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Intersubject variability of risk from perchlorate in community water supplies.

This article is a brief review and summary of the estimated incremental risks (increases in hazard quotient or decreases in thyroid uptake of iodine) to pregnant women (and hence their fetuses) associated with perchlorate exposure in community water supplies (CWSs) . The analysis draws on the recent health effects review published in 2005 by the National Research Council (NRC) . We focus on the potential level of risk borne by the NRC-identified most sensitive subpopulation (pregnant women and hence their fetuses) . Other members of the population should be at a level of risk below that calculated here, and so protection of the sensitive subpopulation would protect the general public health. The analysis examines the intersubject distribution of risks to this sensitive subpopulation at various potential drinking water concentrations of perchlorate and also draws on estimates of the national occurrence of perchlorate in U.S. CWSs to estimate the variability of risks under defined regulatory scenarios. Results suggest that maximum contaminant levels (MCLs) of up to 24.5 microg/L should pose little or no incremental risk to the large majority of individuals in the most sensitive subpopulations exposed in the United States at current levels of perchlorate in water. The protectiveness of an MCL of 24.5 microg/L depends, however, on whether the study subjects in the health effects data used here may be assumed to have been exposed to background (non-drinking water) contributions of perchlorate.

Environmental Exposure↗

Defects in intrathyroid binding of iodine and the perchlorate discharge test.

The kinetics of [123I]iodide uptake were studied when organification of iodine by the thyroid gland was normal and when this binding function was diminished by drugs or disease. Each study was terminated by a sodium perchlorate discharge test (300--600 mg iv) at 60 min or, in some cases, 10--30 min. The results confirmed that binding takes place rapidly in the uninhibited gland with the binding rate constant being at least 0.150 min-1. Discharge from the uninhibited gland is less than 3.5% of the gland uptake when perchlorate is given 60 min after the radioiodide. Subjects with an intrinsic binding defect manifested discharges of 11% of greater of the 60 min uptake and the estimated binding rate constants ranged from 0.003--0.057 min-1. Thyrotoxic subjects receiving 5 mg carbimazole twice daily manifested discharges ranging from 5.4--64.2%, and in those receiving 20 mg twice daily the observed discharges were 67.6--94.6% of the 60 min uptake. The study shows that a correctly performed perchlorate discharge test will detect minimal inhibition of iodine binding. An important factor is the duration of the follow-up period after perchlorate is given. In some of the cases studied discharge was not complete until 60 min after the perchlorate.

Binding Sites↗

Development of freshwater water-quality criteria for perchlorate.

The anion perchlorate (ClO4-) is an oxidizing component commonly used in solid propellants for rockets and missiles; in explosives, flares, fireworks, chemical processes, and automobile air-bag inflators; and for other assorted uses. With recent advances in analytical detection capability, perchlorate has been found in a variety of ground and surface waters throughout the United States. Because perchlorate has been associated with thyroid problems in humans and may have similar effects on wildlife, it is desirable to develop a water-quality criterion to assist in identifying concentrations of perchlorate in water likely to pose an ecological health risk. In the present study, we compiled all available data regarding the effects of perchlorate to aquatic organisms, and we performed additional toxicity and bioconcentration tests as required by the U.S. Environmental Protection Agency (U.S. EPA) for the development of water-quality criteria for aquatic life. A criterion maximum concentration of 20 mg/L and a criterion continuous concentration of 9.3 mg/L were calculated based on the test results. Although these are not formal Clean Water Act Section 304(a) criteria, which must be published by the U.S. EPA, these criteria may be useful in the determination of remedial action levels for contaminated sites, for National Pollutant Discharge Elimination System permit limits, and other water-quality management practices.

Animals↗

Accumulation of perchlorate in aquatic and terrestrial plants at a field scale.

Previous laboratory-scale studies have documented perchlorate ClO(-)(4) uptake by different plant species, but less information is available at field scale, where ClO(-)(4) uptake may be affected by environmental conditions, such as distance to streams or shallow water tables, exposure duration, and species. This study examined uptake of ClO(-)(4) in smartweed (Polygonum spp.) and watercress (Nasturtium spp.) as well as more than forty trees, including ash (Fraxinus greggii A. Gray), chinaberry (Melia azedarach L.), elm (Ulmus parvifolia Jacq.), willow (Salix nigra Marshall), mulberry [Broussonetia papyrifera (L.) Vent.], and hackberry (Celtis laevigata Willd.) from multiple streams surrounding a perchlorate-contaminated site. Results indicate a large potential for ClO(-)(4) accumulation in aquatic and terrestrial plants, with ClO(-)(4) concentration in plant tissues approximately 100 times higher than that in bulk water. Perchlorate accumulation in leaves of terrestrial plants was also dependent on species, with hackberry, willow, and elm having a strong potential to accumulate ClO(-)(4). Generally, trees located closer to the stream had a higher ClO(-)(4) accumulation than trees located farther away from the stream. Seasonal leaf sampling of terrestrial plants indicated that ClO(-)(4) accumulation also was affected by exposure duration, with highest accumulation observed in the late growing cycle, although leaf concentrations for a given tree were highly variable. Perchlorate may be re-released into the environment via leaching and rainfall as indicated by lower perchlorate concentrations in collected leaf litter. Information obtained from this study will be helpful to understand the fate of ClO(-)(4) in macrophytes and natural systems.

Nasturtium↗

Analysis of perchlorate in water and soil by electrospray LC/MS/MS.

A method has been developed for the determination of perchlorate in water and soil matrixes using electrospray liquid chromatography/mass spectrometry/mass spectrometry. Perchlorate is quantitated by monitoring the ion signal from mass 83, which is formed by a loss of an oxygen atom from the perchlorate molecular ion. The method was developed to be effective and economical in production laboratory analysis of perchlorate in environmental water and soil samples. Data were gathered to define method sensitivity, performance, selectivity, and robustness. Analyte stability, method susceptibility to interferences, and the reliability of the chlorine isotope ratio as an identification tool were examined. The aqueous method detection limit (MDL) is 0.05 microg/L and was determined using an actual groundwater matrix. The soil MDL is 0.5 microg/kg and was determined using Ottawa sand. The stability study was performed by spiking water samples at 0.25, 10, and 20 microg/L and analyzing them 50 days later. Acceptable recoveries were obtained for all samples. The relative standard deviation (RSD) for the replicate analyses in the stability study indicates that the method is capable of RSD values less than 5% in a relatively clean groundwater matrix. The ionization suppression study was performed by spiking water samples containing 1000 mg/L carbonate, chloride, and sulfate with 0.05 and 0.5 microg/L perchlorate and then measuring the recovery of the spike. The results indicate that the procedure does not have significant suppression effects at the high salt levels tested. Calibration, quality control sample, field sample, and suppression study data were combined to examine isotope ratio reliability. The results of that work show that chlorine isotope ratios can be used to define statistical process control limits for use as an additional analyte identification tool.

Journal Article↗

Dracorhodin perchlorate induces apoptosis via activation of caspases and generation of reactive oxygen species.

Dracorhodin perchlorate inhibited proliferation of several tumor cell lines. The drug induced oligonucleosomal fragmentation of DNA in HeLa cells and increased caspase-3, -8, -9 activities followed by the degradation of caspase-3 substrates, inhibitor of caspase-dependent DNase, and poly-(ADP-ribose) polymerase. It also increased caspase-1 activity and a caspase-1 inhibitor, Ac-YVAD-cmk, and a caspase-10 inhibitor z-AEVD-fmk, also reduced dracorhodin-perchlorate-induced HeLa cell death. Dracorhodin perchlorate decreased the expression of anti-apoptotic mitochondrial protein, Bcl-X(L), but not Bcl-2; and it increased the expression of pro-apoptotic protein, Bax. Dracorhodin perchlorate induced a sustained generation of reactive oxygen species (ROS) in HeLa cells; caspase-1 inhibitor, Ac-YVAD-cmk, and caspase-3 inhibitor, z-DEVD-fmk, attenuated the generation of ROS. Taken together, our results indicate that dracorhodin perchlorate alters the intracellular redox status, changed the balance of Bcl-X(L) and Bax protein expression, and induces apoptosis through caspase pathways in HeLa cells.

Antimetabolites, Antineoplastic↗

Accelerated fall in serum bromide level after administration of perchlorate to man.

Five patients intoxicated with bromoureide showed accelerated elimination of bromide from serum after a single dose of perchlorate. It was probably due to increased renal excretion of bromide. Perchlorate has been reported to increase the renal clearance of other monovalent anions by inhibiting their tubular reabsorption and a similar interaction between perchlorate and bromide is suggested. Although perchlorate effectively eliminates bromide it should not replace chloride in the treatment of bromide intoxication, because of the side effects of its repeated administration.

Adult↗

Interaction between perchlorate and nifedipine on insulin secretion from mouse pancreatic islets.

In order to elucidate the mechanisms responsible for the stimulatory effect of perchlorate (ClO4-) on insulin secretion, we have investigated the interaction between this chaotropic anion and the organic calcium antagonist nifedipine. This drug, known as a blocker of L-type calcium channels, was chosen as a tool to test the idea that ClO4- acts on insulin secretion by stimulating the gating of voltage-controlled Ca2+ channels. ClO4- amplified the stimulatory effect of D-glucose on insulin release from perfused pancreas (first and second phases) as well as from isolated islets incubated in static incubations for 60 min. This indicates that ClO4- amplifies physiologically regulated insulin secretion. Nifedipine reduced D-glucose-induced (20 mM) insulin release in a dose-dependent manner with half-maximum effect at about 0.8 microM and apparent maximum effect at 5 microM nifedipine. In the presence of 20 mM D-glucose, the inhibitory effects of 0.5, 1 or 5 microM nifedipine were only slightly, if at all, counteracted by perchlorate. When 12 mM ClO4- and 20 mM D-glucose were combined, calculation of the specific effect of ClO4- revealed that nifedipine produced almost maximum inhibition already at 0.05 microM. Thus, the perchlorate-induced amplification of D-glucose-stimulated insulin release shows higher sensitivity to nifedipine than the D-glucose-effect as such. This supports the hypothesis that perchlorate primarily affects the voltage-sensitive L-type calcium channel in the beta-cell.

Animals↗

Uptake, accumulation and depuration of sodium perchlorate and sodium arsenate in zebrafish (Danio rerio).

In toxicokinetics studies, interactions between chemicals in mixtures has been largely neglected. This study examines a mixture of perchlorate and arsenate because (1) they have the potential to co-occur in contaminated aquatic habitats, and (2) a previous study by the authors found possible toxicological interactive effects. In the present study, zebrafish (Danio rerio) were exposed to two concentrations of sodium perchlorate (10 and 100 mg l(-1)), sodium arsenate (1 and 10 mg l(-1)), and the mixture-sodium perchlorate+sodium arsenate (10+1 mg l(-1) and 100+10 mg l(-1) Na(2)HAsO(4)-high mixture) for 90 d. Their uptake and accumulation by zebrafish was evaluated at 10, 30, 60, and 90 d. In addition, depuration was examined at 1, 3, and 5d after cessation of the exposure. The uptake of either chemical was concentration-dependent, with significantly higher uptake at high concentrations at either exposure interval. In contrast, there was no significant difference in whole body residue between single chemicals and the corresponding mixture except for 100 mg l(-1) sodium arsenate at 90 d. However, there was increasing accumulation over time at the high concentration of either chemical alone and their mixture, and this increasing trend was more pronounced in the single chemical exposures than in the mixture. At the concentrations tested in the current study, both chemicals reduced the uptake but enhanced the depuration of the other chemical from the zebrafish. This study represents the first examination of the interaction of two anions-perchlorate and arsenate with respect to toxicokinetics.

Animals↗

Optimization of operating conditions for the determination of perchlorate in biological samples using preconcentration/preelution ion chromatography.

Perchlorate originates as a contaminant in the environment from the use of salts in the manufacture of solid rocket fuels and munitions. Monitoring potential perchlorate contamination in the environment is of interest, however, very few analytical methods have been developed for perchlorate determination in biological samples. Analysis of complex samples by ion chromatography is complicated by matrix components that can interfere with perchlorate determination. However, a recently developed preconcentration/preelution (PC/PE) ion chromatography method has demonstrated the capability to analyze certain complex samples such as high salinity water, milk, and hydroponic fertilizers. The ability of this method to reduce sample background and lower detection limits in ion chromatography for various biological samples was evaluated in this study. The PC/PE method was applicable to the analysis of kidneys, livers, zebrafish, quail eggs, lettuce, and urine. Optimal operating conditions were determined for each matrix. Ranges of optimal wash volumes were shorter when 15 mM NaOH prewash solutions were used compared with 10mM and good recovery was achieved for most matrices with an injection period > or =60s. Prewash solution concentration did not appear to significantly affect matrix background. The PC/PE method was capable of reducing sample background when compared to EPA Method 314.0, which resulted in detection limits, with the exception of zebrafish and urine, that were two-fold lower than those achieved with EPA Method 314.0.

Animals↗

Effect of perchlorate on the fluorescence of protonated 2,2'-bipyridine.

Interaction between the monocation of an aromatic amine, 2,2'-bipyridine and perchlorate anion was studied. The protonation process by perchloric acid has been followed by the changes in the UV absorption spectra and in the fluorescence emission. Addition of lithium perchlorate with increasing amount results in a significant increase in the fluorescence of 2,2'-bipyridine (the bipyridyl cation). This increase in fluorescence indicates that the rotation in the protonated monocation is hindered in the presence of perchlorate. We assume that this hindrance is due to the formation of the BpH+.ClO4- ion-pair.

2,2'-Dipyridyl↗

Interspecies differences in susceptibility to perturbation of thyroid homeostasis: a case study with perchlorate.

Despite many physiological similarities, humans and rats exhibit notably different susceptibilities to thyroid perturbation. Considerable research has recently been conducted on the thyroid-active chemical perchlorate, a chemical of emerging environmental and regulatory interest. While the data indicate humans and rats exhibit similar dose-response relationships in terms of acute inhibition of thyroidal iodide uptake, the two species appear to exhibit notable differences in terms of thyroid hormone response, the toxicologically significant consequence of iodide uptake inhibition. We analyzed dose-response data for changes in serum T(3), T(4), and TSH levels from studies in humans, rats, mice, and rabbits. We found that thyroid homeostasis in the rat appears to be strikingly more sensitive to perchlorate than any of the other species. Rats exhibited an increase in serum TSH at 0.1mg/kg-day whereas other species remained unresponsive even at doses of 10mg/kg-day. Less pronounced but consistent effects were seen with serum T(3) and T(4). These cross-species comparisons provide strong evidence that data obtained from rat studies should be critically evaluated for their relevance to humans. If rat data are used to develop toxicity criteria for perchlorate, we propose that this is an instance where an inter-species uncertainty factor less than one is supportable. DISCLOSURE STATEMENT: One of the authors (BDB) has been hired by Lockheed Martin Corporation as an expert in litigation involving perchlorate. A portion of the initial research presented in this paper was conducted in conjunction with her role in that matter.

Animals↗

Conformational transitions of alternating purine-pyrimidine DNAs in perchlorate ethanol solutions.

Conformational transitions of poly(dA-dC).poly(dG-dT), poly(dA-dT).poly(dA-dT), and other alternating purine-pyrimidine DNAs were studied in aqueous ethanol solutions containing molar concentrations of sodium perchlorate, which is a novel solvent stabilizing non-B duplexes of DNA. Using CD and UV absorption spectroscopies, we show that this solvent unstacks bases and unwinds the B-forms of the DNAs to transform them into the A-form or Z-form. In the absence of divalent cations poly(dA-dC).poly(dG-dT) can adopt both of these conformations. Its transition into the Z-form is induced at higher salt and lower ethanol concentrations, and at higher temperatures than the transition into the A-form. Submillimolar concentrations of NiCl2 induce a highly cooperative and slow A-Z transition or Z-Z' transition, which is fast and displays low cooperativity. Poly(dA-dT).poly(dA-dT) easily isomerizes into the A-form in perchlorate-ethanol solutions, whereas high perchlorate concentrations denature the polynucleotide, which then cannot adopt the Z-form. At low temperatures, however, NiCl2 also cooperatively induces the Z'-form in poly(dA-dT).poly(dA-dT). Poly(dI-dC).poly(dI-dC) is known to adopt an unusual B-form in low-salt aqueous solution, which is transformed into a standard B-form by the combination of perchlorate and ethanol. NiCl2 then transforms poly(dI-dC).poly(dI-dC) into the Z'-form, which is also adopted by poly(dI-br5dC).poly(dI-br5dC).

Base Sequence↗

Determination of perchlorate at parts-per-billion levels in plants by ion chromatography.

A method for the analysis of perchlorate in plants was developed, based on dry weight, and applied to the analysis of plant organs, foodstuffs, and plant products. The method reduced greatly the ionic interferences in water extracts of plant materials. The high background conductivity, due to the plant matrix, was reduced sufficiently to allow quantitation of perchlorate with little or no matrix interference. Ion chromatography (IC) on a microbore AS16 anion-exchange column and a conductivity detector was used for separation and detection of perchlorate from the ionic plant extract. The extract was heated to precipitate proteins, centrifuged, exposed to alumina, and filtered through a cartridge filled with divinylbenzene to yield a water clear extract for IC analysis, even from highly colored solutions. Heating the extract and treatment with alumina reduced substantially the ionic content of the extracts without loss of perchlorate.

Chromatography, Liquid↗

Determination of trace level bromate and perchlorate in drinking water by ion chromatography with an evaporative preconcentration technique.

A simple sample preconcentration technique employing microwave-based evaporation for the determination of trace level bromate and perchlorate in drinking water with ion chromatography is presented. With a hydrophilic anion-exchange column and a sodium hydroxide eluent in linear gradient, bromate and perchlorate can be determined in one injection within 35 min. Prior to ion chromatographic analysis, the drinking water sample was treated with an OnGuard-Ag cartridge to remove the superfluous chloride and concentrated 20-fold using a PTFE beaker in a domestic microwave oven for 15 min. The recoveries of the anions ranged from 94.6% for NO2- to 105.2% for F-. The detection limits for bromate, perchlorate, iodate and chlorate were 0.1, 0.2, 0.1 and 0.2 microg/l, respectively. The developed method is applicable for the quantitation of bromate and perchlorate in drinking water samples.

Bromates↗

Study of tryptophan enantiomer binding to a teicoplanin-based stationary phase using the perturbation technique. Investigation of the role of sodium perchlorate in solute retention and enantioselectivity.

The retention of D,L-tryptophan enantiomers on an immobilized teicoplanin column was investigated in relation to the mobile phase sodium perchlorate concentration using the perturbation method to determine the solute distribution isotherms. From the experimental data, it appeared that the bi-Langmuir model was able to describe D- and L-enantiomer retention on the immobilized selector over the salt concentration range. An increase in the apparent enantioselectivity with an increase in sodium perchlorate concentration was observed. The chiral recognition enhancement was governed by (i) an increase in the difference of the adsorption constants for binding to the high-affinity site (aglycone pocket) between the two enantiomers and (ii) enhancement of the number of aglycone chiral regions interacting with D-tryptophan. It is suggested that an ion-pair formation mechanism between perchlorate and solute and/or selector is responsible for this behavior. In addition, this work shows that additional secondary sites on the teicoplanin surface are involved in the apparent enantioselectivity at low sodium perchlorate concentrations.

Perchlorates↗

Determination of trace levels of haloacetic acids and perchlorate in drinking water by ion chromatography with direct injection.

Disinfection by products of haloacetic acids and perchlorate pose significant health risks, even at low microg/l levels in drinking water. A new method for the simultaneous determination of nine haloacetic acids (HAAs) and perchlorate as well as some common anions in one run with ion chromatography was developed. The HAAs tested included mono-, di-, trichloroacetic acids, mono, di-, tribromoacetic acids, bromochloroacetic acid, dibromochloroacetic acid, and bromodichloroacetic acid. Two high-capacity anion-exchange columns, a carbonate-selective column and a hydroxide-selective hydrophilic one, were used for the investigation. With the carbonate-selective column, the nine HAAs as well as fluoride, chloride, nitrite, nitrate, phosphate and sulfate could be well separated and determined in one run. With the very hydrophilic column and a gradient elution of sodium hydroxide, methanol and deionized water, the nine HAAs, fluoride, chloride, nitrite, nitrate as well as perchlorate could be simultaneously determined in one run within 34 min. The detection limits for HAAs were between 1.11 and 9.32 microg/l. For perchlorate, it was 0.60 microg/l.

Acetates↗

Perchlorate levels in samples of sodium nitrate fertilizer derived from Chilean caliche.

Paleogeochemical deposits in northern Chile are a rich source of naturally occurring sodium nitrate (Chile saltpeter). These ores are mined to isolate NaNO3 (16-0-0) for use as fertilizer. Coincidentally, these very same deposits are a natural source of perchlorate anion (ClO4-). At sufficiently high concentrations, perchlorate interferes with iodide uptake in the thyroid gland and has been used medicinally for this purpose. In 1997, perchlorate contamination was discovered in a number of US water supplies, including Lake Mead and the Colorado River. Subsequently, the Environmental Protection Agency added this species to the Contaminant Candidate List for drinking water and will begin assessing occurrence via the Unregulated Contaminants Monitoring Rule in 2001. Effective risk assessment requires characterizing possible sources, including fertilizer. Samples were analyzed by ion chromatography and confirmed by complexation electrospray ionization mass spectrometry. Within a lot, distribution of perchlorate is nearly homogeneous, presumably due to the manufacturing process. Two different lots we analyzed differed by 15%, containing an average of either 1.5 or 1.8 mg g-1. Inadequate sample size can lead to incorrect estimations; 100-g samples gave sufficiently consistent and reproducible results. At present, information on natural attenuation, plant uptake, use/application, and dilution is too limited to evaluate the significance of these findings, and further research is needed in these areas.

Chile↗