Speciation of heavy metals in suspended particulates in urban air†

An anayltical procedure based on a sequential extraction scheme is applied to samples of suspended particulates from an urban area of Barcelona. This area presents high levels of air pollution owing to automotive emissions and industrial activity.

The procedure allows the partitioning of particulate trace metals into four fractions: soluble and exchangeable, bound to carbonates and iron‐manganese oxides, bound to organic matter and residual. In this work, the chemical associations of Fe, Pb, Cu, Mn, Cr and Cd have been investigated.

In the analysed samples, only the cadmium is predominantly in the first fraction whereas the other metals are largely associated to carbonates and Fe‐Mn oxides.     

Analysis of the cyanotoxins anatoxin-a and microcystins in Lesser Flamingo feathers†

Feathers from carcasses of the Lesser Flamingo (Phoeniconaias minor), which had died after ingesting cyanobacterial toxins (cyanotoxins) contained between 0.02 and 30.0?µg microcystin-LR equivalents per gram of feather according to HPLC and ELISA analysis of feather extracts. Anatoxin-a was detected less frequently in the Lesser Flamingo feathers, up to 0.8?µg anatoxin-a per gram of feather being recorded. When feathers from different body regions were analysed and compared for microcystins and anatoxin-a, wing feathers were found to contain the highest concentrations of these cyanotoxins, the order of concentration and frequency of analytical detection being wing?>?breast?>?head. Consistent with the presence of the microcystins and anatoxin-a in gut contents and the livers of the dead birds and negligible in vitro adsorption to feathers, the cyanotoxins associated with the feathers of the dead wild flamingos are inferred to be primarily of dietary origin.     

Analysis,accumulation and central effects of trihalomethanes. I. bromoform†

The paper presents the results obtained in a study on the effects of organic micropollutants on the central nervous system of the rat, used herein as a suitable model for neurotoxicologic work. Bromoform, a member of the trihalomethane (THM) family, was the pollutant selected since it becomes a characteristic marker in the Barcelona potable water supply.

Analytical data on the GC‐ECD separation of THMs from other volatile halocarbons is presented. A mixed packed column 10% SE‐30/PEG 20M (94:6) at 70°C provides an adequate separation. The distribution of bromoform in rat plasma and tissues showed an accumulation in brain, kidney and fat, increasing in this order and a rapid elimination after 30 min. The effect on the central aminergic metabolism is studied by HPLC. The increased metabolic activity detected could be attributed either to an increased nervous activity, stimulating the metabolism of serotonin, or to its greater degradation.     

Photoreduction of parathion ethyl †

Photodegradation of parathion ethyl in presence of unsaturated biomolecules takes place by sufficient reduction of the phenyl nitro group. Main products are N,0‐dialkenyl hydroxylamino, cis/trans azo, and azoxy derivatives of parathion.     

Mutagenic and carcinogenic effects of cadmium†

There is limited evidence to suggest that certain compounds of cadmium may be mildly mutagenic. Some compounds of cadmium have given rise to fibrosarcoma and rhabdomyosarcoma following subcutaneous or intramuscular injection in the rat and also to interstitial cell tumours of the testis as a sequel to cellular damage and testicular atrophy. Cadmium chloride aerosol inhalation has induced a high incidence of dose‐related lung cancer in the rat. Epidemiological observations suggest that mortality from prostatic and possibly from lung cancer has been increased in cadmium workers who had experienced the very high levels of cadmium, mainly as oxide dust or fume, which existed in the past. The more recent epidemiological studies have not added further evidence in support of the carcinogenic role of cadmium.     

Influence of phytoplankton on complexing capacity of seawater†

Differential Pulse Anodic Stripping Voltammetry (DPASV) was employed to measure Apparent Copper Complexing Capacity (L_t) and Stability Constants (K_s) of related cupric complexes of cell exudates of the phytoplanktonic alga Dunaliella salina. Three natural seawater samples collected 0.1, 2, 5 miles off the Tyrrhenian coast were used as culture medium. The measurements were carried out 1h and 7 days after the inoculum. Furthermore, on one culture the medium was passed through C₁₈‐SEP‐PAK cartridges in order to separate the cell exudates. The results show that Dunaliella salina influences the value of L_t but does not affect the value of K_s . The C₁₈SEP‐PAK retain only 50% of the ligands produced by the cells and capable of complexing copper.     

Enhancement of UV and chromate mutagenesis by nickel ions in the Chinese hamster HGPRT Assay †

1. The HGPRT (Hypoxanthine‐Guanine‐Phospho‐Ribosyl‐Transferase) assay with Chinese Hamster V79 cells was used to measure the mutagenic effects of UV irradiation, potassium dichromate and nickel chloride. The agents were tested separately and in the combinations of UV plus nickel and dichromate plus nickel.

2. UV, Cr(VI) and Ni(II) were confirmed to be mutagenic in the V79 cell assay. The combination of UV(5J/m²) and Ni(II) (0.5 mM) caused a mutation rate 11.2 times above that corresponding to the sum of the individual mutation rates of these agents. The combined action of Cr(VI) (0.1 mM) and Ni(II) (0.5 mM) produced a mutation rate 2.8 fold above that corresponding to the sum of the individual rates of the separate agents.

3. The enhancing effect of nickel chloride on the mutagenicity of UV or Cr(VI) is interpreted by an interference of Ni(II) with the repair of DNA lesions.     

Biogeochemistry of aromatic hydrocarbons in the benthos of microcosms†

Investigations into the fate of petroleum compounds in the marine environment were carried out using experimental microcosms of two sizes and designs. Aromatic hydrocarbons or No. 2 fuel oil were spiked to the water of a 13 m³ continuous flow system and to a 2281 recycled flow system. The transport and alteration of this oil was traced in the sediment and benthic organisms (Glycera americana, Crepidula sp., and Nephtys incisa) of these microcosms. Measurable contamination was found in both sample types. The aromatic hydrocarbon distribution, including relative isomeric distribution (e.g., C₂‐phenanthrenes) was found to be different in sediment and in organisms from that which was originally introduced to the experimental microcosm. Differences in isomer distribution between Glycera and Crepidula were also detected. Based on the experimental data: molecular weight and specific isomeric form, biochemical processes, solubility, and particle adsorption/desorption influence the fate of petroleum compounds in benthic ecosystems.     

The polymorphism of the human serum paraoxonase †

The activity of paraoxonase, the enzyme which hydrolyses paraoxon, 0,0‐diethyl‐0–4‐nitrophenylphosphate, in human serum shows a genetically determined polymorphism with strong interethnic differences. The serum paraoxonase genotype has a significant influence on the paraoxon clearance. Individuals with high serum paraoxonase activity may be better protected against the toxic effects of parathion (0,0‐diethyl‐0–4‐nitrophenylthiophosphate).

In Caucasians the polymorphism is governed by two alleles. The first allele has a gene frequency p _low of 0.67 to 0.78, and is manifested in both the form of a first homozygotic group with low activities and a second heterozygotic group with medium activities. About 50% of all Europeans belong to the low activity group. The second allele with a gene frequency q _high of 0.22 to 0.33 is manifested in the second heterozygotic and a third homozygotic group with medium resp. high activities. The Hardy‐Weinberg rule for a two allele model is valid for the distribution.

The percentage of the low activity group decreases as one moves from Europe to Africa and Asia. In most of the Mongoloids and Negroids only 5 to 20% of the population can be included in the low activity group, which is not even demonstrable in Aborigines, Maoris, Tonga and some African and Indian (Central America) tribes. The validity of the Hardy‐Weinberg rule for a two‐ or three‐allele model must be rejected in non‐Caucasians.     

Bioavailability and metabolism of hexavalent chromium compounds †

Intratracheal instillation of ⁵¹CrCl₃ in anaesthetized rabbits resulted in partial absorption. In blood, the absorbed material was entirely confined to the plasma compartment. Only trace amounts were deposited in liver and kidney. By contrast, after similar application of Na, ⁵¹CrO₄ the bulk of blood radioactivity was present in red blood cells (RBC). Substantial deposition occurred in liver and kidneys. It is concluded that Cr(VI) may enter the body unreduced via the lung and is partially deposited in cells over a prolonged period of time.

Since chromium was accumulated in liver after administration of Cr(VI) we investigated the intracellular disposition of Cr(VI) in the isolated perfused liver. No significant sex differences in chromium distribution were observed. At the end of the experiments (1 h), 60% of the applied dose (312μg Cr/liver) was located in the cytosol, whilst 14% was in the mitochondria, 9% in the microsomal pellet and 2% was associated with the nuclei. Gel chromatography of the cytosolic compartment showed that the overwhelming part of chromium was eluted in fractions with an apparent molecular weight of 6,000 dalton. These fractions exhibited absorption maxima at 410nm and 548nm. It is concluded, that cytosolic reduction might be the main intracellular redox pathway for chromates. This view was confirmed by monitoring the reaction of Cr(VI) with GSH in vitro. GSH reduced Cr(VI) without further cofactors under formation of GSH‐chromium complexes, which possibly represent major intermediates in the metabolism of Cr(VI).     

Alkylation of metals and the activity of metal‐alkyls†

The biochemical basis for resistance to metal ion toxicity is emerging though it is complicated by the different resistance mechanisms. Several strategies for resistance to toxic metal ions have been identified:

1. The development of energy driven efflux pumps which keep toxic element levels low in the interior of the cell. Such mechanisms have been described for Cd(II) and As(V).

2. Oxidation (e.g. AsO^2‐ to AsO₄ ^3‐) or reduction (e.g. Hg²⁺ to Hg⁰) can enzymatically and intracellularly convert a more toxic form of an element to a less toxic form.

3. The biosynthesis of intracellular polymers which serve as traps for the removal of metal ions from solution such as traps have been described for cadmium, calcium, nickel and copper.

4. The binding of metal ions to cell surfaces.

5. The precipitation of insoluble metal complexes (e.g. metal sulfides and metal oxides) at cell surfaces.

6. Biomethylation and transport through cell‐membranes by diffusion controlled processes.

In this short review I shall discuss the implications of biomethylation as a detoxification mechanism for microorganisms as well as for certain higher organisms.     

The interethnic differences of the human serum paraoxonase polymorphism analysed by a quantitative and a qualitative method †

The polymorphism of the human serum paraoxonase¹ was analyzed by two distinguished methods in six different ethnic groups (Caucasians, Mongoloids, Negroids), using (1) the Computer Method² and (2) the Carro‐Ciampi Method³‘⁴. Analysis of the response of the enzyme activities to salts resulting in low and high activity ratios.

Comparison of the results:

In Caucasians we distinguished three phenotypes by the Computer method. The polymorphism was governed by two alleles. The Hardy‐Weinberg rule for a two‐allele model was valid. Individuals belonging to the homozygotic group with low activity had a low activity ratio (Carro‐Ciampi method). With both methods a frequency between 57% and 61% was observed for this group. Individuals with medium and high activity had a high activity ratio.

In Negroids and Mongoloids samples we found (by the Computer method) a low activity group (Ghanaians 9.6%, Jamaicans 13.6%, Indonesians 6.7%, Koreans 19.6%). The Hardy‐Weinberg rule for a two‐ or three‐allele model was not valid. Individuals belonging to the low activity group had a low activity ratio, all individuals with higher activity a high activity ratio (Carro‐Ciampi method).

Our results suggest that the members of the low activity group in the three races are homozygote for an identical allele.     

Multielement techniques for determination of Inorganics in Watert†

Several analytical techniques offer the capability of simultaneously determining 5–40 elements. Of those techniques available, two are receiving extensive attention for analysis of water samples. Optical emission spectrometry utilizing plasma excitation has now evolved to a status in which 20–40 elements can be simultaneously determined on a routine basis. Instrumentation which relies on ion exchange chromatography separations is available which allows the routine determination of several cationic and/or anionic species.

The present discussion will briefly describe each of the above multielement analysis systems, summarize their respective capabilities and limitations with respect to water analysis, and project areas of future development.     

Source and Distribution of Heavy Metals in Food†

Diet is the major route of exposure to contaminants, therefore food monitoring programs are essential to estimate the degree of human risk. We present a study undertaken in Austria dealing with the total uptake of mercury, cadmium, arsenic and lead. The dates were calculated from diet as well as from commercial food. Higher contaminated foodstuffs are registered and a comparison was made between body burden of babies and adults on the basis of bodyweight. It was pointed out, that cooked or washed food ready to eat has a lower burden of lead and cadmium than non prepared food due to household processing, which may decontaminate food to some extent’ Arsenic content was not reduced, because it is bound chemically and not primarily deposited on the surface of food such as in the case of lead. It was mentioned that the type of compound plays an important role in the assessment of action of those elements on humans.

Their biological activity is not necessarily related to their concentration in food. As in the case of selenium and chromium one has to take into consideration that there is a narrow range between benefit and harm, toxicity, and minor supply.

Therefore we conclude, that due to interactions of elements, their different biological activity and due to the difference of compounds present in food, we have to relativate the terms toxicity and essentiality. Then we can get a more profound insight into the problems of trace elements in food.     

Biological effects of selenium and relationships with carcinogenesis†

Selenium is an element that is both essential, at low levels of dietary intake, and toxic, at high levels, to man and a wide range of animals. The purpose of the communication is to review the evidence for the involvement of hyper‐ and hypo‐selenosis in the aetiology of cancer. High levels of selenium intake have been blamed for an increase in cancer, dental caries and an array of other conditions. Critical evaluation of the evidence, however, leads to the conclusion that such claims have little substance. On the other hand, low levels of selenium intake may be associated with an increased incidence of certain forms of cancer and there appears to be an inverse relationship between blood selenium levels and cancer death rate in the United States. Keshan Disease, in the People's Republic of China has only been shown to occur in populations severely depleted of selenium. In well nourished populations there is no evidence to indicate that selenium supplements have any prophylactic or therapeutic benefit against human diseases such as cancer, cardiovascular disease or cystic fibrosis.     

Selective cellular and molecular effects of beryllium on lymphocytes†

An involvement of the immune system has been implicated in many of the whole animal effects of beryllium (Be) and its compounds and thus lymphocytes are a relevant cell model to study some of the cellular and molecular actions of the metal. Recent results indicate micromolar concentrations of Be salts are weakly mitogenic to lymphocytes in vitro probably as a result of direct interactions with the cell membranes. Furthermore evidence has been obtained that high affinity lymphocyte Be‐binding may be predictive of the potential to express delayed Be hypersensitivity in vivo. Although sub mitogenic concentrations of Be salts may actually activate accessory (macrophage type) cells, concentrations normally mitogenic to lymphocytes were found to inhibit their proliferative response to accessory cell dependent mitogens as a result of a Be²⁺ concentration dependent destruction of the adherent cell population. Higher concentrations of Be salts proved cytostatic to lymphocytes possibly by the established property of intracellularly accumulated Be²⁺ to inhibit cell division. Lymphocyte proliferation can be related to increases in protein phosphoryl‐ation capacity which in vitro can be inhibited by Be²⁺ and preliminary evidence indicates that tyrosine phosphorylation may be particularly sensitive.     

Environmental impact assessment for aquatic ecosystems of industrial effluents†

A system of environmental impact assessment of industrial effluents is presented and discussed. Environmental assessment consists of three phases, hazard assessment, risk determination, and societary evaluation. It is regarded an integrated study of natural sciences, i.e. ecochemicology (chemical fate), ecotoxicology (toxical effects), and ecoepidemiology (biological damages); and social sciences, e.g. geography, sociology, and economy. And the outcome should be the basis of political decision making. Hazard assessment may be divided into a subphase of hazard identification from a base set of scientific information on the industrial effluent, and a second subphase of hazard analysis at a more elevated level of scientific information akin to the procedures suggested elsewhere for single chemicals. Methods of hazard assessment are laboratory experiments, e.g. chemical analysis, degradation and bioaccumulation studies, acute and chronic toxicity assessments, physical modelling, etc. Risk determination may then be conducted by extrapolation of the obtained results from laboratory to field, i.e. by risk estimation to the receiving waters, using methods of hydraulic modelling, biological surveillance, etc. Risk‐benefit evaluation is conducted in the society evaluation phase by balancing environmental consequences against the society value of the production. The outcome is determined by the interaction between hazard‐makers (industry), risk‐takers (administrators), guardians (regulators), and assessors (scientists) in risk management, the crank of environmental assessment.     

Investigation of the sources of aliphatic hydrocarbons in the mussel Mytilus edulis from north sea oil production platforms by Capillary glc and CGCMS†

Aliphatic hydrocarbons isolated from mussels collected over a 20 month period from three North Sea Forties field oil production platforms have been examined by capillary gas chromatography and gas chromatography‐mass spectrometry.

The biological hydrocarbons consist predominantly (300–700 μh g^‐1 lipid) of C₃₁ and C₃₃ n‐alkenes with 2, 3 and 4 double bonds probably derived from a dietary intake of Emiliania huxleyi, a microscopic coccolithophorid alga. In some of the mussels C₁₈, C₂₀ and C₂₂ n‐alkanes are present in unusually high abundance compared to their odd carbon number homologues. Possible reasons for this distribution are discussed.

The concentrations of fossil fuel hydrocarbons (mainly unresolved complex mixtures) in the mussels ranged from 330 to 5,298 μg g^‐1 of lipid. No significant increase in values was detected two months after the start of discharge of treated Forties production water from the Forties D platform.

To determine the pollutant hydrocarbon sources, the sterane and terpane distributions of hydrocarbon fractions isolated from the mussels and from a number of fossil fuels, including Forties crude oil, were examined by mass fragmentography. The steranes in the mussels from the Forties C and D platforms contained higher proportions of regular 14α(H), 17α(H) components than Forties oil in which the steranes were mainly 13ß(H), 17α(H) diasteranes. In addition, the terpane distributions of the mussel fractions differed markedly from that of Forties crude oil, particularly in the relative abundance of diterpanes to triterpanes, which was higher in the mussel fractions than in the oil, and in the absence of 17α(H), 18α(H), 21ß(H)‐18, 30‐bisnorhopane, a known component of Forties crude. Furthermore, the ratio of C₂₉ to C₃₀ triterpanes was greater than unity in the mussels but much less than unity in the Forties oil. These data preclude Forties or other similar North Sea crudes as the major source of pollutant aliphatic hydrocarbons in the mussels. The relative concentrations and distributions of steranes and triterpanes suggest that the most likely source of pollutants is a Middle Eastern based oil derived either from rig activities, or from background pollution in the North Sea. The low concentration of ≥ C₂₇ steranes in the more polluted mussels (e.g. Forties B; UCM > 200 ppm dry wt.) suggests that gas oils used on the platforms may be the major source of petrogenic hydrocarbons in these samples.     

Genetic toxicology of metal compounds: An examination of appropriate cellular models †

Bacterial systems have not had success predicting metal carcinogenicity. Hypotheses explaining this failure are examined. Using a broad genetic endpoint, λ prophage induction, under sub‐toxic growing conditions, genotoxicity is seen for compounds of chromium, manganese, lead, molybdenum and tungsten. Copper, manganese, arsenic and molybdenum compounds enhanced UV mutagenesis in E. coli WP2.

The toxicity of metal compounds to cultured mammalian cells correlates well with rat oral LD₅₀ values. Whereas insolubility can present problems in bacterial studies, concentrations of metal compounds toxic to mammalian cells can be determined even in the presence of precipitate, and sometimes [Pb(NO₃)₂, BaCl₂ and BeCl₂] occurs only in its presence. PbS and MnS, which are insoluble, are much more toxic than the more soluble compounds Pb(NO₃)₂ and MnCl₂. These results demonstrate the importance of cellular phagocytosis of insoluble metal compounds as a factor in studying the toxicity and genotoxicity of metal compounds.     

Biochemical properties of beryllium potentially relevant to its carcinogenicity†

The carcinogenicity of beryllium to several animal species is well established and evidence exists which strongly suggests that this is the case in human exposure. In this review several biochemical properties of the metallocarcinogen are considered including, the causation of cell transformation, and infidelity of DNA synthesis, inhibition of cell division and enzyme induction, and interference with regulatory mechanisms controlling gene expression. These effects are discussed in relation to beryllium chemistry, cellular accumulation mechanisms and distribution to subcellular organdies and molecular targets. It is suggested that the ultimate location and interactions of the metal ion in cell nuclei and its selective inhibition of certain protein phosphorylation reactions in particular are the biochemical effects potentially most relevant to induction of beryllium carcinogenesis.