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PubMed2005

Cloud point extraction preconcentration prior to high-performance liquid chromatography coupled with cold vapor generation atomic fluorescence spectrometry for speciation analysis of mercury in fish samples.

A cloud point extraction methodology was developed for simultaneous preconcentration of Hg(II), methylmercury (MeHg), ethylmercury (EtHg), and phenylmercury (PhHg) prior to reversed-phase high-performance liquid chromatography (HPLC) on-line coupled with cold vapor atomic fluorescence spectrometry for speciation analysis of mercury in fish. The four mercury species were taken into complexes with ammonium pyrrolidine dithiocarbamate (APDC) in aqueous nonionic surfactant Triton X-114 medium and concentrated in the surfactant-rich phase by bringing the solution to the temperature of 40 degrees C. Baseline separation of the enriched complexes was achieved on an RP-C(18) column with a mixture of methanol, acetonitrile, and water (65:15:20, v/v) containing 200 mmol L(-1) HAc (pH 3.5) as the mobile phase. An on-line postcolumn oxidation of the effluent from HPLC, in the presence of K2S2O8 in HCl, was applied in the system followed by an optimal cold vapor generation of mercury species. The variables affecting the complexation and extraction steps were examined. The preconcentration of 10 mL of solution with 0.08% w/v Triton X-114 and 0.04% w/v APDC at pH 3.5 gave enrichment factors of 29, 43, 80, and 98 for MeHg, EtHg, PhHg, and Hg(II), respectively. Low detection limits (S/N = 3) were obtained, ranging from 2 to 9 ng L(-1) (as Hg) for all species. The developed method was successfully applied to the speciation of mercury in real fish samples.

Animals↗
PubMed2006

Speciation of phytate ion in aqueous solution. Sequestering ability toward mercury(II) cation in NaClaq at different ionic strengths.

As a contribution to understanding the speciation of mercury in the environment and to the study of the sequestering ability of phytate (Phy) toward heavy metal and organometal cations, this paper describes the results of an investigation (at t = 25 degrees C by potentiometry, ISE-H+ glass electrode) of its interactions with mercury(II) cation in NaCl aqueous solutions at different ionic strengths (I = 0.15 and 1.0 mol L(-1)), in the pH range 2.5 < or = pH < or = 9.5 and considering metal-to-ligand ratios of 1:1 < or = Hg/Phy < or = 4:1. The formation of 11 HgiHjPhy(12-2i-j)(- species with i = 1 and 0 < or = j < or = 7 and i = 2 and 0 < or = j < or = 2 was observed. Their complex formation constant values proved to be fairly dependent on ionic strength. The speciation of phytic acid and mercury(II) is also dependent on the metal-to-ligand ratio; the dependence of the stability of phytate-mercury(II) species on the phytate protonation step was modeled, and an empirical predictive relationship was proposed. From the results obtained, phytate has very good sequestering ability toward Hg2+, even in the presence of considerable excesses of chloride ion, that is, another ligand strongly interacting with mercury; this supports future studies both on the use of plants that naturally synthesize it for phytoremediation purposes and on its direct application in remediation techniques.

Cations↗
PubMed2006

Improved arsenic speciation analysis for extracts of commercially available edible marine algae using HPLC-ES-MS/MS.

Their increasing commercial availability and their rather high total arsenic contents necessitate a more detailed arsenic speciation analysis of marine algal products. Compared to current HPLC-ICPMS methods, HPLC on-line with electrospray tandem mass spectrometry in the selected reaction monitoring mode (HPLC-ES-SRM) offers much higher detection selectivity with similarly high sensitivity for most known organoarsenic species. This study demonstrates the advantages of HPLC-ES-SRM for the detection of the main as well as trace arsenic species in extracts of 12 commercially available marine algal powders. The main focus was the unambiguous identification of the detected arsenic species. Four quality control tools were applied for this purpose, including matching chromatographic retention times, comparing SRM transition ratios, recording product ion spectra, and determining accurate masses. As a result, evidence was obtained for the presence of 19 organoarsenic species in the analyzed algal extracts. The method of standard addition was used for quantification. Estimated matrix effects on the analyte signal were similar for most of the investigated arsenic species in extracts of different types of brown algae. This allowed the comparison of the contents of the arsenic species present in the 12 algal extracts on the basis of normalized peak areas. A partial correlation of the arsenic speciation pattern with the algal family or algal order, respectively, was found.

Arsenicals↗
PubMed2003

Beryllium chemical speciation in elemental human biological fluids.

The understanding of beryllium chemistry in human body fluids is important for understanding the prevention and treatment of chronic beryllium disease. Thermodynamic modeling has traditionally been used to study environmental contaminant migration and rarely in the examination of metal (particularly beryllium) toxicology. In this work, a chemical thermodynamic speciation code (MINTEQA2) has been used to model and understand the chemistry of beryllium in simulated human biological fluids such as intracellular, interstitial, and plasma fluids, a number of airway surface fluids for patients with lung conditions, saliva, sweat, urine, bile, gastric juice, and pancreatic fluid. The results show that predicted beryllium solubility and speciation vary markedly between each simulated biological fluid. Formation of beryllium hydroxide and/or phosphate was observed in most of the modeled fluids, and results support the postulation that beryllium absorption in the gastrointestinal tract may be limited by the formation of beryllium phosphate solids. It is also postulated that beryllium is potentially 13% less soluble in the airway surface fluid of a patient with asthma when compared to a "normal" case. The results of this work, supported by experimental validation, can aid in the understanding of beryllium toxicology. Our results can potentially be applied to assessing the feasibility of biological monitoring or chelation treatment of beryllium body burden.

Berylliosis↗
PubMed2004

Arsenic speciation in urine from acute promyelocytic leukemia patients undergoing arsenic trioxide treatment.

Arsenic has been used successfully in clinical trials for treating acute promyelocytic leukemia (APL). Although sublethal doses of inorganic arsenic are used, little is known about the pharmacokinetics and metabolism of the high levels of arsenic in APL patients. To fill this important gap, this study describes the speciation of arsenic in urine from four APL patients treated with arsenic. Each patient was injected daily with an arsenite (As(III)) solution that contained 10 mg of As(2)O(3) precursor. Speciation analysis of the patient urine samples collected consecutively for 48 h, encompassing two intravenous injections of arsenic, revealed the presence of monomethylarsonous acid (MMA(III)), dimethylarsinous acid (DMA(III)), monomethylarsonic acid (MMA(V)), and dimethylarsinic acid (DMA(V)). The intermediate methyl arsenic metabolites, MMA(III) and DMA(III), were detected in most urine samples from all of the patients when a preservative, diethyldithiocarbomate, was added to the urine samples to stabilize these trivalent arsenic species. The major arsenic species detected in the urine samples from the patients were As(III), MMA(V), and DMA(V), accounting for >95% of the total arsenic excreted. The relative proportions of As(III), As(V), MMA(V), and DMA(V) in urine samples collected 24 h after the injections of As(III) were 27.6 +/- 6.1, 2.8 +/- 2.0, 22.8 +/- 8.1, and 43.7 +/- 13.3%, respectively. The relatively lower fraction of the methylated arsenic species in these APL patients under arsenic treatment as compared with that from the general population exposed to much lower levels of arsenic suggests that the high levels of As(III) inhibit the methylation of arsenic (inhibits the formation of methyl arsenic metabolites). The arsenic species excreted into the urine accounted for 32-65% of the total arsenic injected. These results suggest that other pathways of excretion, such as through the bile, may play an important role in eliminating (removing) arsenic from the human body when challenged by high levels of As(III).

Adult↗
PubMed2004

Uranium(VI) solubility and speciation in simulated elemental human biological fluids.

The complete understanding of the human body response to uranium contamination exposure is vital to the development of exposure analysis and subsequent treatments for overexposure. Thermodynamic modeling has traditionally been used to study environmental metal contaminant migration (especially uranium and other radionuclides), allowing examination of chemical processes difficult to study experimentally. However, such techniques are rarely used in the study of metal toxicology. Chemical thermodynamics has a unique and valuable role in developing models to explain metal metabolism and toxicology. Previous computational models of beryllium in simulated biological fluids have been shown to be useful in predicting metal behavior in the human body. However, previous studies utilizing chemical thermodynamics in understanding uranium chemistry in body fluids are limited. Here, a chemical thermodynamic speciation code has been used to model and understand the chemistry of uranium in simulated human biological fluids such as intracellular, interstitial, and plasma fluids, saliva, sweat, urine, bile, gastric juice, pancreatic fluid, and a number of airway surface fluids from patients with acute lung conditions. The results show predicted uranium solubility, and speciation varies markedly between each biological fluid due to differences in fluid composition, ionic strength, and pH. The formation of uranium hydroxide, phosphate (sodium/potassium autunite), and calcium uranate was observed in most of the fluids. The results of this work, supported by experimental validation, can aid in understanding the metabolism and toxic effects of uranium with potential applications to biological monitoring as well as chelation treatment of uranium body burden.

Body Fluids↗
PubMedDate not supplied

Comparison of rac- and meso-2,3-dimercaptosuccinic acids for chelation of mercury and cadmium using chemical speciation models.

The formation constants of various mercury and cadmium chelates of the stereoisomers of 2,3-dimercaptosuccinic acid (DMSA) have been determined from potentiometric titrations in the presence of the competing ligand EDTA. The mercury chelates formed at pH 7.4 are the monomeric HgL of the DMSA diastereoisomers and HHgL2 of rac-DMSA. Mercury is completely complexes at pH greater than 3.0 in solutions containing more than 1 equiv of either rac- or meso-DMSA. At high concentrations (10 microM and above) mercury tends to bind to a greater extent to rac- than to meso-DMSA. At pH 7.4, the predominant cadmium meso-DMSA chelate species in solution is CdL, and HCdL is present at a much smaller concentration. With rac-DMSA, however, the predominant cadmium chelate species is HCdL at a low concentration of the ligand, and at a high concentration of the ligand the species CdL2 predominates. Cadmium is completely chelated at pH 7.4 in solutions containing more than 1 equiv of either rac- or meso-DMSA. At pH around 5.5, which corresponds to the pH of the kidney, however, a significant amount of free cadmium is present in solutions containing 1 equiv or less of either DMSA stereoisomer. From the results of an analysis of speciation models, probable kidney damage, that may result from free cadmium ion release in the kidney during chelation therapy, is inferred when meso-DMSA is used for mobilizing cadmium. In contrast, the release of free cadmium ion is negligible in the pH range in the kidney when rac-DMSA is used. On the basis of the speciation models, rac-DMSA is found to be far superior to meso-DMSA in the treatment of acute cadmium poisoning.

Cadmium↗
PubMed1999

Have chondracanthid copepods co-speciated with their teleost hosts?

Chondracanthid copepods parasites many teleost species and have a mobile larval stage. It has been suggested that copepod parasites, with free-living infective stages that infect hosts by attaching to their external surfaces, will have co-evolved with their hosts. We examined copepods from the genus Chondracanthus and their teleost hosts for evidence of a close co-evolutionary association by comparing host and parasite phylogenies using TreeMap analysis. In general, significant co-speciation was observed and instances of host switching were rare. The prevalence of intra-host speciation events was high relative to other such studies and may relate to the large geographical distances over which hosts are spread.

Animals↗
PubMed2001

Speciation in a ring.

The evolutionary divergence of a single species into two has never been directly observed in nature, primarily because speciation can take a long time to occur. A ring species, in which a chain of intergrading populations encircles a barrier and the terminal forms coexist without interbreeding, provides a situation in which variation in space can be used to infer variation in time. Here we reconstruct the pathway to speciation between two reproductively isolated forms of greenish warbler (Phylloscopus trochiloides). These two taxa do not interbreed in central Siberia but are connected by a long chain of intergrading populations encircling the Tibetan Plateau to the south. Molecular data and climatic history imply that the reproductively isolated taxa came into contact following expansions northward around the western and eastern sides of the plateau. Parallel selection pressures for increased song complexity during the northward expansions have been accompanied by divergence in song structure. Playback experiments show that the two Siberian forms do not recognize each other's songs. Our results show how gradual divergence in a trait involved in mate choice leads to the formation of new species.

Animals↗
PubMed1992

Genetics and speciation.

Called the "mystery of mysteries" by Darwin, speciation is still a little-understood area of evolution. Genetic analysis, however, has yielded new generalizations about speciation and suggests promising avenues of research.

Animals↗
PubMed1994

Sympatric speciation suggested by monophyly of crater lake cichlids.

The existence of sympatric speciation--that populations diverge into species in the absence of physical or ecological barriers--is controversial. The East African Great Lakes harbour hundreds of cichlid species representing only a few monophyletic lineages, although palaeolimnological evidence and local restrictions on species distribution suggest that speciation in these lakes could have been allopatric. The case for sympatry in restricted areas of Lakes Malawi and Tanganyika is stronger but not unassailable. A better case might be made for cichlid species flocks in small, ecologically monotonous crater lakes. Here we present a mitochondrial DNA analysis of cichlid species flocks endemic to two such lakes in Cameroon. The results suggest that the flocks in each lake are monophyletic: the implication being that each lake was colonized once only, the size and shape of each lake being such that subsequent diversification would have been sympatric.

Animals↗
PubMed1995

Speciation driven by natural selection in Drosophila.

Reinforcement is the process by which natural selection strengthens sexual isolation between incipient species, reducing the frequency of maladaptive hybridization and hence completing reproductive isolation. Although this model of speciation was once widely accepted, its plausibility and experimental support have been recently attacked. Here we provide an example of speciation by reinforcement, in the North American fruitfly Drosophila pseudoobscura. The results suggest that females of D. pseudoobscura evolved increased sexual isolation from their sibling species, D. persimilis, by natural selection against maladaptive hybridization.

Animals↗
PubMed1992

A dual level model for speciation by multiple pericentric inversions.

A considerable body of evidence suggests that the deleterious meiotic effects of pericentric inversions in F1 hybrids can be overcome by changes in chiasma location and various means of non-homologous pairing. Such overriding mechanisms may render pericentric inversions benign and increase the likelihood of their fixation in population isolates. It has been argued that overriding mechanisms of this type negate the involvement of pericentric inversions as reproductive isolating mechanisms in speciation. It is suggested, however, that the involvement of pericentric inversions in speciation should be considered on two levels. First, that by reducing meiotic effects in F1 hybrids, overriding mechanisms facilitate the fixation of pericentric inversions. Secondly, when contact hybridization occurs between the chromosomally derived and parental populations second-level effects may be encountered. That is, the recombinational effects of pericentric inversion differences on coadapted gene complexes (sensu Brncic, 1954, Shaw & Coates, 1983) enforce profound inviability barriers in F2 and backcross matings. In this way, multiple pericentric inversions may act as significant post-mating isolating mechanisms, whereas individual inversions with less significant second-level effects may not.

Animals↗
PubMed2003

Engineering evolution to study speciation in yeasts.

The Saccharomyces 'sensu stricto' yeasts are a group of species that will mate with one another, but interspecific pairings produce sterile hybrids. A retrospective analysis of their genomes revealed that translocations between the chromosomes of these species do not correlate with the group's sequence-based phylogeny (that is, translocations do not drive the process of speciation). However, that analysis was unable to infer what contribution such rearrangements make to reproductive isolation between these organisms. Here, we report experiments that take an interventionist, rather than a retrospective approach to studying speciation, by reconfiguring the Saccharomyces cerevisiae genome so that it is collinear with that of Saccharomyces mikatae. We demonstrate that this imposed genomic collinearity allows the generation of interspecific hybrids that produce a large proportion of spores that are viable, but extensively aneuploid. We obtained similar results in crosses between wild-type S. cerevisiae and the naturally collinear species Saccharomyces paradoxus, but not with non-collinear crosses. This controlled comparison of the effect of chromosomal translocation on species barriers suggests a mechanism for the generation of redundancy in the S. cerevisiae genome.

Aneuploidy↗
PubMed2005

Species diversity can drive speciation.

A fundamental question in evolutionary ecology and conservation biology is: why do some areas contain greater species diversity than others? Island biogeographic theory has identified the roles of immigration and extinction in relation to area size and proximity to source areas, and the role of speciation is also recognized as an important factor. However, one as yet unexplored possibility is that species diversity itself might help to promote speciation, and indeed the central tenets of island biogeographic theory support such a prediction. Here we use data for plants and arthropods of the volcanic archipelagos of the Canary and Hawaiian Islands to address whether there is a positive relationship between species diversity and rate of diversification. Our index of diversification for each island is the proportion of species that are endemic, and we test our prediction that this increases with increasing species number. We show that even after controlling for several important physical features of islands, diversification is strongly related to species number.

Animals↗
PubMed1998

Selenium speciation in human body fluids.

Selenium consumed by humans in foods and in supplements exists in a number of different organic and inorganic forms including selenomethionine, selenocysteine, selenate and selenite. Animal and human studies have established that the bioavailability of the selenium depends upon the chemical form, which also influences the distribution of selenium in the body. These studies have included urinary excretion of selenium following ingestion of different forms of selenium and the response of tissue selenium concentrations and activities of functional selenoproteins to these selenium compounds. Selenomethionine is retained in tissue proteins to a greater extent than selenocysteine and the inorganic forms, but the selenium is not necessarily immediately available for functional selenoproteins. A number of other factors besides chemical form may also influence the bioavailability and distribution of selenium, including other dietary components, selenium status, physiological status and species. Knowledge of these factors and of speciation of selenium in foods, tissues and functional selenoproteins is important for the accurate assessment of selenium status. Speciation of selenium also has implications with respect to the determination of selenium requirements and to the investigation of relationships between selenium status and health and disease.

Biological Availability↗
PubMed1998

Speciation as an analytical aid in trace element research in infant nutrition.

The aim of this work was twofold: to study the binding pattern of trace elements in formulas as compared with breast milk and the relationship between trace elements in breast milk and in maternal dietary intake. To investigate the binding form of trace elements in these nutritive fluids, methods for protein separation were combined with methods for trace element determination in the eluted fractions. HPLC and ICP-AES or ICP-MS were coupled on-line for the simultaneous speciation of elements of nutritional interest, viz., Ca, K, Mg, P, S, Co, Cu, Fe, I, Mn, Mo, Se and Zn, and also the heavy metals Cd and Pb in both human mild whey and formulas. In order to minimize interactions between the labile metal protein complexes and the column material, size-exclusion chromatography was used for protein separation. The binding pattern of trace elements in formulas is significantly different from that in breast milk and depends on its main component (cow's milk or soy), its processing (hydrolysis) and the chemical form (inorganic) of the added compounds. For example, compared with breast-fed infants the iron supply of formula-fed infants is much higher (up to 20-fold); in addition, the binding forms of Fe are very different in the two fluids. This has to be evaluated with respect to interactions with other elements during intestinal uptake. The investigation of breast milk samples from different regions of the world showed comparable shapes for teh elution profiles and for Mo and Se a dependence on the regional maternal dietary intake. Speciation studies carried out on breast milk samples as a function of the selenium content showed significant changes in the zinc-binding pattern. In particular, citrate (as a zinc-binding component) was found to decrease with increasing dietary selenium intake of the mother.

Chromatography, High Pressure Liquid↗
PubMed2000

Speciation of protein-binding zinc and copper in human blood serum by chelating resin pre-treatment and inductively coupled plasma mass spectrometry.

A method for the speciation of zinc and copper binding with proteins in human serum was explored by chelating resin (Chelex-100) pre-treatment and inductively coupled plasma mass spectrometry (ICP-MS). It was shown by a SEC (size-exclusion chromatography)-ICP-MS system that albumin-zinc and albumin-copper (loosely-bound species) could be selectively removed from serum by adsorption on the Chelex-100 resin after the chelating resin pre-treatment, while alpha 2-macroglobulin-zinc and ceruloplasmin-copper (firmly-bound species) remained in the serum. The zinc and copper bound with alpha 2-macroglobulin and ceruloplasmin, respectively, were then determined by ICP-MS after batch treatment of the serum samples with the Chelex-100 resin. In addition, the total concentrations of zinc and copper were also determined by ICP-MS after a 20-fold dilution with 0.1 M HNO3. The albumin-zinc and -copper were estimated as the differences between the concentrations of total and firmly-bound species. The present batch pre-treatment method was applied to the speciation analysis of zinc and copper binding with proteins in sera donated from 25 healthy volunteers as well as from a pregnant woman and a myelodysplastic syndrome patient. The observed concentrations of alpha 2-macroglobulin-zinc and ceruloplasmin-copper were in the ranges 109-202 ng ml-1 (12.4-31.3% of total zinc) and 513-880 ng ml-1 (90.6-99.7% of total copper), respectively. The present method is simple (only addition of the chelating resin and centrifugation is required) and reproducible (average RSD = 2% for alpha 2-macroglobulin-zinc and 1% for ceruloplasmin-copper in intra-assay measurements, and 5% for alpha 2-macroglobulin-zinc and 4% for ceruloplasmin-copper in inter-assay measurements), and there is less risk of contamination during separation.

Blood Proteins↗