Erik Huusfeldt Larsen

Distribution of silver in rats following 28 days of repeated oral exposure to silver nanoparticles or silver acetate

BackgroundThe study investigated the distribution of silver after 28 days repeated oral administration of silver nanoparticles (AgNPs) and silver acetate (AgAc) to rats. Oral administration is a relevant route of exposure because of the use of silver nanoparticles in products related to food and food contact materials.ResultsAgNPs were synthesized with a size distribution of 14 ± 4 nm in diameter (90% of the nanoparticle volume) and stabilized in aqueous suspension by the polymer polyvinylpyrrolidone (PVP). The AgNPs remained stable throughout the duration of the 28-day oral toxicity study in rats. The organ distribution pattern of silver following administration of AgNPs and AgAc was similar. However the absolute silver concentrations in tissues were lower following oral exposure to AgNPs. This was in agreement with an indication of a higher fecal excretion following administration of AgNPs. Besides the intestinal system, the largest silver concentrations were detected in the liver and kidneys. Silver was also found in the lungs and brain. Autometallographic (AMG) staining revealed a similar cellular localization of silver in ileum, liver, and kidney tissue in rats exposed to AgNPs or AgAc.Using transmission electron microscopy (TEM), nanosized granules were detected in the ileum of animals exposed to AgNPs or AgAc and were mainly located in the basal lamina of the ileal epithelium and in lysosomes of macrophages within the lamina propria. Using energy dispersive x-ray spectroscopy it was shown that the granules in lysosomes consisted of silver, selenium, and sulfur for both AgNP and AgAc exposed rats. The diameter of the deposited granules was in the same size range as that of the administered AgNPs. No silver granules were detected by TEM in the liver.ConclusionsThe results of the present study demonstrate that the organ distribution of silver was similar when AgNPs or AgAc were administered orally to rats. The presence of silver granules containing selenium and sulfur in the intestinal wall of rats exposed to either of the silver forms suggests a common mechanism of their formation. Additional studies however, are needed to gain further insight into the underlying mechanisms of the granule formation, and to clarify whether AgNPs dissolve in the gastrointestinal system and/or become absorbed and translocate as intact nanoparticles to organs and tissues.

Carbon-enhanced inductively coupled plasma mass spectrometric detection of arsenic and selenium and its application to arsenic speciation

Addition of carbon as methanol or ammonium carbonate to the aqueous analyte solutions in combination with increased plasma power input enhanced the inductively coupled plasma mass spectrometry (ICP-MS) signal intensities of arsenic and selenium. In the presence of the optimum 3% v/v methanol concentration the signal intensities achieved were about 4500–5000 counts s–1 per ng ml–1 of arsenic and about 700–1100 counts s–1 per ng ml–1 of selenium (82Se), corresponding to enhancement factors of 3.5–4.5 compared with aqueous solution for the two elements. Differences in sensitivity (calculated on the basis of analyte atom) were observed between the individual arsenic species and between the selenium species in aqueous as well as in carbon-added solutions. The presence of 3% v/v methanol in the analyte solutions doubled the level of the background signal for arsenic and selenium, but its fluctuation (noise) was not increased. Therefore, the observed increase in analyte sensitivity led to a similar increase in signal-to-noise ratio. The addition of carbon as ammonium carbonate enhanced the arsenic signal by a similar factor but caused severe contamination of the ICP-MS instrument by carbon. In the 3% v/v methanol solutions of arsenic and selenium the signal intensity from antimony (internal standard) was enhanced by a factor of 1.5, which indicates that the enhancement effect of the arsenic and selenium signals by methanol is only to a limited extent caused by improved sample transport/nebulization efficiency. It is proposed that an increased population of carbon ions or carbon-containing ions in the plasma facilitates a more complete ionization of analytes lower in ionization energy than carbon itself. The enhanced detection power for arsenic was applied to arsenic speciation by high-performance liquid chromatography (HPLC)–ICP-MS, and made possible the detection of the arsenocholine ion (AsC) in extracts of shrimp at the 5–10 ng g–1 concentration level. The limit of detection was improved by a factor of 3.4 after addition of methanol and was 4.7 ng g–1 as the AsC ion.

Monitoring the content and intake of trace elements from food in Denmark

The content of cadmium, lead, nickel, mercury and selenium in 83 foods was monitored from 1993 to 1997. In comparison with similar results from 1988 to 1992, a general decrease in lead levels had occurred, whereas the contents of cadmium, nickel, mercury and selenium were stable or declined only slightly. The distribution in dietary intake of the five trace elements was estimated by combining the mean trace element concentrations with food consumption data from 1837 Danes aged 15–80 years. The lead intake for 1993–97 showed a decrease in comparison with similar estimates from the previous monitoring cycles: 1983–87 and 1988–92. The intake of cadmium and mercury decreased to a lesser extent, whereas the intake of selenium and nickel remained unchanged in the same period. The dietary intake of trace elements was compared with the provisional tolerable weekly intake (PTWI). The 95th percentile of the distribution in cadmium intake amounts to 34% of PTWI, which is relatively high, and therefore calls for a more detailed future risk assessment. The intakes of lead and mercury were 11% of PTWI and, like the intake of nickel, did not cause any health concern in the adult population. The Danes ingest close to 100% of the Nordic Nutrition Recommendation for selenium at 50 μg day-1, and no individuals had an intake less than the lower limit of 20 μg day-1.

Arsenic speciation in seafood samples with emphasis on minor constituents: an investigation using high-performance liquid chromatography with detection by inductively coupled plasma mass spectrometry

Extracts of 11 samples of shrimp, crab, fish, fish liver, shellfish and lobster digestive gland (hepatopancreas), including five certified reference materials, were investigated for their contents of arsenic compounds (arsenic speciation). The cation-exchange high-performance liquid chromatography procedure was optimized to separate six cationic arsenicals present in the samples with internal chromatographic standardization by the trimethylselenonium ion, which was detected at m/z 82 (82Se), in addition to arsenic at m/z 75, by inductively coupled plasma mass spectrometry. The content of each species (as arsenic atom) relative to the total arsenic extracted from the samples were: arsenobetaine 19–98%, arsenocholine and trimethylarsine oxide 0–0.6% and the tetramethylarsonium ion 0–2.2%. Additionally, an unknown arsenic species (U1) was present at 3.1–18% in the shellfish and in the lobster digestive gland, and another unknown (U2) was present at 0.2–6.4% in all samples. The contents of arsenite and arsenate were 0–1.4%, dimethylarsinate 8.2–29% while monomethylarsonate was detected only in oyster at 0.3% of the total extracted arsenic. Finding tetramethylarsonium ion and arsenocholine in a variety of samples indicates steps of a biosynthetic pathway of arsenic leading to arsenobetaine in the marine environment. The intake of inorganic arsenic via ingestion of the seafood samples that were analysed did not represent a toxicological problem to humans. The limits of detection (LOD) were in the range 10–50 ng g–1(dry mass) with the exception of arsenobetaine for which the LOD was 360 ng g–1.

Effects of prenatal exposure to surface-coated nanosized titanium dioxide (UV-Titan). A study in mice

BackgroundEngineered nanoparticles are smaller than 100 nm and designed to improve or achieve new physico-chemical properties. Consequently, also toxicological properties may change compared to the parent compound. We examined developmental and neurobehavioral effects following maternal exposure to a nanoparticulate UV-filter (UV-titan L181).MethodsTime-mated mice (C57BL/6BomTac) were exposed by inhalation 1h/day to 42 mg/m3 aerosolized powder (1.7·106 n/cm3; peak-size: 97 nm) on gestation days 8-18. Endpoints included: maternal lung inflammation; gestational and litter parameters; offspring neurofunction and fertility. Physicochemical particle properties were determined to provide information on specific exposure and deposition.ResultsParticles consisted of mainly elongated rutile titanium dioxide (TiO2) with an average crystallite size of 21 nm, modified with Al, Si and Zr, and coated with polyalcohols. In exposed adult mice, 38 mg Ti/kg was detected in the lungs on day 5 and differential cell counts of bronchoalveolar lavage fluid revealed lung inflammation 5 and 26-27 days following exposure termination, relative to control mice. As young adults, prenatally exposed offspring tended to avoid the central zone of the open field and exposed female offspring displayed enhanced prepulse inhibition. Cognitive function was unaffected (Morris water maze test).ConclusionInhalation exposure to nano-sized UV Titan dusts induced long term lung inflammation in time-mated adult female mice. Gestationally exposed offspring displayed moderate neurobehavioral alterations. The results are discussed in the light of the observed particle size distribution in the exposure atmosphere and the potential pathways by which nanoparticles may impart changes in fetal development.

Quantitative Characterization of Gold Nanoparticles by Field-Flow Fractionation Coupled Online with Light Scattering Detection and Inductively Coupled Plasma Mass Spectrometry

An analytical platform coupling asymmetric flow field-flow fractionation (AF(4)) with multiangle light scattering (MALS), dynamic light scattering (DLS), and inductively coupled plasma mass spectrometry (ICPMS) was established and used for separation and quantitative determination of size and mass concentration of nanoparticles (NPs) in aqueous suspension. Mixtures of three polystyrene (PS) NPs between 20 and 100 nm in diameter and mixtures of three gold (Au) NPs between 10 and 60 nm in diameter were separated by AF(4). The geometric diameters of the separated PS NPs and the hydrodynamic diameters of the Au and PS NPs were determined online by MALS and DLS, respectively. The three separated Au NPs were quantified by ICPMS and recovered at 50-95% of the injected masses, which ranged between approximately 8-80 ng of each nanoparticle size. Au NPs adhering to the membrane in the separation channel was found to be a major cause for incomplete recoveries. The lower limit of detection (LOD) ranged between 0.02 ng Au and 0.4 ng Au, with increasing LOD by increasing nanoparticle diameter. The analytical platform was applied to characterization of Au NPs in livers of rats, which were dosed with 10 nm, 60 nm, or a mixture of 10 and 60 nm nanoparticles by intravenous injection. The homogenized livers were solubilized in tetramethylammonium hydroxide (TMAH), and the recovery of Au NPs from the livers amounted to 86-123% of their total Au content. In spite of successful stabilization with bovine serum albumin even in alkaline medium, separation of the Au NPs by AF(4) was not possible due to association with undissolved remains of the alkali-treated liver tissues as demonstrated by electron microscopy images.

Bioavailability and speciation of arsenic in carrots grown in contaminated soil

Carrots were grown in seven experimental plots (A-G) containing mixtures of arsenic-contaminated and uncontaminated soil at concentrations ranging from 6.5 to 917 microgram g(-1) (dry mass). The carrots harvested from plots A-D (6.5-338 microgram g(-1) arsenic in the soil mixtures) showed a gradually increasing depression of growth with increasing level of contamination. At the experimental plots E-G with soil arsenic concentrations above 400 microgram g(-1) no carrots developed. Whether this effect was caused by arsenic or the concomitant copper content which ranged from 11 to 810 microgram g(-1) in the soil mixtures is unknown. The arsenic species extracted from the soils and carrots were separated and detected using anion-exchange HPLC coupled with ICP-MS. In the less contaminated soils from plots A and B arsenite (AsIII) was more abundant than arsenate (Asv) in the soil using 1 mmole 1-1 calcium nitrate as extractant. In the soils from plots C and D however, Asv dominated over AsIII whereas in the corresponding carrots Asv and AsIII were found at similar concentrations. Methylated arsenic species were sought after but not detected in any of the samples. The soil-to-carrot uptake rate (bioavailability) of arsenic was 0.47 +/- 0.06% (average +/- one standard deviation) of the arsenic content in the soils from plots A-D. In contrast to arsenic, the increasing copper content in the soils from plot A through D was not available to the carrots as the concentration of this element did not increase with increasing soil copper content. The ingestion of the potentially toxic inorganic arsenic via consumption of carrots grown in soil contaminated at 30 microgram g(-1) in arsenic (plot B) was conservatively estimated at 37 microgram week (-1). This was equivalent to only 4% of the provisional tolerable weekly intake (PTWI) for inorganic arsenic as suggested by the WHO and was therefore toxicologically safe. Consumption of carrots grown in more intensely arsenic-contaminated soils, however, would lead to a higher intake of inorganic arsenic and is therefore not recommended.

Arsenic speciation by liquid chromatography coupled with ionspray tandem mass spectrometry

Ionspray mass spectrometry, a well established organic analysis technique, has been coupled to high-performance liquid chromatography for speciation of organic arsenic compounds. The ionspray source and differentially pumped interface of the mass spectrometer were operated in dual modes for elemental and molecular analysis. Tandem mass spectrometry was employed to increase selectivity. Dual mode detection was applied to demonstrate the utility of the technique for analysis of nine organoarsenic standards, including four dimethylarsinylriboside derivatives (arsenosugars). Structural fragmentation patterns showing molecular dissociation through an expected common product ion were obtained for the four arsenosugars. Molecular mode detection was utilized for qualitative verification of speciation analysis by high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry of organoarsenicals in plaice and for analysis of an arsenosugar in extracts of oyster and mussel.

Extracellular thiol-assisted selenium uptake dependent on the xc− cystine transporter explains the cancer-specific cytotoxicity of selenite

The selenium salt selenite (SeO32−) is cytotoxic in low to moderate concentrations, with a remarkable specificity for cancer cells resistant to conventional chemotherapy. Our data show that selenium uptake and accumulation, rather than intracellular events, are crucial to the specific selenite cytotoxicity observed in resistant cancer cells. We show that selenium uptake depends on extracellular reduction, and that the extracellular environment is a key factor specific to selenite cytotoxicity. The extracellular reduction is mediated by cysteine, and the efficacy is determined by the uptake of cystine by the xc− antiporter and secretion of cysteine by multidrug resistance proteins, both of which are frequently overexpressed by resistant cancer cells. This mechanism provides molecular evidence for the existence of an inverse relationship between resistance to conventional chemotherapy and sensitivity to selenite cytotoxicity, and highlights the great therapeutic potential in treating multidrug-resistant cancer.

Speciation of eight arsenic compounds in human urine by high-performance liquid chromatography with inductively coupled plasma mass spectrometric detection using antimonate for internal chromatographic standardization

Four anionic and four cationic arsenic compounds in urine were separated by anion- and cation-exchange high-performance liquid chromatography and detected by inductively coupled plasma mass spectrometry (ICP-MS) at m/z 75. The species were the anions arsenite, arsenate, monomethylarsonate and dimethylarsinate and the cations arsenobetaine, trimethylarsine oxide, arsenocholine and the tetramethylarsonium ion. Hexahydroxyantimonate(III) was co-chromatographed with the arsenic anions but detected at m/z 121 and used as an internal standard for their qualitative analysis. Arsenite was prone to oxidation to arsenate in urine but was stable after at least 4-fold dilution of the urine with water. Arsenite was unstable in both urine samples and standard mixtures when diluted with the basic (pH 10.3) mobile phase used for anion chromatography. This could not be prevented by adding ascorbic acid as antioxidant. The argon chloride interference at m/z 75 was eliminated by chromatographic separation of the chloride present in the sample from the arsenic analytes. The ClO+ ion detected at m/z 51 and 53 was used to monitor the retention time of chloride in the anion-exchange system. The chloride eluted about 100 s after the last analyte peak and the intensity of ArCl+ was negligible even after injection of a 1% NaCl solution (less than 200 ions s–1). The recovery of all arsenic species in urine was close to 100%. The chromatographic peaks were evaluated by their peak heights and calibration was carried out by the method of standard additions. The calibration graphs were linear for all species (r > 0.999). The limits of detection were 3–6 ng cm–3 for the cations and 7–10 ng cm–3 for the anions in urine.