Chromium speciation and biochemical changes vary in relation to plant ploidy.

Uptake of trivalent chromium (Cr(III)-chloride), Cr speciation and consequences for the metabolism in chamomile plants with two ploidy levels have been studied. Depletion of fresh biomass, tissue water content and soluble proteins in response to high (120 µM) Cr(III) was ploidy-independent. Cr mainly accumulated in the roots (only negligibly in the shoots) and total root Cr amount was higher in tetraploid ones including the proof with specific fluorescent indicator (naphthalimide-rhodamine) of Cr(III). Quantification of Cr(VI) detected its higher content in tetraploid roots (up to 4.2% from total Cr), indicating partial oxidation of applied Cr(III). Higher H2O2 presence but lower activities of peroxidases were observed in tetraploid roots while nitric oxide, superoxide dismutase and glutathione reductase activities did not differ extensively. Soluble phenols, lignin, non-protein thiols, individual thiols (glutathione and phytochelatin 2) and ascorbic acid responded to high Cr(III) similarly in both cultivars while decrease of minerals was more pronounced in tetraploid ones. It seems that Cr(III)-induced oxidative stress arises from high root Cr uptake and Cr(VI) presence and is related to depletion of thiols. Assay of Krebs cycle acids confirmed rather depletion under 120 µM Cr(III) in both cultivars but increase in citric acid may indicate its involvement in root Cr chelation. Subsequent comparison of Cr(III)-chloride and Cr(III)-nitrate showed similar influence on Cr accumulation and majority of biochemical responses while different impact on phytochelatin 2 amount was the most distinct feature.

Chromium and its speciation in water samples by HPLC/ICP-MS--technique establishing metrological traceability: a review since 2000.

Chromium holds a special position among living organisms because depending on its species it can be either essential or toxic. Cr(VI) even at very low concentrations is harmful and carcinogenic, while Cr(III) is a necessary microelement for cellular metabolism. Therefore, a simple analysis of Cr concentration in collected samples will not be able to distinguish these differences effectively: for a proper chemical analysis we need to perform a reliable detection and quantification of Cr species. Separation and detection of chromium can be accomplished with high performance liquid chromatography hyphenated to inductively coupled plasma mass spectrometry (HPLC/ICP-MS) in a one-step. Our review assembles articles published since 2000 regarding chromium speciation in water samples with the use of HPLC/ICP-MS. It addresses the following issues: chromium chemistry, the possibilities of dealing with interferences, metrological aspects, analytical performance and speciated isotope dilution mass spectrometry (SIDMS) which is a definitive measurement method. The authors would like to advocate this hyphenated advanced technique as well as the metrological approach in speciation analysis of chromium.

Speciation of chromium using chronoamperometric biosensors based on screen-printed electrodes.

Chronoamperometric assays based on tyrosinase and glucose oxidase (GOx) inactivation have been developed for the monitoring of Cr(III) and Cr(VI). Tyrosinase was immobilized by crosslinking on screen-printed carbon electrodes (SPCEs) containing tetrathiafulvalene (TTF) as electron transfer mediator. The tyrosinase/SPC(TTF)E response to pyrocatechol is inhibited by Cr(III). This process, that is not affected by Cr(VI), allows the determination of Cr(III) with a capability of detection of 2.0±0.2 µM and a reproducibility of 5.5%. GOx modified screen-printed carbon platinised electrodes (SPCPtEs) were developed for the selective determination of Cr(VI) using ferricyanide as redox mediator. The biosensor was able to discriminate two different oxidation states of chromium being able to reject Cr(III) and to detect the toxic species Cr(VI). Chronoamperometric response of the biosensor towards glucose decreases with the presence of Cr(VI), with a capability of detection of 90.5±7.6 nM and a reproducibility of 6.2%. A bipotentiostatic chronoamperometric biosensor was finally developed using a tyrosinase/SPC(TTF)E and a GOx/SPC(Pt)E connected in array mode for the simultaneous determination of Cr(III) and Cr(VI) in spiked tap water and in waste water from a tannery factory samples.

Assessment of genotoxic potential of Cr(VI) in the mouse duodenum: an in silico comparison with mutagenic and nonmutagenic carcinogens across tissues.

In vitro studies on hexavalent chromium [Cr(VI)] indicate that reduced forms of this metal can interact with DNA and cause mutations. Recently, Cr(VI) was shown to induce intestinal tumors in mice; however, Cr(VI) elicited redox changes, cytotoxicity and hyperplasia - suggesting involvement of tissue injury rather than direct mutagenesis. Moreover, toxicogenomic analyses indicated limited evidence for DNA damage responses. Herein, we extend these toxicogenomic analyses by comparing the gene expression patterns elicited by Cr(VI) with those of four mutagenic and four nonmutagenic carcinogens. To date, toxicogenomic profiles for mutagenic and nonmutagenic duodenal carcinogens do not exist, thus duodenal gene changes in mice were compared to those elicited by hepatocarcinogens. Specifically, duodenal gene changes in mice following exposure to Cr(VI) in drinking water were compared to hepatic gene changes previously identified as potentially discriminating mutagenic and nonmutagenic hepatocarcinogens. Using multivariate statistical analyses (including logistic regression classification), the Cr(VI) gene responses clustered apart from mutagenic carcinogens and closely with nonmutagenic carcinogens. These findings are consistent with other intestinal data supporting a nonmutagenic mode of action (MOA). These findings may be useful as part of a full weight of evidence MOA evaluation for Cr(VI)-induced intestinal carcinogenesis. Limitations to this analysis will also be discussed.

Sequential eluent injection technique as a new approach for the on-line enrichment and speciation of Cr(III) and Cr(VI) species on a single column with FAAS detection.

This paper introduces a sequential eluent injection (SEI) technique combined with an on-line preconcentration/separation system for a fast and sensitive FAAS determination of trace amounts of Cr(III) and Cr(VI) species. The method is based on the simultaneous retention of Cr(III) and Cr(VI) on a single mini-column packed with a chloromethylated polystyrene functionalized with N,N-bis(naphthylideneimino)diethylenetriamine (PS-NAPdien) at pH 6.7. The retained chromium species was eluted by sequential injection of HCl for desorption of Cr(III), and NH(3) and NH(4)NO(3) buffer solutions for desorption of Cr(VI). All the chemical and flow injection variables were optimized for the quantitative preconcentration and speciation of Cr(III) and Cr(VI). Under the optimum conditions, the calibration graph obtained is linear over the concentration range of 2.0-60.0µg L(-1) for Cr(III), and 8.0-180.0µg L(-1) for Cr(VI). The preconcentration factors for Cr(III) and Cr(VI) were 70 and 30, respectively. The 3σ detection limits were 0.6µg L(-1) and 2.5µg L(-1) for Cr(III) and Cr(VI), respectively. The relative standard deviations were 2.55% and 0.8%, respectively, for 6 replicate determinations of Cr(III) and Cr(VI) at the 40.0µg L(-1) level. The proposed method was applied for determination of Cr(III) and Cr(VI) in different water samples with satisfactory results.

Chromium speciation in a contaminated groundwater: redox processes and temporal variability.

Chromium species (Cr(III), Cr(VI), and Cr(III)-organic) in groundwater of a tannery contaminated area were monitored during pre- and post-monsoon seasons for a period of 3 years (May 2004 to January 2007). The objectives of the study were (1) to investigate the temporal variation of chromium species and other matrix constituents and (2) to study the redox processes associated with the temporal variation of chromium species. Samples were collected from 15 dug wells and analyzed for chromium species and other constituents. The results showed that the groundwater was relatively more oxidizing during post-monsoon periods than the pre-monsoon periods. Except one sample, the concentration of chromium species were found in the order of Cr(VI)>Cr(III)>Cr(III)-organic complexes during all the pre- and post-monsoon periods. In most of the wells, the concentrations of Cr(III), Cr(VI), and Cr(III)-organic decreased during post-monsoon periods compared to their pre-monsoon concentrations. However, the Cr(VI)/Cr(Total) ratio still increased and the Cr(III)/Cr(Total) ratio decreased during post-monsoon periods in most of the samples. The possible mechanisms for the temporal variation of chromium species were (1) Fe(II) reduction of Cr(VI) vs oxidation of Fe(II) by dissolved oxygen and (2) oxidation of Cr(III) by Mn(IV).

Pyridine appended L-methionine: a novel chelating resin for pH dependent Cr speciation with scanning electron microscopic evidence and monitoring of yeast mediated green bio-reduction of Cr(VI) to Cr(III) in environmental samples.

Chemical speciation and pH dependent separation of Cr(III) and Cr(VI) species in environmental samples have been achieved by solid phase extraction using a new chelating resin containing pyridine appended L-methionine. Cr(III) is completely sorbed on the resin at pH 8.0 and Cr(VI) at pH 2.0. Hence a pH dependent separation of Cr(III) and Cr(VI) is possible with a limit of detection of 1.6 µg mL(-1) and 0.6 µg mL(-1) respectively. The sorption capacity of the resin for Cr(III) and Cr(VI) is 2.8 mmol g(-1) and 1.3 mmol g(-1) respectively. The sorption of chromium on the resin is supported by scanning electron microscopy (SEM). Complete desorption of Cr(III) and Cr(VI) from 1g of Cr loaded resin was achieved using 10 mL of 2 mol L(-1) HNO(3) and 6 mL of 3 mol L(-1) HNO(3) respectively. Quantitative recoveries of Cr(III) (pH 8.0) and Cr(VI) (pH 2.0) were found to be 96.0% and 98.0% respectively. Reduction efficiency of Rhodotornula mucilaginosa yeast from Cr(VI) to Cr(III) was monitored with this new resin. Concentrations of metal ions were measured by flame atomic absorption spectroscopy (FAAS).

Speciation of chromium metal ions by RP-HPLC.

A high-performance liquid chromatographic method with UV detection, based on chelation with morpholine-4-carbodithioate (MDTC), has been developed for the speciation of chromium. Simultaneous preconcentration of Cr(III) and Cr(VI) in aqueous solutions was achieved by extraction of metal species in chloroform as MDTC complexes. The separation of MDTC chelates of Cr(III) and Cr(VI) was realized on a C18 column with the use of acetonitrile-water (70:30) at a flow rate of 1.0 mL/min. The Cr(III) showed a single peak in chromatogram due to Cr(MDTC)(3), and Cr(VI) showed two peaks due to the formation of Cr(MDTC)(3) and Cr(MDTC)(2)(OMDTC). These two complexes were separated with different retention times at 5.4 and 4.3 min, respectively. The developed method can be used for rapid routine determination of chromium species with high precision and reliability. The method was validated by applying to various synthetic binary mixtures prepared by spiking Cr(III) and Cr(VI) in tap water.

Immobilized chitosan as biosorbent for the removal of Cd(II), Cr(III) and Cr(VI) from aqueous solutions.

The generation of layer-by-layer silicate-chitosan composite biosorbent was studied. The films were evaluated on its stability regarding the polymer leakage and its capability in the removal of Cd(II), Cr(III) and Cr(VI) from an aqueous solution. SEM, EDAX and ATR-IR techniques were applied for material characterization. Silicate-chitosan films with a final layer of silicate demonstrated chitosan retention and had better sorption capacities than those without it. For metal species, such as Cd(II) and Cr(III), the greatest adsorption was obtained when the pH of the solution was 7. When Cr(VI) was evaluated, pH 4 was the optimal for its adsorption. Langmuir and Freundlich isotherms were modeled for the equilibrium data. An 80% of the adsorbed metal was recovered by HNO(3) incubation. This non-covalent immobilization method allowed chitosan surface retention and did not affect its adsorption properties. The use of a coated surface would facilitate sorbent removal from medium after adsorption.

Use of micro-XANES to speciate chromium in airborne fine particles in the Sacramento Valley.

While particulate matter (PM) in the atmosphere can lead to a wide array of negative health effects, the cause of toxicity is largely unknown. One aspect of PM that likely affects health is the chemical composition, in particular the transition metals within the particles. Chromium is one transition metal of interest due to its two major oxidation states, with Cr(III) being much less toxic compared to Cr(VI). Using microfocused X-ray absorption near edge structure (micro-XANES), we analyzed the Cr speciation in fine particles (diameters < or = 2.5 microm) collected at three sites in the Sacramento Valley of northern California. The microfocused X-ray beam enables us to look at very small areas on the filter with a resolution of typically 5-7 micrometers. With XANES we are able to not only distinguish between Cr(VI) and Cr(III), but also to identify different types of Cr(III) and more reduced Cr species. At all of our sampling sites the main Cr species were Cr(III), with Cr(OH)3 or a Cr-Fe, chromite-like, phase being the dominant species. Cr(VI)-containing particles were found only in the most urban site. All three sites contained some reduced Cr species, either Cr(0) or Cr3C2, although these were minor components. This work demonstrates that micro-XANES can be used as a minimally invasive analytical tool to investigate the composition of ambient PM.

Speciation of chromium by in-capillary derivatization and electrophoretically mediated microanalysis.

An electrophoretic method for chromium speciation analysis--as Cr(III) and Cr(VI)--based on in-capillary derivatization with 1,5-diphenylcarbazide (DPC) is here proposed. As Cr(III) does not react with DPC, it was oxidized also in-capillary to Cr(VI) by Ce(IV). For this purpose, a capillary electrophoresis (CE) mode called electrophoretically mediated microanalysis (EMMA) based on sequential injection of sample and reagents--namely, DPC, sample and Ce(IV)--was employed. The conditions of both reactions--Cr(III) oxidation and Cr(VI)-DPC derivatization--were optimized in order to quantify separately the Cr(VI)-DPC complex from the original Cr(VI) in the sample and that from oxidation of Cr(III) to Cr(VI). The electrophoretic conditions were independently optimized for variables influencing the resolution and those affecting sensitivity. The method thus developed was applied to the determination of Cr(III) and Cr(VI) in glass material, for which different sample preparation methods--namely, EPA method 3060A, ultrasound-assisted leaching and microwave-assisted digestion--were tested. Microwave-assisted digestion was found to be the best sample preparation alternative in terms of efficiency of the step--99.6 and 98.3% for Cr(VI) and Cr(III), respectively--and procedure time--20 min. The complete method was validated with the certified reference material BAM-S004.

New combination of EXAFS spectroscopy and density fractionation for the speciation of chromium within an andosol.

Studying speciation of heavy metals instead of their total concentration in a complex matrix such as soil is a scientific challenge that requires a combination of different analytical techniques. We compared the speciation of Cr within an andosol (island of Réunion) by using X-ray absorption spectroscopy (XAS) and sequential extraction. Contradictory results were obtained since the fraction of Cr bound to organic matter was detected only with the latter method. As bulk soil is rather complex, its fractionation by a densimetric method decreased its heterogeneity. We found that 60% Cr was within chromite-type primary minerals. Weathering of these phases led to Fe and Cr release, eventually resulting in either coprecipitation as mixed FeCr oxyhydroxide (16% total Cr) or precipitation of a Cr oxyhydroxide (5% total Cr). Our results also revealed that 13% Cr was bound to organic matter. The organic matter was complexed with mineral phases to form organomineral complexes with a density ranging from 1.9 to 2.6. The use of an original density fractionation-based sample preparation allowed identification of the role of organic matter in chromium speciation within an andosol and to overcome the difficulties of EXAFS to detect light elements in the vicinity of heavy elements.

Chromium speciation in environmental samples by solid phase extraction on Chromosorb 108.

In the present work, a solid phase extraction system has been proposed for speciation of Cr(III) and Cr(VI) in the real samples. The procedure based on the adsorption of chromium(III) as dithizonate chelate on the Chromosorb 108 resin. After reduction of Cr(VI) by concentrated H2SO4 and ethanol, the system was applied to the total chromium. Cr(VI) was calculated as the difference between the total Cr content and the Cr(III) content. The influences of the analytical parameters including pH of the aqueous solution, amounts of dithizone, eluent type, sample volume and flow rates of the sample and eluent solution were investigated. No considerable interferences have been observed from other investigated anions and cations on the chromium speciation. The adsorption capacity of sorbent was 4.50 mg/g Cr(III). The detection limit of Cr(III) is 0.75 microg/L. The proposed method was applied to the speciation of chromium in environmental samples including natural waters and total chromium preconcentration in microwave digested Turkish tobacco, coffee and soil samples with satisfactory results. In order to verify the accuracy of the method, two certified reference materials (NIST SRM 1573a Tomato Leaves and RTC-CRM 025-050 Metals on Soil) were analyzed and the results obtained were in good agreement with the certified values. The relative errors and relative standard deviations were below 5% and 9%, respectively.

Dental devices; dental noble metal alloys and dental base metal alloys; designation of special controls. Final rule.

The Food and Drug Administration is amending the identification and classification regulations of gold-based alloys and precious metal alloys for clinical use and base alloys devices in order to designate a special control for these devices. FDA is also exempting these devices from premarket notification requirements. The agency is taking this action on its own initiative. This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Safe Medical Devices Act of 1990 (SMDA), and the Food and Drug Administration Modernization Act of 1997 (FDAMA). Elsewhere in this issue of the Federal Register, FDA is announcing the availability of the draft guidance documents that would serve as special controls for these devices.

Speciation of Cr(III) and Cr(VI) in potable waters by using activated neutral alumina as collector and ET-AAS for determination.

The speciation of Cr(III) and Cr(VI) has been performed by using activated neutral alumina as adsorbent. Both species were quantitatively adsorbed on a small column filled with neutral alumina. The adsorbed Cr(III) was eluted with 4 mol L(-1) HNO(3) and Cr(VI) with 1.0 mol L(-1) ammonia solution. Recoveries of Cr(III) and Cr(VI) were 99% and 100%, respectively. Using ET-AAS for Cr determination the limit of detection in the sample was 0.01 microg L(-1). The combined procedure is fast and sensitive. It can be applied for routine analysis of water samples at sub-microg L(-1) levels with a relative standard deviation (RSD) of 2-10% (three determinations).

Suicide by ingestion of a CCA wood preservative.

Chromated-copper-arsenate (CCA) is a compound used worldwide for wood preservation. Occupational hazards from chronic exposure to CCA are well known, but acute ingestion of CCA wood preservative is very rare. We describe a case of suicide by ingestion of a CCA wood preservative. A 33-year-old man attempted suicide by ingesting an unknown liquid, later identified as a CCA wood preservative, 75 min before his arrival in the emergency department. He was in severe respiratory distress, drooling, tachycardic, and hypotensive. There was an orange color on the palms of both hands. Severe, green colored burns of the buccal mucosa were observed. He was intubated shortly after arrival. The larynx was edematous, but a 7-mm endotracheal tube was successfully introduced. Blood tests revealed partially compensated metabolic acidosis. The patient deteriorated rapidly; the systolic blood pressure dropped to 70 mmHg, and he passed bloody diarrhea. He developed multiple premature atrial contractions and supraventricular tachycardia, and later in the intensive care unit, refractory ventricular tachycardia and ventricular fibrillation. The patient was declared dead 212 h after his arrival. Unfortunately, postmortem blood levels of heavy metals and autopsy were not performed because of refusal by the family for religious reasons.