Determination of trace amounts of selenium in poultry feedstuffs by gas chromatography.

In view of the importance of establishing reliable selenium concentration levels in different kinds of feedstuffs, the purpose of this work was to develop optimum experimental conditions for the isolation and GC determination of selenium as its chelate with 4-nitro-1,2-diaminobenzene. It was shown that ignition of the sample in an oxygen flask followed by reduction of Se(VI) to Se(IV) and the formation of 5-nitro-2,1,3-benzoselenadiazole chelate in HCl medium is a relatively rapid procedure giving a low blank value and allowing the determination of selenium in commercial feedstuffs and similar biological samples. The method was validated by the analysis of suitable certified or standard reference materials.

Preliminary studies on arsenic species in some environmental samples.

Arsenic compounds have been determined in some environmental samples from the German Environmental Specimen Bank (ESB) (marine mussels, freshwater mussel and fish, sea-gull eggs) and certified reference materials (DORM-1, DOLT-1, NBS Oyster Tissue) after separation by open column cation and anion exchange chromatography by two different methods of total arsenic determination in separated fractions (instrumental neutron activation analysis or hydride generation atomic absorption spectrometry). Arsenobetaine has been identified as the major species in all the different materials.

Determination of uranium at trace levels by radiochemical neutron-activation analysis employing radioisotopic yield evaluation.

Nanogram and picogram quantities of uranium were determined in biological materials by radiochemical neutron-activation analysis. Two different approaches using either (239)U or (239)Np were employed for cross-checking, and the question of negative errors due to incomplete acid dissolution of any possible inorganic (siliceous) fraction was studied. In the first and main approach, radiochemical separation of the short-lived (239)U (23.5 min) nuclide was based on TBP extraction following rapid conventional wet-ashing. Addition of large amounts of uranium carrier (ca. 50 mg) allowed the chemical yield to be evaluated from the gamma spectrum of the isolated fraction by means of the 186 keV peak of (235)U. In the second approach, the longer-lived (239)Np (56.5 hr) daughter was separated by anion-exchange; this nuclide allowed use of lengthier dissolution procedures employing total decomposition with hydrofluoric acid. Nanogram quantities of (237)Np were irradiated simultaneously with the sample and an aliquot of the resulting solution containing (237)Np and (238)Np (51 hr) was added prior to sample destruction, these isotopes serving as carrier and yield tracer, respectively. Results are presented for a series of reference materials. The methodologies and results from the two approaches are discussed and evaluated.

Activation analysis for mercury in biological samples at nanogram level.

A new method has been devised for determining mercury in samples of biological origin. It is based on complete ignition of the sample in a silica tube, trapping volatile interfering activities such as bromine or chlorine, and selectively adsorbing mercury on a strip of filter paper which has been previously impregnated with elemental selenium. This strip is later counted for quantitative evaluation. The versatility of the method has been demonstrated by the analysis of a wide range of samples such as water, cellulose, flour, fish solubles or animal blood samples with mercury contents between 1 and 200 ng g of sample.

Simultaneous neutron-activation determination of selenium and mercury in biological samples by volatilization.

A method is described for the determination of selenium together with mercury in biological samples by neutron-activation analysis based on quantitative volatilization of both elements. The technique originally developed for mercury, based on pyrolysis with filtration of undesirable impurities and selective trapping from the gas phase, is now extended to selenium. The radionuclides (197)Hg and (75)Se, from one sample, are trapped separately and counted in a well-type NaI(Tl) detector and gamma-spectrometer for maximum sensitivity. The method has been tested by comparative analyses and analyses of standard biological materials, and gives good results. It is simple and is especially effective in studies of the interaction of mercury and selenium in biological systems; a positive correlation for these elements was found for human tissues. On décrit une méthode pour le dosage du sélénium conjointement au mercure dans les échantillons biologiques par analyse par activation de neutrons basée sur la volatilisation quantitative des deux éléments. La techniqu initialement développée pour le mercure, basée sur la pyrolyse avec filtration des impuretés indésirables et captage sélectif de la phase gazeuse, est maintenant étendue au sélénium. Les radionuclides (197)Hg et (75)Se, d'un échantillon, sont captés séparément dans un détecteur NaI(Tl) du type puits et un spectromètre gamma pour la sensibilité maximale. La méthode a été essayée par des analyses comparatives et des analyses de produits biologiques étalons, et donne de bons résultats. Elle est simple et particulièrement efficace dans les études de l'interaction du mercure et du sélénium dans des systèmes biologiques; on a trouvé une corrélation positive pour ces éléments pour des tissus humains.

A modified method for the determination of methylmercury by gas chromatography.

A simple modification of the Westöö extraction procedure for methylmercury and its determination by gas chromatography (GC) is presented. The cysteine clean-up step has been modified, with use of cysteine-impregnated paper instead of cysteine solution. Methylmercury bromide is extracted from the sample into toluene and is selectively adsorbed on the cysteine paper. Interfering compounds are washed from the paper with toluene. The isolated methylmercury is set free with sulphuric acid containing bromide, extracted into benzene and determined by GC. The modification of the extraction procedure results in good recovery and reproducibility for various biological and environmental samples, good sensitivity with a detection limit of 0.1 ng/g, avoidance of difficulties arising from emulsion formation, cleaner chromatograms, and faster analysis. It is particularly suitable for determination of low levels of MeHg.

Determination of traces of indium, manganese, arsenic and antimony in zinc by neutron-activation analysis.

Destructive activation determination of the trace elements indium, manganese, arsenic and antimony in different samples of pure zinc metal by solvent extraction techniques is described. Determination of indium and manganese is based on the quantitative co-precipitation of both elements with lanthanum hydroxide, followed by their extraction with sodium diethyldithiocarbamate in the presence of potassium cyanide and their subsequent separation by selective stripping. The quantitative determination of arsenic and antimony is based on the extraction of their iodides from sulphuric acid solution with toluene.

Determination of arsenic compounds in reference materials by HPLC-(UV)-HG-AFS.

Arsenic compounds were determined in six reference materials of biological origin. None of them has yet been certified for arsenic compounds but some are in the process of certification; for most of these reference materials indicative literature values are available. Eight commonly used arsenic standards were used for quantification using a recently developed hyphenated speciation system comprising high performance liquid chromatography (HPLC) and atomic fluorescence spectrometry (AFS), interfaced via a UV-photoreactor and a hydride generation (HG) unit. Absolute detection limits were ca. 0.2 and 0.4 ng As for separation on anion and cation exchange columns, respectively. Our results agree well with indicative literature values which were generated by different authors using various separation and detection methods. The HPLC-(UV)-HG-AFS system validated in this way is suitable for quantification of eight arsenic compounds. Moreover, the system is capable of separation of at least six more compounds in the mentioned reference materials, of which two could be attributed to arsenosugars (OH and phosphodiester form) but due to the lack of standards, quantification was not possible. For accurate and extensive speciation analysis the availability of certified reference materials and standards for arsenic compounds should be promoted.

Uranium content of blood, urine and hair of exposed and non-exposed persons determined by radiochemical neutron activation analysis, with emphasis on quality control.

Recent reviews have highlighted the diversity (and sparsity) of data for uranium concentrations in body fluids, bone and soft tissues, and it is unclear to what extent this reflects genuine geographical or biological variations. In the present work, a very sensitive radiochemical neutron activation analysis technique (detection limit, 1-2pg g-1) was applied to an exploratory study of uranium levels in hair, urine and blood of non-exposed and occupationally exposed persons. Since quality control in much previous work has been somewhat neglected, this aspect was emphasized by the use of a wide range of suitable reference materials, by standard addition experiments, and by care in sampling and sample handling. For controls, typical levels found in hair, urine and blood were 10 ng g-1, 10 and 5 ng l-1, respectively. The urine values correspond to the lower end of the literature data, while for blood our value is about two orders of magnitude lower than the presently quoted level. In the case of occupationally exposed persons, it was found that hair shows some promise as an indicator of exposure.

Vanadium in foods and in human body fluids and tissues.

Using neutron activation analysis, vanadium was analysed in a range of foods, human body fluids and tissues. On the basis of these results and those of other workers, it was concluded that daily dietary intake amounts to some tens of micrograms. Analysis of body fluids (including milk, blood and excreta) and organs and tissues provided an estimate for the total body pool of vanadium in man of about 100 microgram. Vanadium was not detectable in blood and urine at the level of 0.3 ng/g, while low levels were found in muscle, fat, bone, teeth and other tissues. The relationship between dietary intake to pulmonary absorption is discussed in relation to the occurrence of vanadium in man-made air particulates. The very low levels found in milks and eggs suggest minimal vanadium requirements in growth. The findings are discussed in the light of previous results and also in relation to the possible essentiality of vanadium.

On the vanadium and tin contents of diet and human blood.

The vanadium and tin contents of total diet samples collected in five Italian towns are given, and discussed in relation to literature data, their status as essential elements and their levels in human blood. Tin in blood was found to be below 2 ng ml-1 in a group of 14 subjects, and barely detectable even after oral consumption of 60 mg of tin.

Trace elements in some human milk samples by radiochemical neutron activation analysis.

Some trace element contents of samples of human milk, mainly colostrum and transitional milk, collected in the Ljubljana area of Yugoslavia, are reported. Analyses of As, Cd, Co, Cu, Hg, I, Mn, Sb, Se, Sn, V and Zn were performed by radiochemical neutron activation analysis, and the results are discussed briefly in the light of literature values.

Trace element concentrations in higher fungi.

The concentrations of ten trace elements, As. Br, Cd, Cu, Hg I, Mn, Se, Zn and V, have been determined in up to 27 species of higher fungi from several sites in Slovenia, Yugoslavia. Analyses were based on destructive neutron activation techniques. Data are presented and compared with the concentrations found in soils. Previously values were non-existent or scanty for these elements, so that the data represent typical levels for basidiomycetes. In addition to confirming high levels of mercury in many species, the survey also found that cadmium is accumulated to a surprising extent by most fungi, the average value being 5 ppm. Among other accumulations found was bromine by the genus Amanita, and selenium by the edible Boletus. Correlation analysis between all pairs of trace elements gave values for r of from 0.75 to 0.43 for 7 pairs (Cu and Hg, 0.75; Se and As, 0.69). As well as these features of biochemical interest, the values found and the pattern of accumulation suggest potential uses of fungi in environmental studies.

Prompt and delayed NAA techniques for the characterization of specimen bank materials.

The combined application of instrumental-, radiochemical- and prompt gamma neutron activation analysis to two spruce shoot materials from the German Environmental Specimen Bank (ESB) resulted in information on 50 elements, covering more than 50% of the total mass. Comparison of the element concentrations in the 'fingerprint' mode clearly indicated a different status of heavy metal pollution at the two distinct collecting sites.

Vanadium levels in hair and blood of normal and exposed persons.

Vanadium was determined by both instrumental neutron activation analysis (INAA) and NAA with radiochemical separation (RNAA) in hair of normal children and of children potentially exposed by accidental drinking of vanadium contaminated water (long-term, low-dose exposure). Vanadium hair levels in the two groups did not differ significantly and were in the range 46-313 micrograms/kg (median 98 micrograms/kg) and 24-235 micrograms/kg (median 88 micrograms/kg for the normal and exposed groups, respectively. Using RNAA with proven reliability at the ultratrace level, vanadium was determined in whole blood of the exposed and normal children, normal adults and workers professionally exposed to vanadium in a factory producing vanadium pentoxide. Significantly increased vanadium concentrations were found in blood of exposed children (range 0.018-0.239 micrograms/l, median 0.078 micrograms/l) compared to normal children (range 0.024-0.226 micrograms/l, median 0.042 micrograms/l), while no differences could be detected between blood vanadium levels of normal children and normal adults (range 0.032-0.095 micrograms/l, median 0.056 micrograms/l). Preliminary results for vanadium in blood of occupationally highly exposed persons showed values 2-4 orders of magnitude higher than in normal adults.

Some improvements in the quality of NAA procedures and the reliability of the results.

After considering the need for quality control in NAA, the concept of quality in NAA procedures themselves is discussed, and some important factors identified. Two approaches to improve quality are then described in more detail. The first concerns the unique ability of NAA using different isotopic reactions and different modes (INAA/RNAA) to provide independent data sets in the same laboratory, thus allowing internal validation or crosschecking. The second discusses the need for chemical yield measurements in RNAA and the advantages of the radioisotopic tracer technique. Some recent advances and further possibilities for this use of tracers are listed.

Vanadium determination at the ultra-trace level in biological reference materials and serum by radiochemical neutron activation analysis.

In order to help resolve present inconsistencies of two orders of magnitude or more in reported levels of vanadium in human serum and blood, a totally postirradiation radiochemical neutron activation analysis (NAA) method was further developed and applied to some pertinent nanogram and subnanogram reference materials. In particular, the second generation human serum reference material of Versieck was found to contain a value of 0.67 +/- 0.05 ng/g dry wt., corresponding to 0.061 +/- 0.005 /4/ ng/mL original fresh serum. Results are also reported for some other appropriate CRMs. Additionally, a small-scale study in 10 normal subjects (5 m, 5 f) revealed levels similar to those in the serum reference material and in agreement with the lowest data reported in the literature. Discussion of pitfalls of vanadium determination and the use of reference materials is included.

Comprehensive RNAA of cadmium, cobalt, nickel, and copper using 109Cd, 57Co, and reactor-produced 67Cu as radioisotopic yield monitors.

An existing radiochemical NAA procedure for Cd, Co, and Cu was improved to allow determination of individual radiochemical yields by the radioisotopic tracer technique, thus eliminating errors owing to variable recovery. 109Cd was used as tracer for Cd determination via 115Cd/115mIn, 57Co for Co via 60Co, and potentially for Ni via 58Co, whereas as a novelty 67Cu, produced by reactor irradiation of ZnO of natural isotopic composition (by the 67Zn [n,p] 67Cu reaction) was used for Cu via the indicator nuclide 64Cu. The simple production and purification of 67Cu by anion exchange is described. Results for biological RMs are given and discussed.

Preliminary study on the determination of selenium compounds in some selenium-accumulating mushrooms.

Using various chromatographic techniques (size exclusion, anion exchange, and cation exchange) combined with several detectors (neutron activation analysis and atomic fluorescence spectrometry), an attempt was made to characterize selenium compounds in some edible, selenium-accumulating mushrooms (Albatrellus pes-caprae and Boletus edulis). The mushrooms contained mostly low-molecular-weight (6 kDa) selenium compounds. After proteolysis, only a small fraction of the extractable selenium could be identified as selenite (3.0-9.2%, Albatrellus pes-caprae), selenocystine (minor, Albatrellus pes-caprae; 7.5%, Boletus edulis), or selenomethionine (1.0%, Boletus edulis), leaving the form of the bulk still to be elucidated.

Minor and trace elements in human milk from Guatemala, Hungary, Nigeria, Philippines, Sweden, and Zaire. Results from a WHO/IAEA joint project.

Concentrations of As, Ca, Cd, Cl, Co, Cr, Cu, F, Fe, Hg, I, K, Mg, Mn, Mo, Na, Ni, P, Pb, Sb, Se, Sn, V, and Zn were determined in human whole milk samples from Guatemala, Hungary, Nigeria, Philippines, Sweden, and Zaire; in most of these countries, three groups of subjects representing different socioeconomic conditions were studied. Analytical quality control was a primary consideration throughout. The analytical techniques used were atomic absorption spectrophotometry, atomic emission spectrometry with an inductively coupled plasma, colorimetry, electrochemistry, using an ion-selective electrode and neutron activation analysis. The differences between median concentrations of Ca, Cl, Mg, K, Na, and P (minor elements) were lower than 20% among the six countries. Among trace elements, concentrations observed in Filipino milk for As, Cd, Co, Cr, Cu, F, Fe, Mn, Mo, Ni, Pb, Sb, Se, and V were higher than for milk samples from other countries. The remaining five countries showed a mixed picture of high and low values. In the case of at least some elements, such as, F, I, Hg, Mn, Pb, and Se, the environment appears to play a major role in determining their concentrations in human milk. The nutritional status of the mother, as reflected by her socioeconomic status, does not appear to influence significantly the breast milk concentrations of minor and trace elements. Significant differences exist between the actual daily intakes observed in this study and current dietary recommendations made by, for example, WHO and the US National Academy of Sciences. These differences are particularly large (an order of magnitude or more!) for Cr, F, Fe, Mn, and Mo; for other elements, such as, Ca, Cu, Mg, P, and Zn, they amount to at least a factor 2. In the opinion of the present authors, these findings point to the need for a possible reassessment of the dietary requirements of young infants with respect to minor and trace elements, particularly for the elements Ca, Cr, Cu, F, Fe, Mg, Mn, Mo, P, and Zn.