Speciation of trace element compounds in samples of biota from marine ecosystems

Abstract

The toxicity, mobility, bioavailability and bioaccumulation of metals are dependent on the particular physico-chemical form in which the element occurs in the environment. Special attention has been paid to metals which are essential for the proper functioning of organisms if present in appropriate amounts but are toxic if in excess (i.e. Se, Cr), and also to non-essential elements (i.e. Hg, Pb, Cd, Sn and As). To assess the potential hazard to the health of marine organisms, qualitative and quantitative analyses of metal species accumulating along the food chain needs to be carried out. This paper reviews the available information on the speciation of trace elements in the food chain in marine ecosystems and the analytical tools used for acquiring reliable information in this field. Advantages and limitations of commonly used techniques indicate that all metal species in different samples need diverse extraction, separation and detection conditions. Although not recommending which procedure is the most suitable to determine a given compound, speciation analysis has the potential to be a powerful tool for the identification of trace element species in biological samples.     

Role of nuclear analytical probe techniques in biological trace-element research

Many biomedical experiments require the qualitative and quantitative localization of trace elements with high sensitivity and good spatial resolution. The feasibility of measuring the chemical form of the elements, the time course of trace element metabolism, and conducting experiments in living biological systems are also important requirements for biological trace element research. Nuclear analytical techniques that employ ion or photon beams have grown in importance in the past decade and have led to several new experimental approaches. Some of the important features of these methods are reviewed here along with their role in trace element research. Examples of their use are given to illustrate potential for new research directions. It is emphasized that the effective application of these methods necessitates a closely integrated multidisciplinary scientific team.     

Laboratory gloves as a source of trace element contamination

Contamination in a trace element laboratory can come from a variety of sources, including laboratory gloves. Therefore, vinyl and latex gloves were obtained from as many manufacturers as would supply gloves. These gloves were either prepared for acid-washing and subsequent soaking in an acid solution, or immersed in an acid solution for a duration of either 1 min or 1 h. Incubation washes were analyzed for a variety of trace elements by flame atomic abosrption spectroscopy (AAS) or inductively coupled mass spectrometry (ICP-MS). Results indicated that only three brands of vinyl gloves were acceptable for use in a trace element laboratory, whereas others had contamination of different elements. Latex gloves contained such high levels of biologically important elements that they were not considered suitable for routine trace element work. Vinyl gloves of choice should be routinely acid-washed before use in a trace element laboratory.     

Imaging and speciation of trace elements in biological environment

Mineral elements, often at the trace level, play a considerable role in physiology and pathology of biological systems. Metallogenomics, metalloproteomics, and metallomics are among the emerging disciplines which are critically dependent on spatially resolved concentration maps of trace elements in a cell or tissue, on information on chemical speciation, and on that on metal-binding coordination sites. The mini-review discusses recent progress in analytical techniques for element profiling on the genome scale, biological trace element imaging, and probing, identification and quantification of chemical species in the biological environment. Imaging of the element distribution in cells and tissue sections is becoming possible with sub-micrometer spatial resolution and picogram-level sensitivity owing to advances in laser ablation MS, ion beam and synchrotron radiation X-ray fluorescence microprobes. Progress in nanoflow chromatography and capillary electrophoresis coupled with element specific ICP MS and molecule-specific electrospray MS/MS and MALDI enables speciation of elements in microsamples in a complex biological environment. Laser ablation ICP MS, micro-SXRF, and micro-PIXE allow mapping of trace element distribution in 1D and 2D proteomics gels. The increasing sensitivity of EXAFS and XANES owing to the use of more intense synchrotron beams and efficient focusing optics provide information about oxidation state, fingerprint speciation of metal sites and metal-site structures.     

Quantitative imaging of metals in tissues

Metals and other trace elements play an important role in many physiological processes in all biological systems. Characterization of precise metal concentrations, their spatial distribution, and chemical speciation in individual cells and cell compartments will provide much needed information to explore the metallome in health and disease. Synchrotron-based X-ray fluorescent microscopy (SXRF) is the ideal tool to quantitatively measure trace elements with high sensitivity at high resolution. SXRF is based on the intrinsic fluorescent properties of each element and is therefore element specific. Recent advances in synchrotron technology and optimization of sample preparation have made it possible to image metals in mammalian tissue with submicron resolution. In combination with correlative methods, SXRF can now, for example, determine the amount and oxidation state of trace elements in intra-cellular compartments and identify cell-specific changes in the metal ion content during development or disease progression.     

同步辐射X射线吸收近边结构光谱技术在磷素固相形态研究中的应用

固相形态磷是控制环境中磷素生物可利用性、迁移流失能力的重要形态.基于同步辐射光源的X射线吸收近边结构(XANES)光谱技术可在分子水平上识别目标元素周围的局部化学信息,在非破坏性、原位直接表征等方面体现出其独特的优越性,成为表征化学物质存在形态和阐明化学反应微观机制的前沿技术之一,在环境化学领域中得到了广泛关注.本文简述了磷的XANES的基本理论,综述了XANES技术在矿物、土壤及有机肥中磷素固相形态研究中的应用进展,并分析了该技术应用在环境介质中磷形态表征中所面临的挑战及发展趋势,指出XANES技术应与其他微观光谱技术及宏观试验方法有机结合,多种表征技术取长补短,以期为环境介质中磷素形态表征及转化机制研究提供全面有效的技术支撑.     

Tissues and organs as indicators of intestinal absorption of minerals and trace elements,evaluated in rats

Tissue and organ deposition and blood parameters were evaluated as indices of mineral and trace element absorption in rats. The absorption of elements was quantified in relation to nitrogen retention, i.e., considering the weight gain and new tissue synthesis. A rapeseed meal diet was supplied with three levels of calcium, two levels of zinc, and two levels of copper in a factorial design. In general, an increase in dietary mineral content increased the relative absorption, which in turn, increased the tissue deposition progressively. Striated muscle, however, did not respond to either an increased calcium or zinc supply. Furthermore, an increased calcium absorption caused a depression of the fractional phosphorus and magnesium content of femur bones. The copper content of the kidneys and the heart muscle was directly proportional to the amount of absorbed zinc and iron, respectively. The iron content of tissues was, in general, inversely proportional to zinc absorption and showed a tendency to be directly proportional to copper absorption. The zinc level in tissues was, in a similar way, inversely correlated to measured calcium absorption. In conclusion, interactions between elements do not only affect the intestinal element absorption, but also the distribution of already absorbed elements in tissues and organs.     

Dephytinization of a rat diet

Soaking of a rat diet, high in both plant phytate and phytase, progressively degraded the phytate content with time of soaking. This dephytinization in turn enhanced the digestion of feed organic matter in the animals, and it significantly improved the absorption and retention of minerals and trace elements as observed in balance studies. Incorporation of elements into specific tissues was evaluated as a reflection of bioavailability. Some tissues did reflect the preceding absorption of certain elements; other tissues seemed less suitable as indicators of trace element absorption. Dietary calcium addition in many ways contrasted the soaking procedure: inorganic calcium addition to the feed reduced phosphorus, magnesium, and trace element bioavailability, and interfered with the internal deposition of the elements. The external dephytinization of the feed did not affect the phosphohydrolase activity of the intestinal mucosa as manifested by alkaline phosphatase activity or phytase activity. The mucosal phytase and alkaline phosphatase activities were, however, mutually correlated, supporting the view that “phytase” activity is a less substratespecific action of alkaline phosphatase activity or a fraction of this activity.     

Studies on some trace and minor elements in blood

Blood is one of the widely used specimens for biological trace element research because of its biological significance and ease of sampling. We have conducted a study of the blood of the Kalpakkam township population for trace and minor elements. For this purpose, analytical methods have been developed and standardized in our laboratory for the elemental analysis of blood plasma and red cells. Inductively coupled plasma-mass spectrometry (ICP-MS), a relatively new technique, has been applied for the analysis of trace elements. Details regarding spectral interference and matrix interference encountered in the analysis of blood and the methods of correcting them have been discussed. Flame atomic absorption spectrometry (AAS)/atomic emission spectrometry (AES) has been applied for the determination of minor elements. Precision and accuracy of these methods have also been discussed.     

Microbial Manganese Oxidation in the Lower Mississippi River: Methods and Evidence

Abstract

Recent work has led to the suggestion that biologically-mediated redox processes might be important in the regulation of dissolved trace element concentrations in rivers, especially with regard to manganese. Here, we focus on the removal of dissolved Mn from lower Mississippi River water. Experiments indicate that dissolved Mn can be rapidly removed from lower Mississippi River water on a timescale of days or less and that Mn oxides are formed. However, demonstrating a biological origin for this removal is problematic. Experiments reveal that commonly used microbial controls, including NaN₃ mixtures, HgCl₂, heat sterilization, and sonification all affect fluvial particulate Mn through dissolution, disaggregation, interference with adsorption, or particle ageing. Thus, these microbial controls may affect abiotic as well as biological processes. Evidence supporting microbial removal of dissolved Mn from lower Mississippi River water includes a temperature optimum for the process (～30°C), a lower activation energy than reported for heterogeneous inorganic Mn oxidation, and a faster rate than reported for autocatalytic inorganic Mn oxidation. This rapid Mn oxidation process occurs at essentially the same rate in the dark as well as the light. Observation of Mn removal at similar rates in a blackwater river in addition to the lower Mississippi, suggests that this is a common phenomenon in fluvial systems. If, as has been shown by other lab studies, the freshly biologically precipitated Mn oxides have a high specific surface area, then our observations provide a potential link between microbial activity, Mn cycling, and the cycling of other particle-reactive trace elements in rivers. Our results also indicate that unfiltered river water samples for dissolved Mn analysis should be filtered as soon as possible or at least stored cold if immediate filtration is not possible.     

不同利用强度下绿洲农田土壤微量元素有效含量特征

以地处极端干旱背景下的塔里木盆地南缘策勒绿洲为研究区,以绿洲化为视角,根据当地农民耕作习惯于2005年选择4块试验农田,分别代表当地典型的农田利用强度。基于单项指数(Ei)和综合指数(Ec)相结合的评价方法,分析了绿洲农田在不同利用强度下的土壤Fe、Cu、Mn、Zn等微量元素有效性,探讨了土壤微量元素与人为耕作管理措施之间的关系。结果表明:人为耕作管理强度会对土壤微量元素有效含量产生重要影响,绿洲不同位置农田因施肥强度和管理方式的差别,其土壤微量元素有效含量特征存在显著差异。绿洲内部农田土壤有效Fe、Cu、Mn、Zn含量显著高于绿洲边缘各样地,新垦农田土壤有效Fe、Cu、Zn含量均低于对照样地;绿洲内部农田土壤微量元素有效性最高,新垦农田最低;农田土壤微量元素有效性与土壤有机质存在显著的正相关关系。     

Concentrations of copper, zinc, manganese, rubidium, and magnesium in thoracic empyemata and corresponding sera

In this study, a number of selected trace elements and clinically relevant parameters were compared between thoracic empyemata and the corresponding sera for a better understanding of the trace element distribution between these two compartments. Serum-empyema pairs were obtained from 13 patients and quantified for selected and essential trace elements, namely copper (Cu), zinc (Zn), manganese (Mn), rubidium (Rb), and magnesium (Mg), by inductively coupled plasma-mass spectrometry (ICP-MS). In addition, the concentrations of the following clinical laboratory parameters were analyzed by standard methods: total protein, leukocyte count, lactate dehydrogenase, glucose, pH, and the C-reactive protein. Individual concentrations of the elements determined in the empyemata were frequently higher than in pleural effusions of any other benign or malignant condition except for Cu. Serum Cu exceeded the normal range (600–1400 μg/kg) in 6 out of 13 patients (median 1410 μg/kg). In the empyemata, Zn concentrations (median 2000 μg/kg) were characteristically higher than in the sera (median 450 μg/kg) and exceeded the upper limit for serum (1200 μg/kg) in 8 of the 13 patients. Manganese concentrations in the empyemata (median 2.7 μg/kg) were also higher compared to corresponding sera, although they stayed within the limits considered normal for serum of healthy adults (upper limit 2.9 μg/kg). Rubidium was also moderately higher in most empyemata (median 290 μg/kg) and exceeded the upper limit for serum (560 μg/kg) in two patients. The median concentration of the essential element magnesium was higher in the empyemata (23 mg/kg) than in the sera (21 mg/kg). However, all serum Mg concentrations except three remained within the normal range (17–22 mg/kg). Removal of large amounts of empyematous fluid may deprive the body of trace elements and can cause suboptimal or deficient trace element status and homeostasis. Recuperation will be accelerated by compensatory supplementation of trace elements. Therefore, selective medication with adequate trace element compounds in patients with thoracic empyema can be generally recommended for zinc. The other elements need not necessarily be monitored or substituted, because of their stable concentrations in the serum. Rb may have a biological impact, but deficiency symptoms in man are not clearly defined. Deceased.     

Chemical Speciation and Mobility of Some Trace Elements in Vermicomposted Fly Ash

Fly ash, a by-product of power plants, is currently being used extensively in India for soil amendment. However, the toxic elements sorbed in the fly ash might pose a serious threat to the environment, causing soil and water contamination. Vermicomposting of fly ash is expected to reduce the contamination of toxic trace metal and could improve the mobility of essential trace element. The current study is focused on characterizing different species of trace metals and their bio-availability in the vermicomposted fly ash (VCFA)-treated lateritic soil. As a fertilizer, different doses (10%, 20%, 30%, 40%, and 50%) of VCFA were applied to the soil and sequential extraction was carried out to analyze trace elements. In the different fractions, Cr < Mn < Pb < Fe were found to be sorbed more to Fe-Mn oxide-bound fractions, whereas Cd, Cu, and Zn were bound more to organic-matter-bound fractions; Cr and Ni were mostly bound to residual fraction. The Fe-Mn oxides and organic-matter-bound fractions may be bio-available with the appropriate environmental condition, whereas chromium and nickel mostly associated with residual fraction are very difficult to release into the environment. The mobility factor index showed the midlevel substitution (i.e., 10% to 30% of VCFA to lateritic soil) to be beneficial as these doses increased the bio-availability of some essential trace elements and restricted the availability toxic trace metals in the soil. At higher doses, the toxic trace metals were found to be released in the bio-available form, which could be hazardous to the environment.     

Concentations of copper, Zinc, and magnesium in sera from patients with idiopathic dilated cardiomyopathy

Trace elements are known to have a key role in myocardial metabolism. The accumulation (cobalt, arsenic, copper) or deficiency (selenium, zinc) of trace elements may be responsible for idiopathic dilated cardiomyopathy. We investigated the trace element concentrations (Cu, Zn, Mg) in sera from patients with dilated cardiomyopathy by atomic absorption spectrophotometry. We observed that patients with dilated cardiomyopathies have higher copper and lower zinc concentrations in serum than healthy controls. The magnesium concentrations of patients did not differ significantly from that of control subjects.     

Trace elements in the human central nervous system studied with neutron activation analysis

The central nervous system (CNS) should be especially sensitive to disturbances in trace element concentrations because of its high metabolic rate and low capacity for regeneration. Comparatively few studies have been made on trace elements in the CNS, which prompted us to begin a study of trace elements in four different brain lobes of the CNS, as well as in the spinal cord. Samples were obtained at autopsy and handled carefully in order to avoid contamination. They were freeze-dried and sealed in quartz tubes that were irradiated in a nuclear reactor. A simple chemical separation into six fractions was performed. The gamma spectra for these fractions was registered using a Ge(Li) detector and a computerized multichannel analyzer. Results for the following elements were obtained: Ca, Cd, Co, Cr, Cs, Cu, Fe, Rb, Se, and Zn, as well as for Na and K (not reported). Other elements were also detected in some samples. Using this technique, brain samples from ten patients with Alzheimer’s disease and ten control cases were examined.     

发网菌目黏菌子实体微量元素的初步分析

运用能散X-射线微区分析技术研究了发网菌目黏菌代表种的子实体的微量元素组成,结果表明:分属于7属的7种供试发网黏菌子实体囊被上的微量元素组成存在差异。尽管在显微镜下观察不到可见的如绒泡菌目黏菌子实体中存在的石灰质颗粒,但其中的钙元素所占比例均仍较高。同时,元素组成上的差异与子实体的颜色没有相关性。     

Investigation of the role and chemical form of iron in the ovarian carcinogenesis process

BackgroundOvarian cancer is one of the most frequent types of gynaecological malignancy among women. Despite the advances in diagnostic techniques, ovarian tumours are still detected at a late stage, thus the survival rate is very low. Iron is an essential metal in the human body, yet its potential role in ovarian carcinogenesis is yet to be determined. The aim of this study was to check if iron oxidation state in tissue and cystic fluid can be treated as an indicator of the malignancy of the ovarian tumours. Another aspect of this study was to investigate the role of iron in carcinogenesis mechanism in ovarian tumour transformation.MethodsSynchrotron radiation X-ray absorption near edge structure (SR-XANES) spectroscopy was used to analyze the human ovarian tumour tissues and cystic fluids of different types and grades of malignancy. Fresh, non-fixed, frozen samples were used to analyze the state of iron oxidation in all the biological materials. The samples were obtained from patients requiring surgical intervention. The High Energy X-ray Absorption Spectroscopy (XANES) measurements were performed at the beamline P65 at Petra III Extension, DESY – Deutsches Elektronen - Synchrotron.ResultsFe XANES spectra were collected at selected points of a few different regions of the samples. For each specimen, an average of these points was probed. Having been measured, the spectra were compared with organic and inorganic reference materials. Also, the position of the absorption edge was calculated using the integration method. In all specimens, iron occurred in the oxidation states, Fe2+ and Fe3+, although the fraction of iron in the third oxidation state was substantial, especially in malignant cases. The results also show differences in the chemical form of iron in the tissue and cystic fluids of the same patient.ConclusionsThe cryo-XANES measurement carried out for ovarian cancer tissues and cystic fluids showed changes in the chemical form of iron between non-malignant and malignant tumours. For both types of sample can be observed that they contain iron on second and iron on third oxidation state. Moreover, the tendency was observed that malignant tumours of the ovary contain a larger fraction of iron in the second oxidation state compared to non-malignant ones.     

Trace element profiles in cancer patients

It has become evident over the last two decades that there is an intimate relationship between the trace elements and cancer. Some trace elements have been shown to be carcinogens, others appear to provide protection against cancer. Profound changes in trace element concentrations and distribution occur in patients with cancer, but most changes remain undefined. A review of a number of studies of trace element changes in patients with cancer demonstrates that simple correlations of trace element levels in disease are of only limited use. Such reports underscore the need for large-scale studies that consider the many variables of malignancies and of trace element chemistry. The variables that must be considered for cancer include tissue of origin; histologic, pathologic and clinical staging; nutritional status as reflected by serum levels of calcium, iron, magnesium, phosphorus, the electrolytes, pH, albumen, and globulin; endocrine balance, effects of previous and concurrent therapies such as surgery, chemotherapy, hormonal manipulation, immunotherapy, and radiotherapy; history of exposure to toxic agents; and the presence of other disease. Similarly, trace element studies entail variables that must be considered and controlled prospectively, including timing and techniques of sampling, storage, and analysis, and simultaneous measurement of at least the majority of possibly interrelated elements rather than studying one element at a time. The various national cooperative oncology groups such as ECOG, SWOG, and SEOG now offer unusually well-studied groups of cancer patients who are managed according to carefully and prospectively defined protocols in participating institutions. With present knowledge, it is now time to approach these groups with a proposal to incorporate trace element studies in their protocols. A potential protocol will be discussed.     

蒙药材多叶棘豆中五种微量元素的形态分析

本文采用水煎法对蒙药材多叶棘豆中微量元素进行提取,并分别用微孔滤膜分离技术、大孔吸附树脂柱分离法、萃取法等分离方法对水煎液中元素各形态进行分离。利用FAAS法测定了原药材以及水煎液各形态中Cu、Zn、Mn、Fe和Ca五种微量元素含量并进行了形态分析。分析结果表明,水煎液中五种元素的总提取率在41.7%～87.0%,残留率在9.1%～26.4%,可溶态百分率在16.6%～80.0%,可溶态中五种元素的有机态与无机态的比例为36.9%～250%;该测定方法对各元素的加标回收率在90.3%～113.7%。多叶棘豆水煎液可溶态中Zn、Mn、Fe、Ca元素是以亲水性有机结合形态为主,Cu元素以无机离子型为主,多种形态共存的复杂体系。     

中国生物微量元素研究的现状与展望

微量元素包括铁、锌、铜、锰、硒、碘等,在机体生命活动中发挥着极其重要的生理学功能,参与蛋白质结构,并通过与蛋白质和其他有机基团结合参与酶、激素和维生素等生物大分子合成。微量元素代谢失衡会发生系列病理改变并最终导致疾病。生物微量元素研究是多学科交叉的科学研究领域,业已成为生命科学的国际研究热点。该领域发展对人类生存与健康具有重要意义。经过几代科学家的不懈努力,我国的生物微量元素研究已经从无到有,逐步取得令国际同行瞩目的科研成果,尤其近年来,我国该领域基础研究快速发展,更是相继产生了诸多高水平原创性科研成果。这种迅速发展与壮大,得益于大批海外归国优秀人才,更离不开国家和社会在科研经费和政策等方面的大力支持。简介了近年来我国生物微量元素研究领域的科学家及其主要科研成果,期望广大读者能全面了解国内生物微量元素研究现状以及可喜的发展趋势。衷心期待着中国的生物微量元素研究能有更大发展,能有更大成就。