Elemental hair analysis: A review of procedures and applications.

Although exogenous contamination and unreliable reference values have limited the utility of scalp hair as a biomarker of chemical elements exposure, its use in toxicological, clinical, environmental and forensic investigations is growing and becoming more extensive. Therefore, hair elemental analysis is reviewed in the current manuscript which spans articles published in the last 10 years. It starts with a general discussion of history, morphology and possible techniques for elemental analysis, where inductively coupled plasma-mass spectrometry (ICP-MS) is clearly highlighted since this technique is leading quantitative ultra-trace elemental analysis. Emphasis over sampling, quality assurance, washing procedures and sample decomposition is given with detailed protocols compiled in tables as well as the utility of hair to identify human gender, age, diseases, healthy conditions, nutrition status and contamination sites. Isotope ratio information, chemical speciation analysis and analyte preconcentration are also considered for hair. Finally, the potential of laser ablation ICP-MS (LA-ICP-MS) to provide spatial resolution and time-track the monitoring of elements in hair strands instead of conventional bulk analysis is spotlighted as a real future trend in the field.

Hypoxia acclimation protects against oxidative damage and changes in prolactin and somatolactin expression in silver catfish (Rhamdia quelen) exposed to manganese.

The aim of this study was to assess the Mn toxicity to silver catfish considering Mn accumulation and oxidative status in different tissues, as well as pituitary hormone expression after acclimation to hypoxia. Silver catfish acclimated to hypoxia for 10 days and successively exposed to Mn (9.8 mg L(-1)) for an additional 10 days exhibited lower Mn accumulation in plasma, liver, kidneys and brain and prevented the hematocrit decrease observed in the normoxia group. Hypoxia acclimation also modified Mn-induced oxidative damage, which was observed by lower reactive species (RS) generation in gills and kidneys, decreased lipid peroxidation (LP) levels in gills, liver and kidneys and decreased protein carbonyl (PC) levels in liver, kidneys and brain. Manganese accumulation showed positive correlations with LP levels in gills and kidneys, as well as with PC levels in gills, liver and brain. In addition, hypoxia acclimation and Mn exposure increased catalase (CAT) activity in gills and kidneys and Na(+)/K(+)-ATPase activity in gills, liver and brain. Silver catfish that were acclimated under normoxia and exposed to Mn displayed increased pituitary prolactin (PRL) and decreased somatolactin (SL) expression. Interestingly, hypoxia acclimation prevented hormonal fluctuation of PRL and SL in fish exposed to Mn. These findings indicate that while the exposure of silver catfish to Mn under normoxia was related to metal accumulation and oxidative damage in tissues together with endocrine axis disruption, as represented by PRL and SL, hypoxia acclimation reduced waterborne Mn uptake, thereby minimizing oxidative damage and changes in hormonal profile. We hypothesized that moderate hypoxia is able to generate adaptive responses, which may be related to hormesis, thereby ameliorating Mn toxicity to silver catfish.

Moderate hypoxia is able to minimize the manganese-induced toxicity in tissues of silver catfish (Rhamdia quelen).

The aim of this study was to compare the effects of manganese (Mn) on silver catfish exposed to different levels of dissolved oxygen. Silver catfish (Rhamdia quelen) were exposed to increasing concentrations of Mn (4.2, 8.4 or 16.2mgL(-1)) under either normoxia (100 percent saturation) or moderate hypoxia (51.87 percent saturation) for 15 days. Under normoxia, Mn exposure increased lipid peroxidation (LP) in brain and kidney; it increased gluthatione (GSH) levels in brain and decreased catalase (CAT) activity in both tissues. Moderate hypoxia was able to prevent Mn-induced LP in brain and to reduce this oxidative parameter in kidney; GSH level was increased in brain, while CAT activity was reduced in both tissues. Activity of isolated mitochondria of liver and gills was reduced by Mn exposure under both levels of dissolved oxygen, but this effect was more prominent in normoxia. As expected, liver, kidney and gills showed an increase of Mn accumulation according to waterborne levels, and these parameters presented positive relationship. The highest waterborne Mn (8.4 and 16.2mgL(-1)) resulted in greater accumulation under normoxia, indicating that moderate hypoxia can stimulate mechanisms capable of reducing Mn accumulation in tissues (though not in blood). Moderate hypoxia can be considered a stress factor and Mn an aquatic anthropogenic contaminant. Therefore we hypothesized that these two conditions together are able to invoke defense mechanisms in juvenile silver catfish, acting in a compensatory form, which may be related to adaptation and/or hormesis.

Effect of wheat bran and flaxseed on cadmium effects and retention in rats.

Dietary fiber can affect cadmium (Cd) absorption and toxicity, but the effect appears to depend on the type of dietary fiber. The aim of the present study was to compare the effect of dietary sources containing distinct amounts of soluble and insoluble fiber on Cd absorption, accumulation and toxicity in growing rats. The absorption of essential macrominerals (Ca, P and Mg) was also evaluated. Animals received a nutritionally balanced diet with cellulose (cel - control), wheat bran or flaxseed as the fiber source with 0 or 50 mg Cd kg(-1) diet, during 30 days. Cd exposure reduced body weight gain, feed efficiency ratio, epididymal fat relative weight and liver relative weight, and increased plasma alanine aminotransferase activity in all fiber groups. The apparent Cd absorption was similar among Cd-groups, but the flax-Cd group had a higher hepatic and renal Cd concentration. Cd decreased the absorption of Ca and P, and increased Mg absorption in the wheat bran and flaxseed groups, but not in the cel group. Although the different fiber sources investigated had no effect on Cd toxicity, the major soluble fiber source, flaxseed, increased Cd retention. Thus, caution should be taken in the intake of flaxseed by Cd-exposed populations.

Delayed biochemical changes induced by mercury intoxication are prevented by zinc pre-exposure.

This work evaluated the delayed effects of mercury and the effectiveness of zinc in preventing such effects. Pups were pre-treated with 1 daily dose of ZnCl(2) (27 mg/kg/day, by subcutaneous injections) from 3rd to 7th postnatal day and received 1 daily dose of 5 mg/kg of HgCl(2), for 5 subsequent days (8-12 days old). Animals were euthanized 21 days after the end of Hg-exposure. Porphobilinogen-synthase activity as well as zinc and mercury contents was determined in the liver and kidneys. Alanine aminotransferase, aspartate aminotransferase and lactic dehydrogenase activities as well as urea, creatinine and glucose levels were analyzed in plasma or serum. Some animals were considered more sensitive to mercury, since they did not recover the body weight gain and presented an increase of renal and hepatic mercury content, urea and creatinine levels; a decrease in renal porphobilinogen-synthase and alanine aminotransferase activities, as well as a decrease in the liver and an increase in kidney weights. Some animals were considered less sensitive to mercury because they recovered the body weight and presented no biochemical alterations in spite of mercury in the tissues. Zinc prevents partially or totally the alterations caused by mercury even those that persisted for a long time after the end of exposure. These findings suggest that there is difference among the animals regarding the sensitivity to mercury.

Antioxidant system activation by mercury in Pfaffia glomerata plantlets.

Oxidative stress caused by mercury (Hg) was investigated in Pfaffia glomerata plantlets grown in nutrient solution using sand as substrate. Thirty-day-old acclimated plants were treated for 9 days with four Hg levels (0, 1, 25 and 50 microM) in the substrate. Parameters such as growth, tissue Hg concentration, toxicity indicators (delta-aminolevulinic acid dehidratase, delta-ALA-D, activity), oxidative damage markers (TBARS, lipid peroxidation, and H(2)O(2) concentration) and enzymatic (superoxide dismutase, SOD, catalase, CAT, and ascorbate peroxidase, APX) and non-enzymatic (non-protein thiols, NPSH, ascorbic acid, AsA, and proline concentration) antioxidants were investigated. Tissue Hg concentration increased with Hg levels. Root and shoot fresh weight and delta-ALA-D activity were significantly decreased at 50 microM Hg, and chlorophyll and carotenoid concentration were not affected. Shoot H(2)O(2) concentration increased curvilinearly with Hg levels, whereas lipid peroxidation increased at 25 and 50 microM Hg, respectively, in roots and shoots. SOD activity showed a straight correlation with H(2)O(2) concentration, whereas CAT activity increased only in shoots at 1 and 50 microM Hg. Shoot APX activity was either decreased at 1 microM Hg or increased at 50 lM Hg. Conversely, root APX activity was only increased at 1 microM Hg. In general, AsA, NPSH and proline concentrations increased upon addition of Hg, with the exception of proline in roots, which decreased. These changes in enzymatic and non-enzymatic antioxidants had a significant protective effect on P. glomerata plantlets under mild Hg-stressed conditions.

Influence of cereal bran supplement on cadmium effects in growing rats.

Strategies to diminish cadmium (Cd) absorption are highly desirable especially where Cd exposure due to environmental contamination is still inevitable. Cd toxicity may be influenced by dietary components, such as fiber and minerals. Multimixtures are low-cost cereal bran supplements used in Brazil and in other countries to counteract malnutrition in low-income populations. This study was aimed at evaluating whether multimixture would reduce Cd effects in young rats. Animals received a diet with or without the multimixture plus 0, 5, or 25 mg Cd/kg (control, Cd-5, and Cd-25 groups) during 30 days. The Cd-5 groups were similar to control groups in all parameters analyzed, except in the higher renal Cd concentration. However, the Cd-25 groups had lower biological growth parameters and renal delta-aminolevulinate dehydratase activity, besides higher renal Cd concentration and plasma alanine aminotransferase activity compared to the controls. The multimixture did not prevent Cd effects in the Cd-25 group, but caused a small reduction in renal Cd concentration in the Cd-5 group. Although this multimixture was ineffective to prevent Cd effects at the higher concentration, it seemed to reduce Cd accumulation at the lower Cd dietary concentration, which is similar to levels of human exposure in some polluted areas.

ZnCl2 exposure protects against behavioral and acetylcholinesterase changes induced by HgCl2.

This study examined the effects of inorganic mercury exposure on behavioral and biochemical parameters and investigated the possible preventive effects of zinc on the alterations induced by mercury. Pups were exposed from 3rd to 7th postnatal day to ZnCl2 (27 mg/kg/day, s.c.) and subsequently to HgCl2 (5 doses of 5 mg/kg/day, s.c.). Each litter contained two rats for each treatment. The rats were submitted to behavioral task and litters were killed at 13 or 33 days old for acetylcholinesterase activity assays and for the determination of metal levels. Based on the results obtained from 13-day-old rats, they were divided in two groups of litters that were defined at the end of the experimental period (33 days) as less sensitive rats to mercury and more sensitive rats to mercury in accordance with the recovery of body weight until day 33. The mercury exposure caused accumulation of this metal in cerebrum and cerebellum in all mercury treated rats, and inhibited the cerebellum acetylcholinesterase activity from 13-day-old rats. Besides, the mercury-animals of the most sensitive litters to mercury presented impairment in motor function and muscular strength verified in the beaker test, as well as a reduction of the locomotor and exploratory activities in the open field task. Zinc partially prevented all the alterations induced by mercury exposure and reduced the mercury level accumulated in cerebrum and cerebellum. This study confirms the preventive effect of zinc on behavioral alterations induced by mercury in young rats and demonstrates that the mercury behavioral effects are present even for a long time after the end of the exposure.

Avaliação funcional e histológica da tireoide de ovinos suplementados com fluoreto de sódio por um período de 150 dias / Functional and histological evaluations of thyroid of sheep submitted to sodium fluoride administration

Avaliou-se o efeito da administração de fluoreto de sódio sobre a tireoide de 12 ovinos, distribuídos em dois grupos: o grupo controle (n=6) recebeu somente sal iodado (5g de NaCl/animal + 0,2ppm I/kg MS) e o grupo tratado (n=6), sal iodado (5g de NaCl/animal + 0,2ppm I/kg MS) adicionado de fluoreto de sódio (4,7mg F/kg de peso corporal), durante 150 dias. Amostras de sangue foram coletadas para análise sérica de I e F, triiodotironina (T3) e tetraiodotironina (T4) nos dias zero, 60, 90 e 150 de tratamento. Também se coletou urina, correspondente às 24 horas, para análise da excreção urinária de I e F. Após a eutanásia dos animais, a tireoide foi removida para posterior exame. Quanto ao I urinário, não foi observada diferença estatística entre os grupos controle e tratado e dentro de cada grupo, entre os tempos. As concentrações de T3 e T4 não diferiram entre os grupos e dentro de cada grupo nos diferentes tempos. Quanto à avaliação histopatológica da tireoide, não foram observadas alterações na integridade estrutural. Conclui-se que a administração crônica de fluoreto de sódio não interfere na função e na integridade histológica da glândula tireoide de ovinos com dieta não restritiva de iodo.

The effect of sodium fluoride administration on thyroid function and morphology in sheep was evaluated using 12 lambs. Animals were allotted in two groups: control that received 5g NaCl + 0.2ppm I/kg DM and treated which received the same treatment plus sodium fluoride (4.7mg F/kg BW), daily, for 150 days. Blood samples were collected for determination of F, I, T3, and T4. Urine production was collected for measurement of F and I. After euthanasia at 150 days of treatment, thyroid glands were removed for analysis. No differences were found between or within groups for urinary I. Also, no differences were found for T3 and T4 on serum I between groups or among time points. No histological alterations were found in the thyroids. In conclusion, chronic sodium fluoride treatment did not affect thyroid function and morphology in lambs.

Changes in levels of essential elements in suckling rats exposed to zinc and mercury.

The effect of mercury and its interaction with zinc on the content of essential metals in tissues from neonate rats was investigated. Three-day-old Wistar rats were treated with saline or 27 mg kg(-1)d(-1) ZnCl2 (s.c.) for five consecutive days. From the 8th to the 12th day of life, the rats received one daily dose of saline or 5.0 mg kg(-1) HgCl2 (s.c). Twenty-four hours after the last injection liver, kidneys and blood were collected for metal quantification. The HgCl(2) exposure induced alterations on metal levels, such as increase of Fe, Hg and Zn in liver, decrease of Fe and Mg and increase of Cu and Hg contents in kidneys. The Hg exposure also increased Hg levels in the blood. The treatment with ZnCl2, administered previously to HgCl2, partially prevented the increase of Fe in the liver, and not only prevented the decrease of renal Mg but also increased it to levels higher than those found in control group. The Zn-Hg rats also presented higher renal Cu levels, and showed partially lower blood and hepatic Hg levels and higher renal Hg levels. The pre-administration of Zn caused no severe alterations in levels of essential metals (Cu, Fe, Mg and Mn). In short, Zn appears to be an alternative treatment of Hg poisoning in young animals in comparison to chelating drugs since these have low metal selectivity.

Lead content of dietary calcium supplements available in Brazil.

The lead and calcium content of calcium supplements available in Brazil were determined by graphite furnace and flame atomic absorption spectrometry, respectively. Samples were microwave-digested in concentrated HNO(3). Citric acid was used as a chemical modifier in the lead analysis. Supplements were classified into six categories: oyster industrialized (OI, n=4), oyster prepared in pharmacy (OP, n=3), refined industrialized (RI, n=6), refined prepared in pharmacy (RP, n=3), bone meal (B, n=3), and dolomite (D, n=4). Lead levels (microg g(-1) of measured calcium) were higher in D products (2.33), followed by OI, RP, OP, and RI products (1.46, 1.32, 1.29, 0.75), while B products had levels lower than the limit of quantification (0.02 microg g(-1) unit weight). Daily lead intake of eight supplements exceeded the limit of California, USA (1.5 microg g(-1) calcium), but none exceeded the federal limit of USA (7.5 microg g(-1) calcium) or the provisional tolerable lead intake by FAO/WHO (25 microg kg(-1) per week).

Determination of volatile elements in biological materials by isotopic dilution ETV-ICP-MS after dissolution with tetramethylammonium hydroxide or acid digestion.

Isotopic dilution for the determination of Ag, Cd, Hg, Pb and Tl in biological materials by ETV-ICP-MS is proposed. The sample was simply dissolved with tetramethylammonium hydroxide (TMAH) or acid digested in a microwave furnace, with an on line matrix separation. When the dissolution was employed, Ir was used as a chemical modifier for Hg and Pb and Pd was used for Cd and Tl. No modifier was used for Ag. The pyrolysis temperatures were taken from pyrolysis temperature curves. The on line preconcentration was performed in a flow injection system with solenoid valves and was based on the analyte complexation with ammonium diethyldithiophosphate and sorption of the complexes on C(18) bonded to silica gel in a minicolumn. For the digested sample submitted to the analyte preconcentration procedure, a modifier, Ir, was only used for Hg. For the other analytes, since a low pyrolysis temperature, 300 degrees C, was employed, no modifier was added. The isotopic dilution calibration was applied to two certified materials, bovine liver and dog fish muscle, dissolved with TMAH or acid digested, and to another two certified materials, corn bran and rice flour, acid digested and submitted to analyte preconcentration. The obtained concentration values agree with the certified ones, showing that this calibration procedure leads to accurate results in the determination of low concentrations of volatile elements. Due to simplicity, the dissolution with TMAH is very attractive.

Determination of arsenic(III) and arsenic(V) by electrothermal atomic absorption spectrometry after complexation and sorption on a C-18 bonded silica column.

A flow injection procedure for the separation and pre-concentration of inorganic arsenic based on the complexation with ammonium diethyl dithiophosphate (DDTP) and sorption on a C-18 bonded silica gel minicolumn is proposed. During the sample injection by a time-based fashion, the As(3+)-DDTP complex is stripped from the solution and retained in the column. Arsenic(V) and other ions that do not form complexes are discarded. After reduction to the trivalent state by using potassium iodide plus ascorbic acid, total arsenic is determined by electrothermal atomic absorption spectrometry (ETAAS). Arsenic(V) concentration can be calculated by difference. After processing 6 ml sample volume, the As(3+)-DDTP complexes were eluted directly into the autosampler cup (120 mul). Ethanol was used for column rinsing. Influence of pH, reagent concentration, pre-concentration and elution time and column size were investigated. When 30 mul of eluate plus 10 mul of 0.1% (w/v) Pd(NO(3))(2) were dispensed into the graphite tube, analytical curve in the 0.3-3 mug As l(-1) range was obtained (r=0.9991). The accuracy was checked for arsenic determination in a certified water, spiked tap water and synthetic mixtures of arsenite and arsenate. Good recoveries (97-108%) of spiked samples were found. Results are precise (RSD 7.5 and 6% for 0.5 and 2.5 mug l(-1), n=10) and in agreement with the certified value of reference material at 95% confidence level.

Determination of Mo, U and B in waters by electrothermal vaporization inductively coupled plasma mass spectrometry.

A method for the determination of Mo, U and B in waters by inductively coupled plasma mass spectrometry, using an electrothermal vaporizer for sample introduction, is described. For Mo and U, NH(4)F was chosen as modifier and for B, synthetic sea water plus mannitol were used. The modifier effect was verified and the optimized pyrolysis and vaporization temperatures were obtained from pyrolysis and vaporization curves, together with the transient signals of the analytes. The masses of the modifiers added to the tube were also optimized. The detection limits were 0.018 or 0.30 ng ml(-1) for Mo, 0.03 ng ml(-1) for U and 0.68 ng ml(-1) for B. The analytes were determined in certified waters and the obtained results agree with the certified or recommended values or, in the case of B in sea waters, with the values obtained by other methods. Uranium could not be measured in the sea water samples due to strong memory effect.