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Four molecular forms of transferrins with different iron-binding states were separated by HPLC using a pyridinium polymer column. The elution order was monoferric transferrin bound to the C-site, holotransferrin, apotransferrin and monoferric transferrin bound to the N-site. Human sera were also analyzed with the column, and ICP-MS combined with HPLC was used to detect iron in each peak. Transferrin peaks separated by HPLC were also confirmed by an immunological method. The percentages of iron saturation in transferrins obtained by the HPLC method were compared with the values calculated from clinical data.
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PMID:Separation of human serum transferrins with different iron-binding states by high-performance liquid chromatography using a pyridinium polymer column. 1186 94

The feathers of two Ardeid species, the Little Egret (Egretta garzetta) and the Black-crowned Night Heron (Nycticorax nycticorax) were collected from six egretries and two egretries respectively, located in different areas in the New Territories of Hong Kong, including the Mai Po Marshes (within a Ramsar site). These feathers were digested and concentrations (microg/g dry weight) of copper (4.6-19.4), iron (8.1-641.3), manganese (0.4-19.4), zinc (51.3-183.5), lead (0.1-5.1), cadmium (0.01-0.15), chromium (0.06-1.7) and mercury (0.0-7.1) were determined by ICP-AES, ICP-MS and CVAAS. The levels of manganese, mercury and lead found were equal to or less than the concentrations found in previous investigations, reflecting a slight downward trend most apparent with lead. As a general rule, the levels of lead and mercury were higher in the egretries close to the polluted Deep Bay. A probabilistic risk assessment of the possible adverse effects on the breeding success of the Little Egret was carried out with respect to mercury, lead and cadmium. It was concluded that mercury (0.5-7.1 microg/g dry weight feathers) probably has had adverse effects at the Au Tau egretry of the Little Egrets, but there was no evidence of adverse effects at other egretries. The probabilistic analysis also indicated a low likelihood of adverse effects of mercury on the breeding of the Black-crowned Night Herons at A Chau (0.3-1.2 microg/g) and Mai Po Village (0.0-1.4 microg/g). The evidence for the effects of lead and cadmium was limited but suggested there may possibly be adverse effects with lead but not cadmium.
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PMID:Risk to breeding success of Ardeids by contaminants in Hong Kong: evidence from trace metals in feathers. 1189

The gene encoding for the methionyl aminopeptidase from the hyperthermophilic archaeon Pyrococcus furiosus (PfMetAP-II; EC 3.4.11.18) has been inserted into a pET 27b(+) vector and overexpressed in Escherichia coli. The new expression system resulted in a 5-fold increase in purified enzyme obtained from a 5 L fermentor growth. The as-purified PfMetAP-II enzyme, to which no exogenous metal ions or EDTA was added, was found to have 1.2 equiv of zinc and 0.1 equiv of iron present by ICP-AES analysis. This enzyme had a specific activity of 5 units/mg, a 60-fold decrease from the fully loaded Fe(II) enzymes. When an additional 2 equiv of Zn(II) was added to the as-purified PfMetAP-II, no activity could be detected. The combination of these data with previously reported whole cell studies on EcMetAP-I further supports the suggestion that the in vivo metal ion for all MetAP's is Fe(II). Both Co(II)- and Fe(II)-loaded PfMetAP-II showed similar substrate specificities to EcMetAP-I. Substrate binding was largely affected by the amino acid in the P1 position and the length of the polypeptide. The substrates MSSHRWDW and MP-p-NA showed the smallest K(m) values while the substrates MGMM and MP-p-NA provided the highest turnover. The catalytic efficiency (k(cat)/K(m)) of PfMetAP-II for MP-p-NA at 30 degrees C was 799 500 and 340 930 M(-1) s(-1) for Co(II)- and Fe(II)-loaded PfMetAP-II, respectively. Maximum catalytic activity was obtained with 1 equiv of Co(II) or Fe(II), and the dissociation constants (K(d)) for the first metal binding site were found to be 50 +/- 15 and 20 +/- 15 nM for Co(II)- and Fe(II)-substituted PfMetAP-II, respectively. Electronic absorption spectral titration of a 1 mM sample of apo-PfMetAP-II with Co(II) provided a dissociation constant of 0.35 +/- 0.02 mM for the second metal binding site, a 17500-fold increase compared to the first metal binding site. The electronic absorption data also indicated that both Co(II) ions reside in a pentacoordinate geometry. PfMetAP-II shows unique thermostability and the optimal temperature for substrate turnover was found to be approximately 85 degrees C at pH 7.5 in 25 mM Hepes and 150 mM KCl buffer. The hydrolysis of MGMM was measured in triplicate between 25 and 85 degrees C at eight substrate concentrations ranging from 2 to 20 mM. Both specific activity and K(m) values increased with increasing temperature. An Arrhenius plot was constructed from the k(cat) values and was found to be linear over the temperature range 25-85 degrees C, indicating that the rate-limiting step in PfMetAP-II peptide hydrolysis does not change as a function of temperature. Co(II)- and Fe(II)-loaded PfMetAP-II have similar activation energies (13.3 and 19.4 kJ/mol, respectively). The thermodynamic parameters calculated at 25 degrees C are as follows: DeltaG++ = 46.23 kJ/mol, DeltaH++ = 10.79 kJ/mol, and DeltaS++ = -119.72 J.mol(-1).K(-1) for Co(II)-loaded PfMetAP; DeltaG++ = 46.44 kJ/mol, DeltaH++ = 16.94 kJ/mol, and DeltaS++ = -99.67 J.mol(-1).K(-1) for Fe(II)-loaded PfMetAP. Interestingly, at higher temperatures (> 50 degrees C), Fe(II)-loaded PfMetAP-II is more active (1.4-fold at 85 degrees C) than Co(II)-loaded PfMetAP-II.
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PMID:Overexpression and divalent metal binding properties of the methionyl aminopeptidase from Pyrococcus furiosus. 1204 50

An electrothermal atomic absorption (ETAAS) method for the determination of traces of iron (0.1-1.0 microgram g-1) in Fe-doped indium phosphide (InP) has been developed. In order to overcome the indium matrix-effect and to achieve a useful detection limit, a preliminary solvent-extraction of Fe(III) with acetylacetone (HAA) is necessary. After sample dissolution with hydrochloric acid (1 + 1) the digest is evaporated to dryness, Fe(II) is oxidized to Fe(III) with nitric acid, the residue is dissolved in 0.01 mol L-1 HCl and the iron is extracted at pH 2.0 with 0.5 mol L-1 HAA in toluene. The organic phase is injected into the graphite furnace and the iron is directly evaluated by external organic standard calibration. The limit of detection (3SB) resulting from further in-situ preconcentration is 0.03 microgram g-1. When the method was applied to the analysis of real samples containing 0.2-0.7 microgram g-1 Fe, the RSD was in the range 8-21%. Results were compared with those independently obtained on the decomposed sample solution with inductively coupled atomic emission spectrometry (ICP-AES). The detection limit of the ICP-AES method, that needs matrix-matched standards, is 0.20 microgram g-1.
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PMID:Determination of traces of iron in indium phosphide by electrothermal atomic absorption spectrometry combined with solvent extraction. 1212 54

Mining and processing of lead (Pb)/zinc (Zn) ore at the Anvil Range mine occurred near the town of Faro in the Yukon Territory, Canada, for approximately 30 years, beginning in 1968. A study was undertaken to examine whether the mining activities had left a detectable "footprint" on the environment in the way of heavy metal phytoaccumulation. Foliage of three native plant species was sampled: bog blueberry (Vaccinium uliginosum), Labrador tea (Ledum groenlandicum), and willow (Salix sp.), at approximately 0.25, 2.5, 12, 30, and 200 (control) km distant from the mill (ore-processing facility at the mine). Foliage samples were oven-dried, wet- or dry-ashed, and analyzed for metal content using ICP-AES. In addition to Pb and Zn, the primary ore constituents, copper (Cu), iron (Fe), and cadmium (Cd), were also assayed. As expected, foliar Pb and Zn concentrations were elevated in plants at the sites closest to the mill, i.e., 0.25 and 2.5 km from the mine facility. Copper and Fe, both essential nutrients for plants, were also elevated in foliage at the sites closest to the mill, but not to a level that would be of concern. Foliar Cd levels were highest in Salix relative to the other species but were not affected by proximity to the mill. Results suggest that Ledum may be the best indicator of high environmental concentrations of Pb, while Salix may be the best indicator of elevated Zn and Cd.
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PMID:Heavy metal (Pb, Zn, Cd, Fe, and Cu) contents of plant foliage near the Anvil Range lead/zinc mine, Faro, Yukon Territory. 1229 90

Tin is considered to be a priority contaminant by the Codex Alimentarius Commission. Tin can enter foods either from natural sources, environmental pollution, packaging material or pesticides. Higher concentrations are found in processed food and canned foods. Dissolution of the tinplate depends on the of food matrix, acidity, presence of oxidising reagents (anthocyanin, nitrate, iron and copper) presence of air (oxygen) in the headspace, time and storage temperature. To reduce corrosion and dissolution of tin, nowadays cans are usually lacquered, which gives a marked reduction of tin migration into the food product. Due to the lack of modern validated published methods for food products, an ICP-AES (Inductively coupled plasma-atomic emission spectroscopy) method has been developed and evaluated. This technique is available in many laboratories in the food industry and is more sensitive than atomic absorption. Conditions of sample preparation and spectroscopic parameters for tin measurement by axial ICP-AES were investigated for their ruggedness. Two methods of preparation involving high-pressure ashing or microwave digestion in volumetric flasks were evaluated. They gave complete recovery of tin with similar accuracy and precision. Recoveries of tin from spiked products with two levels of tin were in the range 99+/-5%. Robust relative repeatabilities and intermediate reproducibilities were <5% for different food matrices containing >30 mg/kg of tin. Internal standard correction (indium or strontium) did not improve the method performance. Three emission lines for tin were tested (189.927, 283.998 and 235.485 nm) but only 189.927 nm was found to be robust enough with respect to interferences, especially at low tin concentrations. The LOQ (limit of quantification) was around 0.8 mg/kg at 189.927 nm. A survey of tin content in a range of canned foods is given.
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PMID:Determination of total tin in canned food using inductively coupled plasma atomic emission spectroscopy. 1232 43

The speciation of trace elements in serum samples of hemodialysed patients was investigated using on-line connection of SEC and ICP-MS. The 0.02 mol/l TRIS-HCl buffer of pH 7.5 was used as mobile phase. The results of speciation as well as the total concentration data were compared with those of control group of healthy person. Alterations of total concentration were observed in case of selenium and zinc only. Iron was present in form of transferrin and ferritin, main amount of copper was bound to ceruloplasmin and selenium compounds were identified as selenoproteine P and glutathione peroxidase. The latter compound was detected in samples of control group only. The chromatograms of the other elements were similar and no substantial changes between both investigated groups were observed.
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PMID:Speciation of Cu, Se, Zn and Fe in blood serum of hemodialysed patients. 1244 34

Iron overload augments diseases of the liver and microorganism infection as well as deregulates the immune system. In vitro analysis of the effects of iron loading and its chelation involves determining the amount of iron constituting overload, which metal sources and cell lines to use and reliable assay methods. The uptake of 500 microM FeSO4 or FeEDTA by CEMss, U937 or leukocytes was confirmed by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). Excess iron increased CEMss viability (assessed by MTT, XTT, Trypan- and Alamar Blue) by an average of 18% (P = 0.034). Flow cytometry indicated dye-viable cells to be undergoing apotosis/necrosis while still confirming an increase (9%, P < 0.001) in excess iron-induced viability. The iron chelator desferioxamine (DFO) when added in addition to Fe, reversed the effects of excess iron (and vice versa) and had detrimental effects when used on its own (33% inhibition of viability as measured by dyes and 10.85%; P = 0.0427 assessed by flow cytometry). The 4 dyes demonstrated different levels of sensitivity in detecting the influence of iron or DFO but gave a related, qualitative picture while flow cytometry and ICP-AES data was more quantitative.
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PMID:Comparing qualitative and quantitative spectroscopic techniques for the detection of the effect of direct iron loading of mammalian cell cultures. 1248 27

An estimation of the dietary exposure of French consumers to 21 essential and non-essential mineral elements using duplicate meals (breakfast and lunch) purchased from catering establishments was investigated after digestion by a closed vessel microwave procedure and quantification by ICP-MS. Daily dietary exposure estimates for metals and minerals were compared with the Provisional Tolerable Weekly Intakes (PTWI), the Tolerable Daily Intake (TDI) or the Acceptable Daily Intakes (ADI), as established by the FAO/WHO to estimate the risk of toxicity, and the US Recommended Daily Allowances (US RDA) or the Estimate Safe & Adequate Daily Dietary Intakes (ESADDI). Moreover, comparisons were made with those from previous French studies as well as those from other countries. The estimated mean daily intakes were 11 microgram for lithium, 3.42 g for sodium, 192 mg for magnesium, 2.03 mg for aluminium, 3.64 g for potassium, 642 mg for calcium, 154 microgram for chromium, 12.3 mg for iron, 2.15 mg for manganese, 4 microgram for cobalt, 74 microgram for nickel, 925 microgram for copper, 10.2 mg for zinc, 147 microgram for arsenic, 66 microgram for selenium, 112 microgram for molybdenum, 3.6 microgram for cadmium, 2.32 mg for tin, 3 microgram for antimony, 9 microgram for mercury and 34 microgram for lead. For the non-essential (toxic) elements, aluminium, tin, antimony, cadmium, arsenic, mercury and lead, the daily intake estimates were far below tolerable limits; and similar or somewhat lower than their respective PTWI, ADI, TDI, ESADDI and US RDA for individual minerals and essential trace elements, with good agreement with other country studies. The performance of the multi-elemental ICP-MS technique was also evaluated.
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PMID:Determination of several elements in duplicate meals from catering establishments using closed vessel microwave digestion with inductively coupled plasma mass spectrometry detection: estimation of daily dietary intake. 1251 18

Seven household treatment technologies for the removal of arsenic (Alcan, BUET, DPHE/DANIDA, Garnet, Sono, Stevens, Tetrahedron) were each evaluated using water from 63 different tube wells taken from 3 different regions of Bangladesh. The technologies that were evaluated were chosen from those that appeared user friendly, readily available and whose promoters were open to participate in the study. Arsenic concentrations in feed and treated waters were analysed by the PeCo 75 arsenic field test kit, AA-hydride generation and ICP-AES. Feed water arsenic concentrations were found to be up to 600 microg l(-1). The more advanced treatment methods using: activated alumina (Alcan, BUET); metallic iron (Sono); anionic exchange resin (Tetrahedron) and iron coagulation (Stevens) were found to be most easily used and efficiently reduced arsenic concentrations to below the Bangladesh drinking water standard (0.05 mg As l(-1)). The use of aluminium sulphate coagulants and permanganate oxidants in the DPHE/DANIDA technology introduced unacceptably high concentrations of aluminium and manganese into the treated waters and are not recommended in household water treatment applications. While arseric concentrations were initially considered to be of paramount importance, it became clear that such technologies can increase the risk of bacterial contamination in the treated water and this needs serious consideration as this could create a hazard much greater than the arsenic contained in the water. Ground waters sampled during the course of this study were mostly found to be bacteria free. To minimize any risks relating to bacterial contamination the addition of hypochlorite or the boiling of water is necessary.
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PMID:A field based evaluation of household arsenic removal technologies for the treatment of drinking water. 1252 10


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