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Speciation analysis of arsenic and selenium compounds by capillary electrophoresis.

High-performance capillary electrophoresis is applied to the separation of different inorganic and organic arsenic and selenium compounds. In comparison with UV-detection, an approach with conductivity detection is described expecting higher sensitivity and universality. In this case the capillary was statically modified with CTAB before the electromigration procedure. The separation was performed with an electrolyte system consisting of CHES and Triton X-100. Detection limits of 0.06 mg/L or lower were obtained for As(V) and Se(VI). Water samples of an arsenic-polluted tailing of tin mining processes were analysed for anions as well as arsenic and selenium species.

Journal Article↗

Speciation of metals and metalloids in sediments with LTGC/ICP-MS.

Low temperature GC on-line coupled with an ICP/MS has been used as analytical device to detect organometallic and -metalloid compounds from various sediments of rivers and harbours in West Germany. The different species have been volatilized by derivatization with NaBH(4) and cryofocussed in a trap. Different alkylated species of As, Sn and Sb have been identified, while only methylated compounds of the elements Ge, Se, Te, Hg and Bi could be detected.

Journal Article↗

A high performance liquid chromatography - inductively coupled plasma-mass spectrometry interface employing desolvation for speciation studies of platinum in chemotherapy drugs.

A novel interface between high performance liquid chromatography (HPLC) and inductively coupled plasma-mass spectrometry (ICP-MS) is described. The eluent from the HPLC is nebulised into a heated cyclone spray-chamber and the solvent removed using a Nafion membrane drier, held at 75 degrees C, and a cryogenic condenser. The condenser consists of 6 Peltier heat pumps connected to liquid cooled aluminium blocks. At a nebuliser gas flow rate of 0.6 l min(-1), the membrane drier removes 58% of the vapour and the Peltier condenser 75% of the remaining vapour, i.e. a total desolvation efficiency of 89%. This enables the use of HPLC solvents which otherwise would destabilise the ICP, e.g. 100% acetonitrile or methanol, and permits the use of solvent gradients with minimal baseline drift. The system has been applied to the determination of platinum species in an organoplatinum drug used for chemotherapy in human plasma ultrafiltrate of patients treated with this new drug (JM-216). The limit of detection for platinum species has been 0.6 ng nl(-1) (i.e. 120 pg of Pt) and several species have been separated with good resolution.

Journal Article↗

High-performance flow atomic spectrometry: New nebulization techniques, on-line speciation and on-line sample pretreatment.

An interface-free combination of HPLC separation techniques and methods for element determination by atomic spectrometry can be achieved by hydraulic high-pressure nebulization (HHPN). With high-temperature HHPN (300 ( degrees )C) super heated liquids can be nebulized providing aerosol yields of up to 90% in flame AAS. This new nebulization method combines the advantages of HHPN and thermospray techniques (very small aerosol droplets, high aerosol yield, nebulization of saturated salt solutions).

Journal Article↗

Speciation of chromium by coupled column HPLC-ICP-MS - the effects of interfering ions.

ICP-MS is used as an element selective detector for chromium species in aquatic samples using a coupled columm system consisting of a cation guard column and an anion column. The effects of large concentrations of disturbing ions, like NO(3)(-), Cl(-), CO(3)(2-) and SO(4)(2-), are discussed and the comparison with the use of different chromium isotopes for data acquisition is given. Quite low concentrations such as 100 mg/l of CO(3)(2-), 10 mg/l Cl(-) and even 5 mg/l SO(4)(2-) can cause an overestimation of chromium if not properly resolved1995 / Accepted: 24 October 1995

Journal Article↗

Speciation of organolead compounds in water samples by GC-AAS after in situ butylation with tetrabutylammonium tetrabutylborate.

A simple and reliable analytical method for the determination of organolead compounds in water samples has been developed by means of a gas chromatographic-atomic absorption system (GC-AAS). The derivatization of the organolead cations, necessary before their determination, was directly achieved in the water samples by the reagent tetrabutylammonium tetrabutylborate. Hexane/pentane was used to extract the butylated tetraorganolead compounds. The organic layer was dried and purified with sodium sulfate, concentrated by evaporation and injected into a GC-AAS system. The effects of various parameters on the derivatization and extraction procedures were studied and discussed.

Journal Article↗

Simultaneous determination of tetrathiomolybdates and molybdates in spiked blood plasma by differential pulse voltammetry and their speciation.

A selective, sensitive and reliable voltam- metric method for the simultaneous determination of MoS(2-)(4) and MoO(2-)(4) has been developed. The reduction of HgMoS(4) at -0.43 V and the reduction of Mo as complex of MoO(2)-oxine(2) at -0.63 V in acetate buffer at pH 4.9 are utilized for the simultaneous determination by differential pulse cathodic stripping voltametry (DPCSV). Cadmium and lead form complexes with MoS(2-)(4), but do not interfere with the determination. The limit of detection is 0.59 microg/l Mo for MoS(2-)(4) and 1.8 microg/l Mo for MoO(2-)(4). The R.S.D at a concentration level of 20 microg/l is 4.7% for MoO(2-)(4) and 3.6% for MoS(2-)(4). The method is applied to spiked blood plasma samples for the determination of free tetrathiomolybdates and molybdates. Additionally, labile tetrathiomolybdates and molybdates can be determined in spiked blood plasma after treatment of the sample with 0.15 mol/l KCN and ultrafiltration.

Journal Article↗

Speciation in Chlamydia: genomewide phylogenetic analyses identified a reliable set of acquired genes.

Horizontal gene transfer (HGT), a process through which genomes acquire sequences from distantly related organisms, is believed to be a major source of genetic diversity in bacteria. A central question concerning the impact of HGT on bacterial genome evolution is the proportion of horizontally transferred sequences within genomes. This issue, however, remains unresolved because the various methods developed to detect potential HGT events identify different sets of genes. The present-day consensus is that phylogenetic analysis of individual genes is still the most objective and accurate approach for determining the occurrence and directionality of HGT. Here we present a genome-scale phylogenetic analysis of protein-encoding genes from five closely related Chlamydia, identifying a reliable set of sequences that have arisen via HGT since the divergence of the Chlamydia lineage. According to our knowledge, this is the first systematic phylogenetic inference-based attempt to establish a reliable set of acquired genes in a bacterial genome. Although Chlamydia are obligate intracellular parasites of higher eukaryotes, and thus suspected to be isolated from HGT more than the free-living species, our results show that their diversification has involved the introduction of foreign sequences into their genome. Furthermore, we also identified a complete set of genes that have undergone deletion, duplication, or rearrangement during this evolutionary period leading to the radiation of Chlamydia species. Our analysis may provide a deeper insight into how these medically important pathogens emerged and evolved from a common ancestor.

Base Composition↗

Hair mercury speciation as a function of gender, age, and body mass index in inhabitants of the Negro River basin, Amazon, Brazil.

Human mercury contamination can be monitored through hair analysis of mercury's inorganic and organic form as methylmercury (Me-Hg). Hair total mercury and Me-Hg were studied in a Negro River fish-eating population in relation to age, gender, and body mass index (BMI). This riverbank population eats fish at least twice a day and is exposed to high levels of Me-Hg. Total mercury ranged from 1.51 microg/g to 59.01 microg/g, with only 21% of the sampled population having Hg concentration of less than 10 microg/g hair. The mean percentage of Me-Hg was 71.3% (range 34% to 100%) of the total mercury in hair. No statistically significant differences were found in regard to age groups (children and adults) or BMI. However, women had significantly lower total mercury in hair than men, but the percentage of Me-Hg was not significantly different. Women in fertile age (15-40 years) had hair total mercury ranging from 1.65-32.63 microg/g, and 65% in this subgroup had hair mercury above 10 microg/g hair. The percentage of Me-Hg concentration in hair of this freshwater, fish-eating population is comparable to populations eating ocean fish from different parts of the world and does not seem to be affected by age, gender, and BMI.

Adolescent↗

A multidisciplinary approach to assess history, environmental risks, and remediation feasability of soils contaminated by metallurgical activities. Part B: direct metal speciation in the solid phase.

Three soils contaminated by industrial smelting activities previously characterized using chemical and physical approaches (Part A, Venditti et al. [2000] Arch Environ Contam Toxicol [in press]) were further investigated through a complementary mineralogical approach. X-ray diffraction allowed identification of the main mineral components: carbonates (calcite, siderite) and iron oxides (magnetite, hematite, goethite) prevailed in both siderurgical soils, and siliceous compounds were more abundant in the agricultural soil. Scanning electron microscopy coupled with energy dispersive x-ray microanalysis showed that carbonates and oxides contained low amounts of heavy metals (HMs), but the potential solubility of such species under acidic conditions associated to their abundance may result in a high mobility of associated contaminants. Microanalysis also revealed peculiar minor mineral forms (metallic nodules, alloys, ore residues) that were highly informative about the history of soil contamination. Small metal-rich particles were observed (brass, lead and zinc oxides). In such compounds, HMs showed high chemical stability and were consequently poorly mobile and toxic, but also poorly removable through chemical leaching. A risk of mechanical dispersion was suggested for these particles, particularly after the chemical and/or mechanical disruption of carbonate cements. These features allowed completion and confirmation of the conclusions achieved after chemical and physical investigations, providing accurate information about the history, environmental risks and remediation feasability of industrial soil contamination.

Feasibility Studies↗

Axenic culture of reptilian Blastocystis isolates in monophasic medium and speciation by karyotypic typing.

The growth of axenic reptilian isolates of Blastocystis in Iscove's modified Dulbecco's medium (IMDM) was studied and the morphology of the parasite was examined by phase-contrast microscopy. The chromosomal patterns of these reptilian isolates of Blastocystis were examined by pulsed-field gel electrophoresis (PFGE) and compared with those of B. hominis and B. lapemi, a sea snake Blastocystis. IMDM with 10% horse serum supported excellent growth of the reptilian Blastocystis isolates. The parasites from all the isolates were predominantly vacuolar, but multivacuolar and amoeboid forms were also seen. Amoeboid forms with rather elongate pseudopodia were also observed. There were some differences in size, morphology, and growth characteristics in the different reptilian isolates. The karyotypic patterns of the Blastocystis isolates from tortoise, iguana, and python were distinctly different from one another and from those obtained with B. hominis and B. lapemi. On the basis of the above-mentioned differences in chromosomal patterns, the tortoise, iguana, and python isolates are described as new species, viz., B. geocheloni sp. nov. from Geochelone carbonaria (red-footed tortoise), B. cycluri sp. nov. from Cyclura cornuta (rhino iguana), and B. pythoni sp. nov. from Python reticulatus (reticulated python).

Animals↗

Speciation and structure of ferriprotoporphyrin IX in aqueous solution: spectroscopic and diffusion measurements demonstrate dimerization, but not mu-oxo dimer formation.

Changes in epsilon (393) (the Soret band) of aqueous ferriprotoporphyrin IX [Fe(III)PPIX] with concentration indicate that it dimerizes, but does not form higher aggregates. Diffusion measurements support this observation. The diffusion coefficient of aqueous Fe(III)PPIX is half that of the hydrated monomeric dicyano complex. Much of the apparent instability of aqueous Fe(III)PPIX solutions could be attributed to adsorption onto glass and plastic surfaces. However, epsilon (347) was found to be independent of the aggregation state of the porphyrin and was used to correct for the effects of adsorption. The UV-vis spectrum of the aqueous dimer is not consistent with that expected for a mu-oxo dimer and the (1)H NMR spectrum is characteristic of five-coordinate, high-spin Fe(III)PPIX. Magnetic susceptibility measurements using the Evans method showed that there is no antiferromagnetic coupling in the dimer. By contrast, when the mu-oxo dimer is induced in 10% aqueous pyridine, characteristic UV-vis and (1)H NMR spectra of this species are observed and the magnetic moment is consistent with strong antiferromagnetic coupling. We propose a model in which the spontaneously formed aqueous Fe(III)PPIX dimer involves noncovalent interaction of the unligated faces of two five-coordinate H(2)O/HO-Fe(III)PPIX molecules, with the axial H(2)O/OH(-) ligands directed outwards. This arrangement is consistent with the crystal structures of related five-coordinate iron(III) porphyrins and accounts for the observed pH dependence of the dimerization constant and the spectra of the monomer and dimer. Structures for the aqueous dimer are proposed on the basis of molecular dynamics/simulated annealing calculations using a force field previously developed for modeling metalloporphyrins.

Adsorption↗

Speciation of trace elements in proteins in human and bovine serum by size exclusion chromatography and inductively coupled plasma-mass spectrometry with a magnetic sector mass spectrometer.

Proteins are separated by size exclusion chromatography while atomic ions from the inorganic elements are detected on-line by inductively coupled plasma-mass spectrometry. A double focusing mass analyzer provides very high sensitivity, low background, and sufficient spectral resolution to separate the atomic ions of interest from most polyatomic ions at the same nominal m/z value. The chromatograms show the distribution of the elements of interest between protein-bound and free fractions and provide the approximate molecular weights of those protein fractions that contain the elements monitored. The distribution of various elements, including V, Mo, Fe, Co, Mn, and lanthanides, in human or bovine serum samples are shown. Alkali metals and Tl are present primarily as free metal ions and are not bound to proteins. Inorganic elements spiked into the serum samples can be followed into various proteins. EDTA does not remove Fe, Pb, Sn, or Th from the proteins but does extract Mn from some proteins. Procedures for determining the effects of breaking disulfide linkages on the metal binding characteristics of proteins are also described.

Animals↗

Speciation of termite gut protists: the role of bacterial symbionts.

At least 12 termite gut protists have been named because of their bacterial symbionts. Dozens more species are diagnosed by epi- and endosymbionts and more still have regular bacterial associations referred to in their species description. Molecular systematic studies have begun to identify these bacteria, but the ecological relations with their protist bionts are still unknown. Recent findings of acetogenic spirochetes in termite guts may explain the peculiar arrangement of spirochetes on some of these protists. Other bacteria function as motility or chemotactic symbionts of these protists. The size and shape of the parabasal body, a Golgi complex, are morphological characters of the Parabasalia (trichomonads, hypermastigids) that may be influenced by regular, heritable epi- and endosymbiotic bacteria.

Animals↗

Effects of flooding on lead and cadmium speciation in sediments from a drinking water reservoir.

Rímov water reservoir on the river Malse is the main source of drinking water for the town of Ceské Budejovice and for the majority of inhabitants in the South Bohemian region, Czech Republic. Changes in cadmium and lead contents in bottom sediments before and after an extensive flood on the river Malse in August, 2002 were therefore determined. A five-step sequential extraction procedure was used in order to obtain more detailed information about the influence of the flood on heavy metal retention. In order to determine the mobility of lead and cadmium, the mobility factor (MF) for these heavy metals was calculated. The mobility factor of cadmium showed a significant decrease in the upper parts of the sediment profiles after the flood (e.g., from 59.4% to 49.1%) caused by a release of cadmium especially from the exchangeable fraction. There were no significant changes in the lead mobility factor after the flood, but a decrease of lead concentration in the exchangeable fraction was observed. Presented results show that the flood led to a leaching of the heavy metals present in bottom sediments into the environment.

Cadmium↗

Molecular speciation of fish sperm phospholipids: large amounts of dipolyunsaturated phosphatidylserine.

The molecular species compositions of the main diacyl phosphoglyceride classes and ether-linked subclasses from sperm of three species of fish, sea bass Dicentrarchus labrax, Atlantic salmon Salmo salar and Chinook salmon Onchorhynchus tsawytscha, were determined. The phospholipids from sperm were highly unsaturated, dipolyunsaturated fatty acid (diPUFA) molecular species comprised 64.6 to 71.8% of phosphatidylserine (PS), 10.1 to 17.4% of phosphatidylethanolamine (PE), and 3.3 to 10.1% of phosphatidylcholine (PC). In sea bass sperm, di22:6n-3 phospholipid was the predominant diPUFA molecular species, but in both salmon species 22:5n-3/22:6n-3 was also a major constituent of PS. Phospholipids containing 22:6n-3 dominated in sea bass sperm with 16:0/22:6n-3 as a major component of PC and PE, and 18:0/22:6n-3 of PE and PS in addition to di22:6n-3 in the latter two classes. In contrast, both salmon species contained much more 20:5n-3 and less 22:6n-3 so that saturated/20:5n-3 and monounsaturated/20:5n-3 molecular species were more abundant than the corresponding molecules containing 22:6n-3. Ether-linked lipids comprised 11.3-36.3% of choline and ethanolamine phosphoglycerides in each fish species. Molecular species containing 22:6n-3 were the major components of 1-O-alkyl-2-acyl-glycerophosphocholine, especially 16:0a/22:6n-3 in sea bass and 18:1a/22:6n-3 in the two salmon species, while in 1-O-alk-1'-enyl-2-acyl-glycerophosphoethanolamine, 16:0a/22:6n-3 was the major component in both salmon and 18:0a/22:6n-3 in sea bass with 18:1a/22:6n-3 abundant in all three species. In Atlantic salmon 1-O-alkyl-2-acyl-glycerophosphoethanolamine comprised 24.6% of ethanolamine glycerophospholipids which were predominantly 16:0a/22:6n-3 and 18:1a/22:6n-3. Phosphatidylinositol from sperm was dominated by stearoyl/C20 PUFA molecular species, in sea bass overwhelmingly 18:0/20:4n-6, while in both salmon species 18:0/20:4n-6 and 18:0/20:5n-3 were equally abundant.

Animals↗

Speciation of tissue and cellular iron with on-line detection by inductively coupled plasma-mass spectrometry.

Iron accumulating to excess in tissues of humans and animal models occurs mainly as complexes with transferrin, ferritin, other hemoproteins, and insoluble hemosiderin particles. To determine the distribution of Fe amongst these molecular species, we have used inductively coupled plasma-mass spectrometry as a means of on-line, isotope-specific detection for their liquid chromatographic separation. The stable isotope 57Fe is a suitable isotope for monitoring the Fe content of each fraction, and its availability at high isotopic enrichment makes it an attractive choice for tracer studies when the use of a radioisotope is undesirable, e.g., in human subjects. The detection system offers the advantages of high sensitivity (detection limits in the parts per billion range), a wide dynamic range (linearity of the calibration curve over several orders of magnitude), and on-line analysis facilitating real-time evaluation of the chromatographic separation, in addition to isotope-specific information. The Fe distributions in healthy rat livers, liver and heart tissue from Fe-loaded human subjects, and human hepatocyte cultures are reported. The ferritin:hemosiderin ratio in these samples is shown to be an indicator of the degree of Fe loading and correlates well with that determined by Zeeman-corrected electrothermal atomic absorption as an alternative means of detection.

Animals↗