.50; blood arsenic 0.55; blood cadmium 2.45; blood tin 0.00; blood antimony 1.92; blood lead 158.84; and blood mercury 1.19. The geometric mean of heavy metal content in hair samples of general population (μg/g): hair aluminum is 84.65; hair chrome 0.00; hair manganese 2.44; hair nickel 0.61; hair copper 28.49; hair zinc 136.65; hair arsenic 0.75; hair cadmium 0.46; hair tin 1.04; hair antimony 0.05; hair lead 8.97; and hair mercury 0.69. Some heavy metals were correlated with duration of residence, gender, age, smoking and drinking. This was the first time that simultaneously detecting heavy metal content in blood and hair was used to analyze the internal heavy metal burden in resident population of Pearl River Delta area. These data can serve as reference for further research.     

Sorption of Heavy Metals to the Filamentous Bacterium Thiothrix Strain A1

A study was undertaken to determine the ability of the filamentous bacterium Thiothrix strain A1 to sorb heavy metals from solution. Cells of Thiothrix strain A1 were harvested, washed, and suspended in solutions of metals. After an equilibration period, biomass was separated from solution and the metal content in acid-digested cells and/or filtrates was determined by atomic absorption spectrophotometry. Sorption of nickel and zinc was very rapid; most of the sorbed metal was bound in less than 10 min. The sorption data for copper fit the Freundlich isotherm, and nickel and zinc data fit biphasic Freundlich isotherms. Sorption of both nickel and zinc was dependent on cell age. Cells harvested 24 h after inoculation sorbed approximately one-half of the amount of metal per gram cell protein than did cells harvested after 48, 72, or 96 h. Calcium and magnesium effectively competed with zinc for binding sites, whereas potassium had only a slight effect on the capacity of cells to sorb zinc. The primary mechanism of metal sorption apparently was ion exchange, because 66 to 75% of nickel or zinc could be desorbed by placing metal-laden cells in a solution of 5 mM CaCl₂. A competition experiment with nickel and zinc indicated that both metals occupied the same sorption sites. The strong chelating agents EDTA and NTA effectively prevented metal uptake, but lactate enhanced the uptake of nickel. Thiothrix strain A1 grown in nickel-containing medium had a relatively low uptake of nickel compared with uptake by resting cells suspended in a simple buffer solution.     

Concentrations of heavy metals in maternal and umbilical cord blood

Concentrations of lead, cadmium, methylmercury and total mercury were measured in maternal and umbilical cord blood using graphite atomic absorption spectrometry. Two essential metals, copper and zinc, were also determined using ion chromatography. Lead, copper and zinc were found to be lower in the cord blood, whereas methylmercury and total mercury were higher in cord blood than in maternal blood. Little differences were noted for cadmium in maternal and cord blood. Significant positive correlations were observed between the concentrations in maternal and cord blood with regard to lead (correlation coefficient, r = 0.44), copper (r = 0.34), zinc (r = 0.29), methylmercury (r = 0.44) and total mercury (r = 0.58). These results suggest that, like essential metals, most heavy metals can move rather freely across the human placenta. The potential health effects of heavy metal transfer from mothers to young infants cannot be discounted.     

Assessment of Some Heavy Metals in the Dead Sea Mud and Treatment Optimization

This paper describes the experimental remediation of the Dead Sea mud and the quantitative determination of some heavy metals. Herein, two chelating agents were employed as extracting aqueous solution: ethylenediaminetetraacetic acid (EDTA) and citric acid. The study focused on the main known heavy metals that were reported previously to be in the Dead Sea mud, which are Co, Ni, Pb, Zn, and Cr. Findings had indicated that citric acid was efficient in the removal of the aforementioned heavy metals. Physicochemical parameters that were expected to affect the removal of metals in the Dead Sea mud were optimized. Those parameters were the chelating agent concentration, mixing time and speed, type of washing water, temperature, and pH. The results showed that the best removal of heavy metals from Dead Sea mud can be achieved under optimum citric acid concentration, 1.5 g/50 mL for treatment of 10 g mud. Optimum mixing speed and time were found to be 800 rpm and 1 hr, respectively. Regarding washing water, it was found that the use of the same water for repeated washing provided better removal percentages. pH values and temperature had effect on removal percentages of the heavy metals from mud. However, working at pH 7 and room temperature would provide convenient results for heavy metal removal.     

Bioaccumulation of metal cations by Saccharomyces cerevisiae

Yeast cells are capable of accumulation of various heavy metals, preferentially accumulating those of potential toxicity and also those of value. They retain their ability to accumulate heavy metals under a wide range of ambient conditions. In the present study it was shown that yeast cells in suspension accumulate heavy metal cations such as Cu²⁺, Co²⁺. The level of copper accumulation was dependent on the ambient metal concentration and was markedly inhibited by extremes of ambient pH. Temperature (5–40°C) and the presence of the alkali metal sodium had much smaller effects on the level of copper accumulation. This suggests that in waste-waters of pH 5.0–9.0, yeast biomass could provide an effective bioaccumlator for removal and/or recovery of the metal. During bioaccumulation and subsequent processes it is necessary to retain the biomass. It was shown in the present study that this could be achieved by cell immobilization. Immobilization allowed for complete removal of Cu²⁺, Co²⁺, and Cd²⁺ from synthetic metal solutions. The immobilized material could be freed of metals by use of the chelating agent ethylenediamine tetraacetic acid (EDTA) and recycled for further bioaccumulation events with little loss of accumulation capacity.Correspondence to: J. R. Duncan     

Effect of Chelating Agents and Metal Ions on Gametangial Formation in the Moss Bryum argenteum Hedw

Chelating agents such as EDTA and EDDHA markedly stimulate theformation of gametangia in the dioecious moss Bryum argenteum,and the effect is more pronounced on male than on female clones.EDTA-stimulated gametangial formation is associated with significantincreases in endogenous iron and copper. Ferric citrate alsoenhances gametangial formation, but copper sulphate is inhibitory.The present findings are discussed in the context of earlierinvestigations on other plants in an attempt to explain thepossible involvement of chelating agents and metal ions in stimulatingthe onset of the reproductive phase in this moss. Bryum argenteum, gametangial formation, chelating agents, metal ions