摘要:

Towards sustainable pesticides Focus J. Environ. Monit., 1999, 1 33N In the light of increasing concern over the environmental impact of pesticides, we look at the evolution of national and international policies for sustainable pesticide use, and the associated data requirements. Few issues are as divisive as pesticides. Since the publication of Silent Spring, Rachel Carson’s landmark work on chemicals in the environment, published in 1962, pesticides have become something of a touchstone in the environmental debate.For environmentalists, pesticides have come to symbolise our over-reliance on chemicals of which we know little and care less. For industrialists, they epitomise the increasing scrutiny being extended to chemical products in all areas of modern life.Pesticides remain of central importance today, touching, as they do, many central issues in the sustainability agenda. In waste management, water quality, chemical hazards, product labelling, sustainable agriculture and third world development, pesticides retain a prominent role. What, then, are the prospects for a more sustainable approach to pesticide use, and how can environmental monitoring inform such policies? Pervasive pollutants Pesticides—the term is used here to include categories such as insecticides, herbicides and other agricultural chemicals—are pervasive in modern society.They are important to modern agriculture to control pests, weeds and diseases. They also have significant nonagricultural uses, such as the treatment of textiles, timber preservation, and the fouling protection of boats.Around 500 pesticides are in common use, many of them either known or thought to be hazardous.1 The main environmental impacts arise from the high potential to pollute water courses, through direct run-oV, spray drift or shortcomings in storage, handling or disposal. They vary greatly in their chemical and physical characteristics, and thus in their solubility in water.However, pesticides are, by their nature, highly active in that they have a high potential to kill specific organisms. Concerns over health eVects focus around three issues: (1) Persistence: many of the chemicals are known to be highly resilient and to accumulate within the environment. Even where they do degrade, their breakdown products may also be toxic to some animals and plants.(2) Low doses: for some chemicals, there is increasing evidence of health impacts through long-term exposures at very low dose levels. These long-term eVects are thought to include cancer, reproductive changes, neurotoxicity and endocrine disruption. (3) Multiple interactions: while attention to date has focused mainly on single species, there is increasing evidence of detrimental eVects from lowlevel exposures to mixtures of diVerent chemicals—known as multiple chemical sensitivity (MCS).Dangerous cocktail Taken together these factors amount to a significant threat. So much so that Professor Nicholas Ashford of Massachusetts Institute of Technology, a leading environmental scientist and health adviser to the United Nations, warned recently that ‘pesticides are the most serious problem we have today in the industrial countries’.2 Professor Ashford is particularly concerned about MCS.The huge rise in pesticide use over the last fifty years could be responsible for a variety of illnesses with common symptoms, such as skin rashes, breathing problems, cancers and birth defects. According to Professor Ashford, sensitised patients have been found to react to levels of chemicals so low that they are undetectable with existing testing methods.Unable to diagnose a classic cause-and-eVect, the resulting symptoms baZe doctors, who frequently attribute the condition as a mental illness. In his latest book, he advocates that this sensitisation can be explained by a new theory called toxicant induced loss of tolerance (TILT).3 The MCS theory is still contentious, but according to Ashford there is now compelling evidence that this condition should be recognised as a separate illness.‘The last seven years of research has not furthered the case for psychosomatic origins at all’, he says, ‘but it has definitely furthered the case for physiological origins’.2 The shortcomings in current toxicological testing procedures have been underlined by recent research from the University of Wisconsin (see Box 1).4 A five year study suggests that combinations of commonly used agricultural chemicals, in concentrations similar to those found in groundwater, can significantly influence immune and endocrine systems, as well as neurological health.Study leader Warren Porter said the single most important finding was that ‘common mixtures, not the standard onechemical- at-a-time experiments, can show biological eVects at current concentrations in groundwater’.The study focused on three farm chemicals in common use: aldicarb, an insecticide, atrazine, a herbicide, and nitrate, a chemical fertiliser. Risk reduction strategies As well as health and environmental aspects, the social regulation of pesticide use is driven by other issues.5 These include: a more general concern about agriculture’s dependence on and heavy use of chemical inputs; and a new social mandate to incorporate environmental goals into agricultural policy and practice. The development of pesticide/ herbicide resistance by some pests and crops, and the increased use of pesticides that has accompanied the intensification of agriculture, have been of particular concern.Over recent years policy-makers have sought coherent frameworks for this highly complex agenda—so-called pesticide risk reduction strategies. These generally aim to balance the triple objectives of: promoting sustainable agriculture, achieving high levels of environmental and health protection, and maintaining free trade.A wide variety of instruments are employed. A 1996 survey found that34N J. Environ. Monit., 1999, 1 pesticide registration provided the foundation for pesticide risk reduction in all OECD countries.5 Registration is the process by which pesticide producers submit data to governments on the potential health and environmental risks of specific products, and governments evaluate the data to determine whether or not to allow the products on the market.Such processes tend to be extremely slow and resource intensive, and have not been helped by the need for old pesticides to be re-registered. Other measures commonly applied are: $ Reducing consumer risks, through setting and monitoring of residue levels; $ Reducing risks to workers, through occupational exposure levels, certification programmes, education and training, information campaigns, safety regulations etc.$ Reducing risk to the general environment, through emission limits, environmental quality standards, and special protection for sensitive species and habitats. Additional measures are aimed at reducing pesticide use or patterns of use, often as part of national reduction programmes.These include: $ Increasing eYciency and eVectiveness, through activities to help farmers adopt best pest control practices. $ Implementing integrated pest management (IPM), based on the use of biological pest control agents and/or pest-resistant varieties. IPM usually requires intensive guidance, information and demonstration actions. $ Economic instruments as a stimulus to reduce unnecessary use.Such instruments include subsidies for using IPM and organic methods; promotion of green labelling as a means of market diVerentiation; and introduction of taxes on pesticide sales. Policy trends Recognising that registration is a bottleneck, but also a key policy tool, an extensive international co-operation is being established, under the auspices of the OECD.Last year OECD countries agreed a set of common guidelines aimed at bringing their registration procedures closer together.6 The guidelines establish two new formats— one for industry in making data submissions, the other for governments when writing their evaluations. The new formats do not require countries to make the same regulatory decisions. Rather they aim to minimise duplication of eVort by industry and governments in pesticide registration and to improve the quality of evaluation reports. Further work is underway to improve the sharing of information between countries on pesticide risk assessments.There is also increasing recognition that eVective pesticide risk reduction policies need to go beyond safety assessments for individual chemicals prior to market authorisation.The European Commission last year presented the results of a six-year study into future options for a sustainable pesticides policy.7 Among the project’s findings, contained in a 3000 page consultancy report, were that, for a given crop, pesticide use varies widely between regions, for no apparent reason, and residues in drinking water sources sometimes exceed the EU limit of 0.1 mg l-1.The project found no or very limited economic benefit in using pesticides beyond the level for integrated production, estimated to be 25–40% lower than current levels for the standard farm. The report recommended potential EU involvement in monitoring the application and environmental impact of pesticides, inspecting spraying equipment and supporting farmer training.There is also broad agreement that the EU’s programme for risk assessment, under the 1991 pesticide authorisation directive, is moving too slowly. A refocusing of resources is called for around a priority list of the most problematic chemicals. Sustainable pesticide use should also be integrated into agriculture and water policies.New initiatives, based on these proposals, will be set out in a policy paper from the Commission to be issued later in the year. Europe weighs up taxes As yet it is not clear whether the Commission will give its backing to EU-wide pesticide taxes. While the Commission has no powers to set taxation levels—a power reserved for the Member States—it could propose EU-wide frameworks on environmental Focus Box 1: Shortcomings in pesticide testing According to some commentators, current testing methods for the registration and use of pesticides are fundamentally flawed, in that they fail to address conditions under which mixtures of chemicals interact.Important deficiencies in current testing protocols include: $ Unrepresentative formulations: Tests tend to use pure forms of pesticidal active ingredients rather than commercial formulations.Most tests are missing three types of chemical additives: contaminants of manufacturing processes, toxic waste from reactor cleaning processes, and ‘inert’ ingredients. $ Single exposure routes: Standard toxicological tests only evaluate one route of exposure at a time, rather than all possible routes (oral, cutaneous and respiratory).$ Non-carcinogenic eVects: Toxicological tests have typically focused on cancer and mutation end-points, at the expense of other critical concerns, such as endocrine and immune systems. $ Pulse dose eVects: Current tests fail to evaluate low dose pulse exposure. Pulse doses of low levels of pesticides may damage a foetus at critical times in the development cycle.This is because the embryo has almost no defensive systems against chemicals and no feedback systems to modulate chemical concentrations in early development. $ Multiple interactions: Current testing requirements fail to evaluate exposure eVects from chemical mixtures. While it is impossible to examine all possible mixtures, common combinations likely to arise from crop rotation and tillage practices could be examined. $ Environmental conditions: Laboratory animals generally live in a carefully controlled environment.Researchers know that diVerent responses can occur under stress. Adapted from ‘Endocrine, immune and behavioural eVects of aldicarb (carbamate), atrazine (triazine) and nitrate (fertilizer) mixtures at groundwater concentrations’, Warren Porter et al., Toxicology and Industrial Health, 1999, vol. 15, pp. 133–150.J. Environ. Monit., 1999, 1 35N in September, and UNEP is optimistic of finalising the agreement by next year. Moreover, this is only one of many international fora involved in pesticide regulation. Mechanisms such as ‘IRPTC’, ‘PRTR’, ‘GEENET’, ‘IPCS’ all occupy particular roles: the whole field is a veritable alphabet soup.11 Despite this intensive international activity, what is striking is not how much we know about these chemicals, but how little.Take, for example, lindane, a persistent organochlorine insecticide in use since the 1940s. A recent (leaked) report from the European Commission, prepared by the Austrian government, points to potentially serious health eVects.12 Based on a review of the scientific literature, the report notes associations with cancer and birth defects, and toxic eVects on blood and genes.However, the report’s main conclusion is that despite fifty years of commercial use, there is a lack of basic scientific information about the impact of lindane on health and the environment.Multiply this by the many hundreds of pesticides in common use, and the scale of our knowledge gap becomes truly alarming. Informing policies Against this background of pervasive use and a rapidly evolving policy agenda, there is an urgent need for reliable information. The analytical sciences have to provide a more comprehensive picture of the risks posed by pesticides—and find ways of communicating this beyond the scientific community.Information programmes need to address three purposes: (1) Product risk assessment: policymakers need comprehensive reliable information to inform product registration and re-registration. The systematic assessment procedures and data-sharing exercises being promoted by OECD should go some way towards achieving this.(2) Post-approval monitoring and surveillance: in most developed countries relatively little eVort is devoted to the monitoring of pesticides’ environmental impacts once in use, and where programmes do exist there is often a lack of transparency between the methodologies used. There is a need for more strategic approaches, similar to those being pursued for product registration.The EU is to explore new sources of funding for post-approval monitoring. (3) User-friendly indicators: Securing the commitment of users and other stakeholders to pesticide reduction requires clear and meaningful indicators by which to measure progress. Given the complexity of the issues, this is a demanding requirement—but an essential one. Scientists should engage in this process and bring their own perspectives to it.The UK, for example, has already proposed potential indicators, while recognising that they are not always scientifically rigorous.13 They include concentrations in rivers and groundwater, quantity of active ingredients used, total spray area, and number of wildlife incidents. These data processes should, in turn, reflect and inform key policy themes.Four, in particular, stand out. Firstly, there is the MCS debate. In the light of a growing body of evidence on the inadequacy of current testing protocols, new methods are required which take account of low dose exposures (including short, so-called ‘pulse doses‘) and chemical mixtures. Secondly, the implications of genetically modified crops for pesticide use is unclear.The agrochemicals industry claims that genetically modified herbicide tolerant (GMHT) crops will reduce herbicide use. However, there are indications that use of some herbicides, such as glyphosate, is increasing following the introduction of genetically engineered glyphosateresistant crops (see Box 2).14 With independently verified data hard to come by, there is a need for more rigorous scientific investigation of environmental impacts.The level of pesticide residues in food is another major health issue.15 A recent survey looking at seven pesticides in fruit and vegetables found much higher levels than expected (see JEM, 1999, 1(1), 10N). In the US, maximum residue levels (MRLs) are regulated at federal level by the Environmental Protection Agency and the Food and Drug Administration, with information on the regulations and limits readily available on the EPA and FDA websites.This contrasts sharply with the EU where MRLs are still set by the Member States—in some cases under a great deal of secrecy. The levels need to be harmonised and the process made more transparent. Finally, better information should help reinforce current trends towards cross-party stakeholder initiatives.In the UK, for example, indicators of pesticide charges to safeguard the Single Market. Charges are already high on the agenda in several European countries. Belgium, Denmark and Sweden already have pesticide charging systems, while the Netherlands and the UK are considering them. Norway is set to introduce an environmental levy on pesticides later this year, under a fouryear action plan.8 The scheme involves a fixed levy of 13 Nkr (1.74US$) per hectare plus a diVerentiated levy based on health and environmental hazard.Products sold to farmers and other professional users will be divided into five hazard/risk classes, based on the weight of active ingredient. In March of this year, the UK government invited consultation on similar proposals for hazard-based tax rates.9 To date, the most rigorous regime against pesticides is that adopted in Denmark.The government has long advocated reduced spraying frequencies for agricultural use, and recently proposed a ban on the use of pesticides in private gardens (see JEM, 1999, 1(2), 26N). Further measures were announced in March, following the publication of an advisory study into how to reduce environmental and health risks.10 The Bichel report, named after its author, recommends a three-pronged strategy of cutting the application frequency; creating low or no-spray zones near drinking water catchment areas; and encouraging organic farming.In Denmark, as elsewhere, national targets used to be based solely on weight of pesticides used in each spray.Spray frequency is now seen as a more appropriate indicator, since it correlates better with environmental impacts as pesticide active ingredients become more concentrated. Mind the gap While even Denmark has stopped short of an outright ban, the writing is already on the wall for many common pesticides under legislation aimed at managing persistent organic pollutants (POPs).Under the auspices of the United Nations Environment Programme (UNEP), representatives of over 100 countries are negotiating an international treaty to reduce and/or eliminate releases of certain POPs (see JEM, 1999, 1(1), 10N). Of the 12 specific POPs proposed under the first stage, eight are pesticides and many more could be added under the treaty’s pollutant identification procedure.Participants will meet again in Geneva Focus36N J. Environ. Monit., 1999, 1 London. Available at: www.gn.apc.org/ pesticidestrust 3 N. Ashford and C. S. Miller, Chemical Exposures: low levels and high stakes, 2nd edn. Wiley, New York, 1998. 4 W. Porter et al., Endocrine, immune and behavioural eVects of aldicarb (carbamate), atrazine (triazine) and nitrate (fertilizer) mixtures at groundwater concentrations, Toxicology and Industrial Health 1999, 15, p. 133. 5 Activities to Reduce Pesticide Risks in OECD and Selected FAO Countries I, Part 1: Summary Report, OECD Environmental Health and Safety Publications, Series of Pesticides No. 4, OECD, Paris, 1996. Available at: www.oecd.org/ehs 6 The guidelines can be accessed at: www.oecd.org/ehs 7 Proceedings of the Second Workshop on a Framework for the Sustainable Use of Plant Protection Products in the European Union, European Commission, 1998.Available at URL: europa.eu.int/ comm/dg11/ppps/home.htm 8 Norway launches ambitious risk reduction plan in Pesticide Risk Reduction News, (1), January 1999, OECD, Paris. Available at URL: www.oecd.org/ehs 9 Design of T ax/Charge Scheme for Pesticides: Consultation Document, Department of the Environment, Transport and the Regions, 1999.Available at: www.env.detr.gov.uk 10 Denmark to further cut farm pesticides use, in ENDS Daily 23rd March 1999. See www.ends.co.uk. 11 PRTR is the Pollutant Release and Transfer Register; GEENET is the Global Environment Epidemiology Network; IPCS is the International Programme on Chemical Safety. 12 T he end for lindane, in Pesticides News (43),March 1999. Pesticides Trust, London. Available at: www.gn.apc.org/ pesticidestrust/pn43.htm 13 Development of a set of indicators for sustainable agriculture in the United Kingdom: A consultation document, Ministry of Agriculture, Fisheries and Food, London.June 1998. Available at: www.maV.gov.uk 14 Scientific Review of the Impact of Herbicide Use on Genetically Modified Crops, December 1998, Pesticides Safety Directorate, Ministry of Agriculture, Fisheries and Food, London. See also, Resistance to glyphosate in Pesticides News (41), September 1998, Pesticides Trust, London. Available at: www.gn.apc.org/pesticidestrust 15 For a brief review of international experience see Pesticide residues variability and acute dietary risk assessment, Pesticide Safety Directorate, Ministry of Agriculture, Fisheries and Food, London.Available at: www.maV.gov.uk/aboutmaf/agency/psd/ psdhome.htm 16 The UK Pesticides Forum. See: www.environment.detr.gov.uk/ pesticidesforum/ 17 For current status of analysis techniques see Proceedings of the First International Conference on Predicting the Environmental Fate of Agrochemicals, 26th–27th May, London, UK.Available from IBC Global Conferences, www.ibcuk. com/IV140 Mike Sharpe use for ‘sustainable agriculture’ are being actively supported by stakeholder groups which bring together manufacturers, farmers, NGOs, regulators and others.16 New approaches and techniques As they contemplate these requirements, analysts have a variety of new approaches and techniques at their disposal.These include geographical information systems (GIS), fuzzy logic, artificial intelligence and statistical modelling (based on Bayesian and other methods).17 Over the last five years, GIS, in particular, has made significant contributions to the analysis of pesticide exposure.GIS techniques are now being extended to cover pesticide fate in the environment. Coupling of GIS with modelling enables the use of geographically based input data as well as spatial mapping and analysis of simulation results. Notes 1 For general information on environmental and health eVects of pesticides see sources such as: EPA OYce of Pesticide Programs, www.epa.gov/pesticides/info.htm; Rachel Carson Council, members.aol.com/ rccouncil/; and Pesticide Action Network, www.panna.org 2 Quoted in Pesticides News, No. 41, September 1998. Pesticides Trust, Focus Box 2: Herbicide-tolerant crops in North America In 1996, 2.8m ha of GM crops were grown in North America, of which 0.64m ha (23%) were herbicide-tolerant. By 1997 this increased to 0.7m ha, or 54% of all GM crops. In 1996, for example, only 2% of the North American soybean crop was glyphosate-tolerant. This increased to nearly 15% in 1997, 30% in 1998, and an estimated 60–70% in 1999. Although both glyphosate and glufosinate are active against a wide range of weeds, there is evidence of varying eVectiveness. In Iowa, weeds that emerge later were found to appear more often because they avoid the planned glyphosate applications. Moreover, glyphosate is not eVective against all weeds at the application rates for which farmers are willing to pay. In practice, other herbicides are sometimes tank-mixed with glyphosate or are used in sequence. This is primarily to control the weeds that glyphosate does not control well, rather than to delay the onset of resistance. As prices fall, farmers may choose to use more glyphosate, either through a higher application rate or multiple applications. Adapted from Scientific Review of the Impact of Herbicide Use on Genetically Modified Crops, Pesticides Safety Directorate, Ministry of Agriculture, Fisheries and Food, UK. December 1998.

ISSN:0960-7919    

DOI:10.1039/a903465j

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

J. Environ. Monit., 1999, 1 37N US Focus Perchlorate rockets to US national attention Ground and surface water contaminated with ammonium perchlorate, a component of solid rocket fuel, has placed drinking water supplies at risk in a number of communities throughout the US, especially in the West. An expedited research agenda is providing many answers about this new environmental problem, but important questions remain.In late 1996, groundwater migration of perchlorate was discovered in California following an improvement in the ion chromatographic (IC) method used for its detection.1 The discovery caused immediate and intense concern among water utility companies, regulators and environmental groups for many reasons. Although it is a component of rocket fuel, ammonium perchlorate reacts explosively only at elevated temperatures (380 °C); it is stable at 110 °C.2 At high doses, the perchlorate anion is known to block iodide uptake by the thyroid, but the eVects of chronic low-level exposures are poorly defined.This oxidising agent is exceedingly mobile in aqueous systems and can persist for many decades under The image was obtained from IMSI’s MasterClipsA and MasterPhotosTM Premium Image Collection.typical ground- and surface water conditions. In addition, there was, and still is, no proven treatment method for cleaning up contaminated drinking water. To top matters oV, no-one was aware of the extent of the contamination due to unregulated eZuent discharges. Faced with these uncertainties, the California Department of Health Services (CDHS), together with the California Environmental Protection Agency, compiled available health data and set a provisional action level of 18 mg L-1 in drinking water.3 Over 20 water utilities closed wells contaminated above this level.This action prompted local concern about the safety of drinking water. For example, hundreds of people who live near Los Angeles are involved in law suits over health risks believed to be associated with intake of perchlorate and volatile organic compounds.In Nevada, the perchlorate anion has been detected in the Colorado River—waters that are used for drinking, irrigation and recreation by millions of people in the South-West. As a result, the US Congress committed $2 million for research into water treatment methods.The US Department of Defense, a major user of perchlorate, and a coalition of perchlorate manufacturers and users, put up another $2.2 million to fund an expedited toxicological assessment under the oversight of the US Environmental Protection Agency (EPA). Then the EPA began assessing the likelihood of perchlorate contamination throughout the country. These initiatives have begun to pay oV.The EPA produced a draft toxicological risk assessment in December 1998—in just one year rather than the customary seven. There is a now a large body of analytical and chemical data, and interlaboratory validation of the California DHS IC method for water samples is underway.4 Pilot testing for some water clean-up technologies has already started.The extent of the groundwater contamination problem is also better understood. Concentrations ranging from 8 mg L-1 to 3 700 000 mg L-1 have been found. The latter value corresponded to samples collected at a former munitions site. The Colorado River and several California wells show concentrations in the range of 8–30 mg L-1. To date, most of the sites where perchlorate has been detected in ground or surface waters are primarily in areas associated with the development, testing or manufacture of aerospace materials. For the country as a whole, EPA now estimates that perchlorate has been either manufactured or used in 44 states.There are 14 states with confirmed releases in ground or surface waters.5 Huge monitoring task ahead There is still a huge monitoring task ahead because only a small number of water supplies have been monitored using techniques capable of measuring perchlorate at low levels.5 In addition to the interlaboratory validation which is being conducted by the US Air Force Research Laboratory and EPA’s National Exposure Research Laboratory,4 other methods are being developed involving gravimetry, capillary electrophoresis or spectrophotometry.Potentiometric measurements employing ion selective38N J. Environ. Monit., 1999, 1 electrodes (ISE) constitute another approach.6 This method, well established for research, has potential for field monitoring and for continuous monitoring within treatment facilities. Health concerns persist EPA’s new draft toxicological assessment substantiates concerns about the health eVects of chronic exposure to low levels of perchlorate. The December 1998 draft supports an action level of 32 mg L-1, almost double the 18 mg L-1 value that California used to shut down drinking water sources.5 However, an order-of-magnitude uncertainty associated with this standard indicates that the updated draft does not dramatically alter the perchlorate problem.The draft toxicological assessment was based on animal testing expressly designed and conducted to evaluate human health. A final toxicological assessment is due in 2000, on completion of further toxicological assessments (including human exposure) and peer review. Treatment challenges Perchlorate’s chemistry and physical properties prevent the use of standard water-treatment methods.7 Although the perchlorate ion (ClO4-) is the most oxidised form of chlorine that can exist in water, it is remarkably unreactive and its salts are extremely soluble.Common reducing agents do not react with it and common cations do not precipitate it. As a result, standard water treatment methods neither remove nor destroy it. Methods under consideration for the treatment of perchlorate-contaminated waters include: ion exchange, membrane filtration, electrodialysis and biological techniques.Bioremediation and biological or biochemical methods appear to be the most economically feasible, fastest and easiest means of dealing with perchlorate at all concentrations.7 Several genera of microorganisms are capable of using perchlorate as an oxidant for metabolism.8 A significant barrier to the implementation of any biological method is that such techniques have never been used to treat drinking water in the US.A pilot project that uses a biological treatment system to remove perchlorate from groundwater began early this year in Baldwin Park, near Los Angeles, CA.4 No federal regulations There are currently no federal regulations for perchlorate.Perchlorate was placed on the DrinkingWater Contaminant Candidate List in March 1998.5 The list is a source for priority contaminants, defined as either known or anticipated to occur in drinking water, for research, regulatory determinations and monitoring by the states. Perchlorate was listed as a contaminant that required additional research and occurrence information before regulatory determinations could be considered.Action under the Safe DrinkingWater Act would not even begin before 2001,5 but EPA could issue a health advisory if the toxicity results warranted it. Perchlorate is the topic of an upcoming symposium at the American Chemical Society national meeting in US Focus August, 1999, in New Orleans, LA. For details see: http://www.epa.gov/ OGWDW/ccl/perchlo.html Notes 1 California Department of Health Services, Perchlorate in California Drinking Water, September, 1997.http://www.dhs.cahwnet.gov/prevsrv/ ddwem/perch.htm#advice. 2 A. A. Schilt, Perchloric Acid and Perchlorates, The G. Frederick Smith Chemical Company, Columbus, OH, 1979. 3 California Department of Health Services, Determination of Perchlorate by Ion Chromatography, Rev. 0, June 3, 1997. Sanitation and Radiation Laboratories Branch. 4 D. Tsui and S. Pia, Analytical T echniques and Pending Interlaboratory Study, presented at the Perchlorate Stakeholders Forum, May 19–21, 1998, Henderson, Nevada. 5 U.S. EPA OYce ofWater perchlorate web site, Overview of Perchlorate Issues, United States Environmental Protection Agency OYce of GroundWater and Drinking Water, 1998. http://www.epa.gov/OGWDW/ccl/ perchlo.html 6 United States Environmental Protection Agency, Perchlorate Environmental Contamination: T oxicological Review and Risk Characterization. Based on Emerging Information (external review draft), National Center for Environmental Assessment NCEA-1–0503, 1998. http://www.epa.gov/OGWDW/ccl/ perchlor.html 7 E. T. Urbansky, Perchlorate Chemistry: Implications for Analysis and Remediation, Biorem. J., 1998, 2, 81. 8 B. E. Logan, A review of chlorate- and perchlorate-respiring microorganisms Biorem. J., 1998, 2, 69. Rebecca Renner Science writer and editor based in the US Tel:+1 570 321 8640; Fax: +1 570 321 9028, e-mail: applepie@sunlink.net

ISSN:0960-7919    

DOI:10.1039/a903470f

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

J. Environ. Monit. 1999 1 39N News Legislation UK strengthens safety legislation A series of measures which amount to a major overhaul of health and safety legislation have recently come into eVect within the UK. The most significant measure is a more integrated approach to health and safety contained in the Control of Major Accident Hazard (COMAH) Regulations 1999. The regulations implement the requirements of the EU�s �Seveso II Directive� on control of major accident hazards involving dangerous substances. Speaking at an international conference in London shortly before the new regulations came into force Environment Minister Alan Meale said �COMAH takes account of lessons learned from chemical accidents in the UK and abroad. It gives eVect to a tighter more focused preventive safety regime through more detailed safety reports and major accident prevention policies�.COMAH also brings an increased emphasis on controlling risks to both people and the environment through sound management. Guidance for the new regulations will be published in June. The main UK regulation covering occupational exposure limits has also been revised. The Control of Substances Hazardous to Health (COSHH) Regulations 1999 came into force at the end of March and replace the 1994 Regulations. The Schedule listing substances assigned Maximum Exposure Limits (MELs) has been removed from the new Regulation and in future will appear in a separate HSE publication EH40 Occupational Exposure L imits. HSE said the move will enable them to approve and implement changes to the list more quickly. COSHH 1999 also includes various changes in definitions and new MELs for aniline glutaraldehyde benzene and rosin-based solder flux.Some further minor changes to classification and labelling have been introduced by the Chemicals (Hazard Information and Packaging for Supply) (Amendment) regulations 1999. HSE Infoline +44 (0)541 545500 California forced to act on methyl bromide The State of California has been ordered by a supreme court judge to adopt regulations to protect the public against the fumigant methyl bromide. Environmentalists see the decision as a turning point in their long battle against the chemical a highly volatile and acutely toxic pesticide (it is a neurotoxin and causes pulmonary injury). They say that the gas routinely drifts from farm fields into adjacent neighbourhoods and claim that 19 people have died in California from exposure to methyl bromide in structural fumigation.Over 17 million pounds of the chemical is used in California each year more than any other state mostly in the agricultural areas in the centre and south. The order requires the state authorities to adopt new regulations for field applications of methyl bromide which could significantly reduce its use. Pesticide Action Network North America www.panna.org Canada consults on marine programme A National Action Programme for Protection of the Marine Environment is being set up by the Canadian government. Launching the Programme Fisheries and Oceans Minister David Anderson said that it was part of �a concerted eVort by Canada to take a radically diVerent approach to preserve and protect the oceans�one based on sustainability.� The draft NPA was prepared through the collaborative eVort of the federal provincial and territorial governments and was co-led by Environment Canada and Fisheries and Oceans Canada.It proposes national and regional programmes to protect the marine environment from land-based activities while recognising important variations between the Arctic Pacific St Lawrence and Atlantic regions. Environment Canada www.doe.ca/nat_action/index_e.html Germany proposes smog plan A draft plan to impose stricter controls on ozone pollution and so reduce summer smogs has been advanced by German Environment Minister Ju�rgen Trittin. One of the main measures is a cut in the emergency threshold for ozone from 240 to 180 mg m-3. This could lead to emergency actions being triggered on between 10�20 days each summer.The most controversial measure for Germans will be the proposal to introduce speed limits on motorways� German autobahns currently have no speed restrictions. The ministry claims that speed limits on motorways and other roads could cut ozone precursor chemicals significantly nitrogen oxides (NOx) by 10% and volatile organic compounds by 1%. German EnvironmentMinistry www.bmu.de Asia backs POP reduction Representatives from Asian and Pacific countries have endorsed moves by the United Nations Environment Programme (UNEP) to reach agreement on a global treaty to manage persistent organic pollutants (POPs) [see JEM 1(1) 10N]. At a recent meeting in Hanoi Vietnam delegates examined progress in moving toward a legally binding global agreement and examined the specific implications for the region.These include applying integrated pest management managing unwanted stockpiles of pesticides identifying and addressing releases of dioxins and furans and pursing alternatives to DDT for malaria control. The workshop was one of a series of regional meetings around the world in advance of the next round of treaty negotiations in Geneva in September. UNEP www.chem.unep.ch/pops/ Emission controls for landfill gas A series of measures to reduce the environmental impact of landfill gas are being proposed by the UK Environment Agency. The proposals include the 40N J. Environ. Monit. 1999 1 News ImagesA copyright 1999 PhotoDisc Inc. phasing out of open flares by the end of 2003 and the introduction of emission standards and monitoring standards for landfill gas flares.Landfill gas currently accounts for around 50% of total UK methane emissions. The new guidance on Best Practice Flaring of Landfill Gas forms part of the Agency�s strategy to improve local and global air quality. It will also help ensure the UK�s compliance with the EU�s proposed Landfill Directive. Final guidance will be issued later this year following a consultation period. UK Environment Agency www.environment-agency.gov.uk Detergents need to clean-up The environmental impacts of detergents are coming under increasing scrutiny in Europe from environmental NGOs and regulators. In April the Swiss Environment Agency said it would consider a ban on all use of the detergent surfactant nonyl phenol ethoxylate (NPE) unless current negotiations with industry lead to a voluntary phase-out.This and other alkyl phenol ethoxylates (APEs) have been linked to hormone disruption in humans and wildlife. Although other European countries have restricted the use of NPE in certain applications Switzerland is the first country considering an outright ban. The warning came with the publication of a new report from the agency on hormone disrupting chemicals in the environment. Meanwhile Brussels-based NGO European Environmental Bureau (EEB) has criticised the detergent industry�s environmental record. Pointing to experience in Sweden it says reductions in packaging and overall detergent consumption have been much higher than required elsewhere in Europe under a recent voluntary agreement between the industry and the European Commission.Between 1988 and 1996 cuts in the use of poorly biodegradable organic ingredients were achieved ranging from 15% for silicone defoamers to 50% for phosphonate and 100% for pigments fluorescent whitening agents and the complexing agent EDTA. EEB attributes the shift in Swedish consumers� buying patterns to eVective use of ecolabelling. Swiss Environment Agency www.admin.ch/buwal; Report ref �Endocrine Disrupting Chemicals in the Environment�; EEB www.eeb.org Denmark opts for lead ban A virtual ban on all uses of lead is being advocated in Denmark under a proposal currently being scrutinised by the European Commission and EU member states. Under the regulation products containing over 50 parts per million of lead would be banned from 1st November this year with prohibition eVective a few years later for various specific product groups.A wide variety of products would be aVected including major applications such as PVC plastic lead roofing materials ceramic glazes and crystal glass. Componentindustries would also be aVected. The European lead industry is protesting strongly against the move which it sees as leading towards EU-wide restrictions in the use of leadbased products. Danish EnvironmentMinistry www.mem.dk; L ead Development Association International www.ldaint.org Swedes to extinguish flame retardants A national ban on the sale and use of brominated flame retardants is likely to be introduced in Sweden following a critical report from the National Chemicals Inspectorate (Kemi). The report notes that levels of PBDE (polybrominated dephenylether) and PBB (polybrominated biphenyl) in human breast milk have doubled since 1992.The substances are widely used in television sets computers and cables but many of the companies questioned were unaware whether their products contained flame retardants or not. Reacting to the report Environment Minister Kjell Larsson said that since voluntary restrictions introduced in 1990 had been only partially successful a ban was likely. Swedish National Chemicals Inspectorate www.kemi.se Environmental quality Challenges for Central Europe Ten years after opening up to the market system the countries of Central and Eastern Europe still face major hurdles in improving environmental quality say Western agencies. In a report issued in March the Finnish EnvironmentMinistry detailed alarming environmental problems in the Karelian Republic the region of Russia bordering the whole of Finland�s eastern frontier.The Finns say practices such as deforestation peatland drainage and industrial pollution have severely damaged the region�s ecology.Many of the lakes and rivers commonly used for drinking water are contaminated by eZuent from the metal processing pulp J. Environ. Monit. 1999 1 41N News and paper industries. Raw sewage is regularly released into lakes and rivers near unprotected drinking water intakes causing repeated outbreaks of dysentery hepatitis-A and other gastrointestinal complaints. Too little has been done since the end of the Soviet era the ministry says. The Finnish-based Helsinki Commission an intergovernmental body monitoring the Baltic Sea also recently pointed to the continuing threat from pesticide practices in Russia and neighbouring countries.Another recent report from the region paints a more optimistic picture however. In a detailed review of Czech environmental performance published in April the OECD says that much progress has been made but that some emissions remain among the worst in any OECD country. The report commends the dramatic reductions in air emissions and the radical improvement in sewage treatment facilities. Nevertheless water quality �remains serious� and waste management also presents a major problem. Environmental expenditure has increased to over 3% of GDP over the last decade making it amongst the highest of any OECD country. Environmental policies have been one of the key areas in the Czech Republic�s negotiations to join the European Union.They also feature prominently in the EU�s negotiations with other applicant countries from Central and Eastern Europe such as Poland Hungary Estonia and Latvia. Finnish EnvironmentMinistry www.vyh.fi; Helsinki Commission www.helcom.fi; OECD www.oecd.org �OECD Environmental Performance Reviews Czech Republic� Asia sets up acid monitoring network Proposals for an Acid Deposition Monitoring Network in East Asia (EANET) have moved a stage nearer following an inter-governmental meeting in March. The network aims to create a common understanding of the state of acid deposition in East Asia and to provide useful inputs to decision-making on abatement policies. Participants in EANET at present include Indonesia Japan Malaysia Mongolia the Philippines the Russian Federation South Korea Thailand and Vietnam.China has been the most recent signatory. The Network is currently in a preparatory phase. An Interim Network Center has been established in Niigata Japan and each participating country has a National Center. An Interim Scientific Advisory Group is overseeing various technical issues such as the preparation of technical manuals quality assurance/ quality control programmes data reporting procedures and formats and training of personnel. In a further recognition of its pollution problems the Philippines is on the verge of passing its first clean air legislation. According to the World Health Organisation (WHO) airborne lead levels in Manila are more than three times the established safety limit and concentrations of suspended particulate matter are also dangerously high.The priority legislation will phase out the use of leaded fuel within 18 months reduce industrial emissions promote recycling ban incineration and drastically increase fines for owners of polluting vehicles. Observers say the real test of this ambitious programme will be eVective enforcement. Japan Environment Quarterly April 1999; BritishMedical Journal vol. 318 p. 689 Salmon hit by mystery disease Fewer Baltic salmon are now dying from the puzzling M74 syndrome say Swedish scientists but salmon deaths could increase again. Despite intensive studies it is still not clear what causes the syndrome which aVects newly hatched salmon fry. Researchers strongly suspect that the explanation lies in a number of complex interacting factors in the Baltic Sea.The condition usually results in the death of aZicted salmon fry within a few days of the first symptoms being seen. In 1993 when the frequency of M74 was at its peak the syndrome claimed up to 90% of farmed salmon fry in some rivers. In recent years the incidence has been considerably lower. Researchers have long suspected a link with environmental pollutants such as chlorinated organic compounds such as PCBs and dioxins but no link has ever been confirmed. Nor has evidence been found associating M74 with heavy metals bacteria or viruses. �It is becoming increasingly obvious that M74 is a complex environmental problem that demands extensive interdisciplinary co-operation� according to Prof. Bengt-Erik Bengtsson leader of a research project at Stockholm University.Precautionary treatment with vitamin B1 reduces mortality in salmon hatcheries but oVers no cure for wild salmon. Swedish EPA www.environ.se/fire Thinner ozone layer for 1999 The ozone layer over the northern hemisphere will be 1�5% less than �normal� this year according to scientists at Environment Canada. Normal values are defined by pre-1980 averages as ozone depletion was insignificant before 1980. The ozone layer was particularly depleted in 1993 when it was recorded at 7.2% less than normal. Last year was unusual as there was only minimal ozone depletion (0.2%) due to atypical weather conditions possibly related to El Nin�o. This year�s predicted ozone loss is typical of values observed in the 1990s except for 1993 and 1998. Canadian scientists say the 1�5% predicted ozone decrease corresponds to an increase of 1�6%in the average UV radiation although daily values can vary considerably.Environment Canada www.doe.ca ImagesA copyright 1999 PhotoDisc Inc. 42N J. Environ. Monit. 1999 1 News Pressure grows on phthalates The debate over the future of phthalate plasticizers has intensified following new concerns over their safety in both Europe and North America. Frustrated by a lack of action by the European Commission eight European countries have recently adopted unilateral restrictions on the use of certain phthalates in children�s toys. Austria and Denmark already ban the use of six phthalates in children�s toys and Germany Greece Italy Finland Sweden and non-EU member Norway are progressing towards national restrictions.Most of the national measures are similar and are framed in terms of banning the sale and production of PVC toys containing any of six types of phthalates that are intended or likely to be placed in the mouth of children up to three years old. Phthalates are used as softeners in toys and other PVC products. The European moves reflect concern about the migration of the chemicals from childcare items especially ones exposed to babies� saliva. These unilateral actions fly in the face of the opean Council for Plasticisers and Intermediates (ECPI) a body representing phthalate manufacturers. ECPI says the actions ignore advice from the EU�s own scientific experts the Committee on Toxicology Ecotoxicology and the Environment (CSTEE) which last year recommended safety limits on phthalate migration from PVC products.In any case the manufacturers say recent research shows phthalates to be safer than assumed by CSTEE. Studies now show much increased margins of safety for both DEHP and DINP the two main substances according to ECPI. The industry is particularly concerned by the erosion of confidence that could result from this piecemeal approach and has called for a Europe-wide solution. The eventual outcome of the toys issue is likely to have huge implications not just for other phthalate applications but for the whole chemicals agenda. Meanwhile in the US the new Center for the Evaluation of Risks to Human Reproduction has started a review of the possible risks from phthalates to plastics workers and consumers.The Center is Chemical hazards assembling a panel of a dozen or more scientists with applicable expertise to assess the health risks from seven phthalates commonly used in consumer products. (Animal data suggest the possibility of liver toxicity and reproductive eVects.) According to Dr.Michael Shelby of the National Institute for Environmental Health Sciences which sponsors the Center the panel will �look at data to see how likely it may be that phthalates contribute to reproductive and developmental problems�. It will also point to gaps in knowledge that need to be filled by additional research and testing. In contrast to the closed-door committees driving policy in Europe all sessions will be held in public. European Commission europa.eu.int/ comm/dg24; ECPI www.ecpi.org; NIEHS www.niehs.nih.gov Worldwide progress on chemical assessment A series of initiatives to help fill in gaps in basic health and safety information for mass produced chemicals have reached the operational stage.The International Council of Chemical Associations (ICCA) has finalised the initial list of chemicals under its High Production Volume (HPV) Initiative. The list prioritises chemicals that have either �a high volume of production in at least two countries or geographic regions� or are of concern because of their �wide dispersive use or potential for extensive human exposure�. Companies are being encouraged to work through ICCA to form consortia for collecting relevant information. The ICCA programme aims to provide complete toxicological and ecotoxicological data for 1000 HPV chemicals by 2004.In the US EPA has announced that 190 companies have committed to help provide data on HPVs under the Chemical Right-to-Know (CRTK) initiative. The HPV Challenge Program is a voluntary eVort to establish basic screening-level toxicity information on all 2800 chemicals manufactured in or imported into the United States in excess of one million pounds per year. Around 1100 chemicals will be covered in the first round. EPA is encouraging industry to voluntarily make available existing data and to conduct tests where there are gaps in baseline knowledge. Participating companies provide information on acute toxicity the eVects of repeated exposures reproductive and development toxicity and environmental eVects. A full list of chemicals guidance documents and other information is available at the CRTK website.ICCA www.icca.org; EPA CRT K www.epa.gov/chemrtk ImagesA copyright 1999 PhotoDisc Inc. Canada targets chemical toxics Stringent new measures to reduce industrial releases of certain toxic substances have been announced by Environment Canada. The plans include new prevention and control measures for the steel base metal smelting and metal finishing industries. Regulations will also be implemented for two specific toxic substances�dichloromethane and hexavalent chromium. Announcing the new measures Environment Minister Christine Stewart said the government was prepared to do whatever was �eVective and eYcient to get results on toxic substances�. If the voluntary programs failed to reach their targets she would �introduce regulations to achieve the expected results�.Releases of dichloromethane (DCM) will be reduced by 86% from 1995 levels by 2007. The sectors most aVected are J. Environ. Monit. 1999 1 43N News aircraft paint stripping flexible polyurethane foam pharmaceuticals adhesives and cleaning applications. The metal finishing industry will reduce releases of hexavalent chromium by 75% under a regulation which will also see significant reductions in cadmium and nickel. Toxic substances targeted for control in the steel industry include benzene PAHs lead mercury other metals dioxins and furans. The base metal smelting industry will target lead mercury arsenic compounds and nickel compounds with expected reductions of up to 90%. Environment Canada www.doe.ca Cancer warnings for creosote and bisphenol Concerns over the safety of creosote a substance commonly used as a wood preservative have been raised by the EU�s scientific advisory committee on toxicity issues.At a meeting inMarch the committee discussed the first ever full carcinogenicity study of creosote undertaken by Germany�s Fraunhofer Institute. Creosote contains trace amounts of a PAH called benzo-[a]-pyrene (BaP) but the product itself is classed as noncarcinogenic under EU law. The committee concluded that the German study provided evidence to �support the opinion that there is a cancer risk to consumers from creosote containing less than 50 ppm BaP and/or from wood treated with creosote�. The new findings will strengthen the case of four countries�Denmark Germany the Netherlands and Sweden�who are seeking national restrictions in creosote use and may even lead to EU-wide action.Another EU scientific committee has recently expressed concern over a chemical used in food container coatings. The opinion concerns bisphenol A diglycidyl ether (BADGE) a plasticiser capable of migrating into food and a known hormone mimic. Focusing on its carcinogenic properties the committee concludes that while there is no evidence for harmful eVects further research is needed to determine acceptable daily intake levels. It also repeats concerns raised in a 1997 report about chlorohydrins BADGE derivatives formed particularly in salty foods which are chemically very similar to known genotoxic substances. European Commission Health Directorate http://europa.eu.int/ comm/dg24 Public and occupational health WHO pushes for tobacco treaty The World Health Organisation has called on tobacco growers to back its eVorts to establish the world�s first public health treaty.The proposed Framework Convention on Tobacco Control would deal with a range of tobacco-related issues but would also ensure that individual countries are protected from the impact of multinational tobacco companies. WHO Director-General Dr. Gro Harlem Brundtland has identified global tobacco control as one of her major priorities. Tobacco growers must separate their concern for their livelihood from the public health impact of tobacco WHO representatives told a recent meeting of the International Tobacco Growers Association. However ITGA President Richard Tate said tobacco growers worldwide but especially in the developing countries were concerned about WHO�s initiative.He added that their concerns should not be confused with those of tobacco multinationals and urged WHO to keep farmers firmly in focus during the negotiations. WHO www.who.org French chart air pollution deaths The eVect of short-term air pollution excursions on mortality rates has been quantified in a study of nine French cities. The French Institute for Health Monitoring INSERM says air pollution accounts for 265 premature deaths each year. It adds that mortality rates increase by between 3�4% for a 50 mg l-1 rise in concentration of any of the four pollutants studied� particulates sulfur dioxide nitrogen dioxide and ozone. Cardiovascular deaths increase by between 2�5% and respiratory deaths by 1�6%. No major diVerences were found between gity rate in the French study contrasts sharply with an estimate of 24 000 premature deaths from shortterm pollution episodes attributed in a UK report last year. INSERM www.inserm.fr Drug cures aflatoxin US and Chinese scientists have successfully used the drug Oltipraz as an antidote to aflatoxin one of nature�s deadliest toxins. Writing in the Journal of the National Cancer Institute (vol. 91 no. 4) the scientists say the treatment ImagesA copyright 1999 PhotoDisc Inc. 44N J. Environ. Monit. 1999 1 News appears to have enhanced people�s ability to detoxify the natural agent and to have greatly reduced the amount of the chemical circulating in their bodies. Aflatoxin occurs on moldy grains Chemical industry looks to the long-run The chemical industry has launched a new global research initiative to explore long-term health and environmental issues associated with chemicals.The Long-range Research Initiative (LRI) is being jointly funded by the European Chemical Industry Council (CEFIC) the US Chemical Manufacturers Association (CMA) and the Japan Chemical Industry Association (JCIA). The budget is around US$25 million per year for at least five years. CEFIC says the initiative will help the industry �regain public trust enhance its reputation and defend its business from a position of strength�. It will also help inform �rational policy decisions�. LRI is being conducted in collaboration with academia and government. Its findings will be peer-reviewed published and made available to all interested parties.The industry says it is committed to undertaking timely action on the results in conjunction with other chemicals management initiatives. Priorities are initially in six areas environmental and human exposure assessment; risk assessment methodologies; chemical carcinogenesis; endocrine disruption; immunotoxicity and allergy; and respiratory toxicity. Attention will also be given to ecosystem dynamics; neurotoxicity; and atmospheric chemistry. In Europe the programme is being managed by the European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC). CMA will work closely with the Chemical Industry Institute of Toxicology (CIIT). L RI www.cefic.be/lri/lri.htm UK looks at scientific policymaking and green government The UK�s Health & Safety Executive (HSE) has announced a new research project to identify good practice and prepare guidelines for government when obtaining expert scientific advice.The Research news corn peanuts and other crops. Exposure can cause liver cancer. In many African and Asian countries large numbers of people regularly consume foods heavily contaminated with aflatoxin. Dr John Groopman of John Hopkins University research aims to identify principles of good practice for government departments in engaging scientific experts. Launching the project HSE Chief Scientist Dr. Jim McQuaid said the results will help ensure �consistency in the arrangements for securing scientific advice across government and for transparency in the way that advisers are selected�. The study is being undertaken by Oxford Economic Research Associates (OXERA) and will build on existing guidelines published in 1997.Areas covered will include the selection remit and independence of experts; the elicitation of their advice; and the use of advice in the wider decision-making process. On the same day another UK body the Environment Agency announced a research contract to help government departments and agencies set up their own environmental management systems (EMS). Under an initial twoyear arrangement government bodies are able to contact a central help desk to assist with the establishment of an EMS that meets the needs of their own business. The contract under the Greening Government Initiative was awarded to the consultancies ENTEC and W.S. Atkins. HSE toby_brown@oxera.co.uk; EMS www.environment-agency.gov.uk EA sets research priorities Improved methods and better information are the main thrusts of a new package of research measures announced by the UK Environment Agency.The Agency�s R&D programme for 1999/2000 aims to address both longterm strategic issues and shorter-term operational and policy needs. Around �11 million has been earmarked for the programme. Operational issues identified include the use of new technology for improving assessment of the state of the environment and priority areas in flood in Baltimore said the drug appeared to activate the P450 system part of the body�s natural defences against toxic poisoning. NIEHS www.niehs.nih.gov warning and forecasting. Strategic issues to be studied include impacts of climate change on the Agency�s regions and the development of a comprehensive strategy for managing chemicals.Most of the projects will be carried out through external contractors. UK Environment Agency www.environmentagency. gov.uk/envinfo/r_and_d/ RSC plans virtual conference The Royal Society of Chemistry together with the Royal Australian Chemical Institute will host an on-line conference later this year. Entitled T owards theMillennium�Modern Methods of Environmental Analysis the event will be held between 8th and 19th November. The Conference will comprise all the usual features of a scientific meeting� plenary lectures contributed papers discussion sessions and an exhibition. Presentations will cover relevant research analytical methods standards instrumentation data processing and interpretation. It will be the first time the RSC has held a conference of this type.Full details of the event including registration can be found at www.chemsoc.org or www.vei.co.uk Gas analysis network A network for scientists and business people concerned with gas analysis measurements was recently inaugurated by the UK�s National Physical Laboratory. NPL�s Analysis of Gas Awareness Club will keep its members informed on current developments in measurement techniques and requirements national and international standards and methods and other topics of interest. NPL Centre for Optical and Environmental Metrology paul.quincey@npl.co.uk NIEHS www.niehs.nih.gov J. Environ. Monit. 1999 1 45N News Chemical safety data The latest edition of a well known handbook summarises the most frequently used hazard health and environmental data for over 4500 of the most commonly used workplace chemicals.The data cover physical properties toxicity and health hazards and environmental considerations (based on US regulations). The new Genium Handbook is available as a three-volume set or on CD-ROM. Genium Handbook of Safety Health and Environmental Data Genium Publishing Corporation Schenectady. Workplace exposure guidelines The American Industrial Hygiene Association has published the 1999 edition of its Emergency Response Planning Guidelines andWorkplace Exposure Level Guide series. The 1999 ERPG Update Set contains 13 chemicals and an addendum for a Publications previously published ERPG; the 1999 WEEL Update Set covers 8 chemicals. Emergency Response Planning and Workplace Exposure Guidelines 1999 AIHA 1999 ISBN 0 932627 Safety now! Controlling chemical exposures at hazardous waste sites with realtime measurements AIHA has also published a self-help guide for health and safety professionals to assist in complex risk-assessments.The book focuses on the control of chemical hazards microbes noise radiation radionuclides and explosive vapours and covers over 600 materials. Includes a disk of sample spreadsheets. Hygienists guide The latest edition of the well known TLV/BEI Book is available from the American Conference of Government Industrial Hygienists (ACGIH). The book is a handy pocket-sized reference guide for recognising evaluating and controlling workplace exposures. More than 700 hazardous chemicals and physical agents are covered together with a list of substances and agents under study for the coming year.Details are on the ACGIH website at www.acgih.org Low wind dispersion A report from the UK�s Health & Safety Executive draws attention to the consequences of including low wind conditions in quantified risk assessments. The feasibility of alternative modelling approaches is also con9. Details from HSE Infoline tel:+44 (0)541 545500 ET �99. 8�10 June 1999 Birmingham. The UK�s largest environmental trade show. Details from Reed Exhibitions. E-mail et@reedexpo.co.uk 31st Mid-Atlantic Industrial and Hazardous Waste Conference. 20�23 June 1999 University of Connecticut Storrs CT. Details from Dr. Barth F. Smets e-mail bsmets@engr.uconn.edu or www.eng2.uconn.edu/environ 2nd National Conference on Natural Attenuation. 21�23 June 1999 SheYeld UK.The meeting will look at natural attenuation of organic pollutants in groundwater and soil. Details on-line at www.shef.ac.uk/~nnags/conf99/ 4th Annual Conference on Phytoremediation. 23�25 June 1999 Toronto Canada. Details from J. Gerson IBC Conferences e-mail jgerson@ibcusa.com or www.ibcusa.com/2280 Second Annual Environmental Forensics Conference. 24�25 June 1999 Washington D.C. USA. Details from J. Gerson IBC Conferences Events e-mail jgerson@ibcusa.com or www.ibcusa.com/2340 Water Environment Federation T echnical Conference. 27�30 June 1999 Indianapolis IN USA. Annual meeting covering all aspects of water monitoring and pollution control. Details from Eddie Gonzalez e-mail egonzalez@wef.org Society of Occupational Medicine Annual ScientificMeeting 1999. 14�16 July 1999 Plymouth UK.Conference covers the scientific and practical aspects of occupational medicine. E-mail sayeed.s.khan@rolls-royce.btx400.co.uk Health Consequences of Phytoestrogens in Foods. 15 July 1999 London UK. Details from H. Wiseman Society of Chemical Industry tel:+44 (0)181 654 8216 6th EuropeanMeeting onMass Spectrometry in Occupational and Environmental Health. 1�3 September 1999 Stockholm Sweden. Details from Prof. Anders Colmsjo� e-mail anders.colmsjo@anchem.su.se or www.anchem.su.se/emmsoeh6 Instrumental Methods of Analysis. 19�22 September 1999 Chalkidiki Greece. Details from IMA�99 Secretariat tel +30 1 7723098 or www.chemeng.ntua.gr/IMA99/IMA99.htm 1999 Chemical Emergency Preparedness and Prevention Conference. 20�23 September 1999,Washington D.C. USA. Conference organised by EPA focusing on the Emergency Planning and Community Right to Know Act.Details from www.epacepp.com Environmental Problem Solving with Geographic Information Systems A National Conference. 22�24 September 1999 Cincinnati OH USA. Organised by EPA�s National Risk Management Research Laboratory (NRMRL). Details from www.epa.gov/ttbnrmrl/ gisann.htm T he Role of Human Exposure Assessment in the Prevention of Environmental Disease. 22�24 September 1999 Rockville MD USA. A workshop organised as part of the National Toxicology Program�s work on human exposure assessment. Details from NTP Liaison and Scientific Review OYce tel +1 919 541 0530 46N J. Environ. Monit. 1999 1 News Environment Japan �99 T he 16th JETRO Import Fair. 20�23 October 1999 Osaka Japan. An international trade fair to be held alongside the New Earth �99 Conference.Details from www.environmentjapan.org 9th International Emission Inventory Conference Regional Strategies for the Future. 25�28 October 1999 Raleigh North Carolina USA. Details from Sally Dombrowski EPA tel +1 919 541 3269 Measurement for success UK National Measurement Conference �99. 2�4 November 1999 Brighton UK. Incorporates BEMC �99�The 9th International Conference on Electromagnetic Measurement. Details from NMP Conference Secretariat National Physical Laboratory. e-mail nmp_sec@npl.co.uk 1999 Eastern Analytical Symposium. 14�19 November 1999 Somerset NJ USA. Details from EAS Program Committee e-mail easinfo@aol.com Wetlands and Remediation An International Conference. 16�17 November 1999 Salt Lake City UT USA. Scope will include both the treatment and remediation of contaminated wetlands and the use of wetlands for the treatment and remediation of contaminated water and wastewater.Details from Karl Nehring Battelle Memorial Institute e-mail nehringk@battelle.org Food Hygiene �99. 24�25 November 1999 Telford UK. Exhibition and conference covering food hygiene monitoring control implementation and training. Details from Fiona Angus Leatherhead Food RA e-mail fangus@lfra.co.uk Fourth Workshop on Biosensors and Biological T echniques in Environmental Analysis. 1�3 December 1999 Menorca Spain. Organised by the International Association of Environmental Analytical Chemistry and the Institut Menorqu� d�Estudis. Details from dbcqam@cid.csic.es FDA and the Science of Safety New Perspectives. T he 2000 FDA Science Forum.14�15 February 2000 Washington D.C. USA. Conference on the role of science in risk assessment organised by the US Food & Drug Administration. Details at www.fda.gov/oc/meetings/ science2000.html 30th International Symposium on Environmental Analytical Chemistry. 13�16 June 2000 Espoo Helsinki Finland. Details from International Association of Environmental Analytical Chemistry e-mail iaeacmfrei@access.ch or tiina.harju@vtt.fi 26th International Congress on Occupational Health. 27 Aug�1 Sept 2000 Singapore. The Annual Meeting of the International Commission on Occupational Health (ICOH). Details from ICOH2000 e-mail icoh2000@post1.com or www.icoh.org.sg X2001 Conference on Exposure Assessment in Epidemiology and Practice. 10�13 June 2001 Go� teborg Sweden. Details from x2001@ymk.gu.se or www.ymk.gu.se A New Era of Occupational Hygiene T he 5th IOHA International Scientific Conference.10�14 June 2002 Bergen Norway. Details from bjorg.hollund@isf.uib.no US regs on CD-ROM All 24 volumes of the so-called �Title 40��EPA�s Environmental Regulations�of the US Code of Federal Regulations are available on CD-ROM. The latest oVering updates the regs to the 1998 revision levels. It features 5000+ in-line graphics and instant search and retrieval in PDF file format. Updated information on chemicals regulated under EPA�s Toxic Substance Control Act (TSCA) has also been published. A new CD-ROM lists 62 000 chemicals cross-referenced with their SARA Title III reporting requirements. The data is also cross-referenced to the EPA PMN Canadian DSL/NDSL and European ELINCS conventions.A third CD-ROM set presents a database of over 225 000 Material Safety Data Sheets. The set features the Environmental Reporting Assist File and Registry of Lists databases. Details on all three products are available from www.env-sol.com/ solutions/ UK site data A new commercial service called Sitescope oVers extensive property and Info gateways environmental data for sites in England andWales. Aspects covered include abstraction licences chemical release inventories discharge consents hazardous sites landfill sites to name a few. The data can be searched either by postcode or by national grid reference to determine particular environmental risks on or around any site and is available on CD-ROM. Catalytic Data L td www.sitescope.co.uk Canada expands OHS data The Canadian Centre for Occupational Health and Safety (CCOHS) is expanding its CCINFOweb Internet service.This now includes the International Labour Organisation�s CISILO database covering occupational health and safety literature worldwide. In addition CCINFOweb now includes the NIOSHTIC database from the US National Institute for Occupational Safety and Health and HSELINE from the UK�s HSE Information Service. The subscription based service now runs to over 400 000 records. CCOHS www.ccohs.org German Internet data Air and water quality monitoring data from around Germany are available on a new website. Aimed mainly at members of the public the site covers four air pollutants�sulfur dioxide nitrogen oxides ozone and particulates�and 40 water-borne pollutants from various monitoring points around the country.The site is at www.umweltbundesamt.de/uba-infodaten/ daten/schadsto Biotech means business Scientific publisher STN International has added the BIOCOMMERCE database to its on-line oVering. BIOCOMMERCE covers business aspects of biotechnology and the biological sciences worldwide. Information contained in the database includes company news and financial data product news research results patents and technology licensing. STN is also oVering FROSTI a database on food sci operated by EPA which have seen major updates recently (in content rather than presentation) include Alternative Fuels www.epa.gov/oms/consumer/fuels/ altfuels/altfuels.htm Great Lakes Programme www.epa.gov/glnpo National Nutrient Criteria Development www.epa.gov/ost/standards/nutrient.html Superfund Reforms www.epa.gov/superfund/programs/ reforms.index.htm Watershed Information Network www.epa.gov/win Laboratory.com Laboratory Network is an online community for the analytical sciences.The site is aimed at chemists researchers laboratory managers and research directors. Coverage includes daily news updates and reports on business and technology trends within the industry. Surfers can sign up for a free subscription to a weekly newsletter which includes news and product reviews (mainly related to the US market). If you like what you see you can also purchase laboratory equipment and supplies. The site is at www.laboratorynetwork.com Cyber Hospital A major new resource for health professionals has been launched in the UK.Cyber-Hospital aims to provide one-stop access to medical information for doctors and associated professionals in the medical industry. Services on oVer include the latest medical news a library of research findings from around the world careers advice and training resources. Involvement is being sought from commercial interests as well as the medical world. www.cyber-hospital.org.uk Online chemistry The Royal Society of Chemistry has launched an online periodic table as part of a collaborative project in science and the visual arts. The online table features pictorial interpretations of all the elements together with brief descriptions of their properties and uses. A selection of desktop patterns screens and screensavers is available for download.Details at www.chemsoc.org/viselements Distance learning A new service oVers scientists access to work-based training through use of distance learning technologies. Science Learning Center is a career development tool that aims to teach scientists workplace skills. www.sciencelearning.com Newsletters Latest editions of some occasional newsletters accessible on-line Pesticides News No. 43 March 1999. The Journal of the Pesticides Trust with articles on persistent organic pollutants genetically modified organisms residues in food pesticide issues in developing countries and use of pesticides on aircraft. Plus book reviews and on-line sources. www.gn.apc.org/ pesticidestrust/pn43.htm Environmental Health & Safety News No. 8 April 1999. An occasional newsletter from OECD with updates on OECD�s work on testing guidelines endocrine disruptors good laboratory practice chemicals management risk assessment the High Production Volume Chemicals programme pesticides chemical accidents and biotechnology.www.oecd.org/ehs Science and the Environment Bulletin No. 11March/April 1999. Environment Canada�s on-line newsletter with articles on phytoremediation mercury pollution in the Arctic bio-markers as indicators of chemical pollution air contamination in the St Lawrence river valley and water quality in the Third World. www.doe.ca EPA Newsletters Latest issues of numerous newsletters issued by EPA agencies covering virtually all aspects of EPA�s work can be accessed from the EPA Newsletters homepage www.epa.gov/epahome/newslett.htm. Many are available through free e-mail subscription. J. Environ. Monit. 1999 1 47

ISSN:0960-7919    

DOI:10.1039/a903472b

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

Forum Summary of environment-related research areas developed in the frame of the 5th Framework Programme of the European Commission Thematic programmes The four Thematic Programmes constitute the First Activity of FP5 and are aiming respectively to (1) improve the quality of life and management of living resources; (2) create a user-friendly information society; (3) promote competitive and sustainable growth; and (4) preserve the ecosystem. Each Thematic Programme is organised in the form of Key Actions Generic Technologies and Research Infrastructures. Each Key Action is problem-oriented and targeted to the objectives of the respective Programme. The aim is to link them to major and social objectives with relevant national and international initiatives and to integrate activities and disciplines (from basic research to development and demonstration) in close consultation with the scientific community the industry and the users.Generic Technologies aims to develop the European technological capacity and to stimulate a flow of ideas and knowledge to complement the Key Actions. They focus on a limited number of areas not covered by the Key Actions and on potential multi-sectoral applications. The Research Infrastructure activities promote an optimal use and wide access to existing research infrastructures and develop transnational cooperation in a rational and cost eVective context. This Community added value is complementary to national and multinational initiatives (e.g. networking). Horizontal programmes These correspond to the Second Third and Fourth Activities of FP5 which are dealing with (1) International Role of Community Research; (2) Innovation and Participation of SMEs; and (3) Human Potential and Socio-economic Research.These activities are based on coordination support and accompanying measures of Thematic Programmes and support other Community policies. A brief description of environmentrelated activities covered by two thematic programmes is given below. Some information is also given on horizontal programmes. 2. Environment in the programme on �Quality of life and management of living resources� The programme focuses on immediate socio-economic and market needs in areas such as improving our food controlling disease harnessing the power of the biological cell the sustainable development of agriculture and fisheries and a healthy and independent old age.One of the key actions deals specifically with environment as described below. Full details on this action are accessible via the Internet (see address above) and through the programme help-desk life@dg12.cec.be Key Action 4 �Environment and health The following priorities will be considered (1) diseases and allergies related to or influenced by the environment their prevention and treatment; and (2) development of new methods of diagnosis risk assessment and processes to reduce causes and harmful environmental health eVects. 48N J. Environ. Monit. 1999 1 Following the decision of the European Parliament and of the Council of 22 December 1998 The European Commission has launched its Fifth Framework Programme for Research Technological Development and Demonstration (RTD) which will run from 1999 to 2002.The activities will be carried out in the frame of four thematic programmes and three horizontal ones. The first calls for proposals were published in March 1999. Detailed information on the work programme of each activity is available through the Internet at the following address http://www.cordis.lu/fp5 and through direct contact with the programme�s helpdesks. The present summary gives an outline of environment-related research activities covered by 5th FWP. 1. Introduction The Fifth Framework Programme (1999� 2002) is simply designed to support and enhance scientific and technological excellence while being relevant to main EU policies and to European added value. The objectives are defined according to socio-economic and competitive needs placing more emphasis on the results and focusing on a limited number of topics.In comparison to the Fourth Framework Programme (1994� 1998) a simplified structure is proposed consisting of four Thematic Programmes and three Horizontal Programmes Thematic Programmes Quality of life and User-friendly Competitive and Preserving the living resources information society sustainable growth ecosystem Programme 1 Programme 2 Programme 3 Programme 4 Key Actions+Generic Technologies+ Research Infrastructure Horizontal Programmes International Role of Innovation and Human Potential & Community Research Participation of SMEs Socio-Economic Research Coordination and Specific Actions 3. Environment in the programme on �Competitive and sustainable growth� Activities developed under this programme should help to prepare the policy making industrial and related service sectors for the challenges of the new millennium.Environment-related activities are mainly focusing on material research (e.g. waste recycling) and reference materials in the frame of two generic activities described below. Additional information on the programme can be obtained at the following address growth@dg12.cec.be Generic Activity 1 �Materials and their technologies for production and transformation This activity covers four main objectives (1) cross-cutting generic materials technologies; (2) advanced functional materials; (3) sustainable chemistry; and (4) expanding the limits and durability of structural materials. Generic Activity 3 �Measurements and testing The RTD priorities are (1) instrumentation; (2) methodologies for measurements and testing; and (3) support to the development of Certified Reference Materials (CRMs).It should be noted that environment-related research is not covered by the two first items (which are covered in the programme 4 described below) whereas the development of environmental CRMs is covered by the activity. 4. The programme �Preserving the ecosystem� This programme is the best suited platform for submitting environmentrelated projects it is centred around two main parts namely �Environment and Sustainable Development� (Part A) and �Energy� (Part B). Only those actions relevant to environmental monitoring are described here. Additional information can be obtained through the programme help-desk at eecs@dg12.cec.mail Key Action 1 � Sustainable management and quality of water This key action covers several items such as (1) water management in cities; (2) waste water treatment and re-use prevention of diVuse pollution; (3) prevention and abatement of water pollution from contaminated land landfills and sediments; (4) integrated water resources and wetland management at watershed scale; and (5) management of water resources in water-stressed regions.Key Action 2 �Global change climate and biodiversity This key action focuses on several items such as (1) improving the knowledge of global change climate and biodiversity; (2) analysing the impacts; and (3) responding to the global issues. Key Action 3 �Sustainable marine ecosystems In this key action the following activities will be considered (1) marine processes ecosystems and interactions; (2) monitoring and managing coastal processes and the coastal zone; and (3) operational forecasting in coastal shelf and slope areas.5. Horizontal programmes As mentioned in the Introduction horizontal actions are developed by three programmes in close coordination with the thematic programmes. In this frame environmental-related research may be envisaged for example in the form of projects in partnership with Third Countries organisations (programme �International RTD co-operation� E-mail inco@dg12.cec.be) projects aiming to encourage and facilitate the participation of small and medium enterprises in RTD activities (programme on �Innovation and the participation of SMEs�) and projects for improving human potential through training and mobility (programme �Improving the human research potential and the socio-economic knowledge base� E-mail improving@dg12.cec.be).6. Conclusions Implementation of RTD activities is generally carried out through calls for proposxed dates) related to key actions open calls (e.g. for SME specific measures fellowships and accompanying measures). The modalities for the diVerent actions are described in detail in the work programmes of the diVerent Thematic and Horizontal Programmes. Preparing a proposal to be submitted for funding in one of the specific programmes of the 5th FWP requires a good reading of the respective work programmes. As summarised above each programme describes in detail the various types of activities in which RTD projects may be submitted.Proposers should not �reinvent the wheel� and stick closely to the areas described to enhance the chance of their proposals being selected. It should be noted that each proposal will be evaluated by three independent experts contracted by the Commission; the evaluators will judge the submitted projects in the light of the respective work programme and following well defined evaluation criteria. It is hence strongly recommended that proposers closely follow the instructions given in the guide for preparing proposals and consider the main evaluation points listed (Guide for Proposers Part 2); proposers should be able to respond to all questions expressed in the evaluation guidelines while reading their project proposal� in other words the evaluation guidelines should be considered as a �red line� for identifying the most critical points to the considered when finalising a proposal.As mentioned above general information on all Key/Generic activities and other activities is accessible via the Internet and the programme�s help-desk. Proposers may contact EC Scientific OYcers for specific questions; it should be noted however that contacts should be restricted to well formulated questions (which cannot find a response in the available documents) and not general ones; in other words please read the work programmes and related documents carefully before directly contacting EC personnel! Philippe Quevauviller Brussels Belgium J. Environ. Monit. 1999 1 49N Forum interdisciplinary fields contained within this scenario. For this the organisers brought together a strong international contingent of experts from nine countries.The attendees were drawn from a similarly wide international range. In addition to the formal scientific sessions there was a large poster exhibition containing about 40 new scientific contributions as well as an instrument exhibition both of which ran for the duration of the symposium. In addition to the scheduled events ample time was allowed for the 150 or so attendees to meet and talk informally. The net result was a very relaxed environment for the exchange of new ideas and information relevant to air quality measurement in all its contexts. Contributions on exposure assessment strategies identified the various motivations for exposure assessment placing air monitoring firmly within the need to measure the exposures of people to airborne contaminants via the inhalation route in a way that relates to outcome (e.g.to human health). So it was understood that air monitoring is an important component of environmental and occupational health in the wider context of public health. The needs of epidemiology were discussed most notably from the point of view that sampling strategies provide a primary scientific basis for the development of standards and exposure limits for airborne contaminants. The role of variability in exposure assessment was articulated where the importance of placing statistical considerations alongside the technical ones that have dominated the subject for so many years was stressed. One contribution developed the new argument that selfassessment of exposure might be useful in the future.Overall the new work which was presented suggested that the training of environmental and occupational hygienists and scientists needs to go beyond the technical disciplines physics chemistry and biology which are frequently regarded as the cornerstones of graduate-level education in those fields. Meanwhile the technical methods by which to carry out visualisation of exposures to aerosols gases and vapours were highlighted as providing an important body of practical tools by which hygienists in the workplace setting may explore the nature and exposures for individual workers (and groups of workers). The information gained in this way may be used to improve working practices and educate workers about how to reduce their exposure(s). The session on aerosols returned to themes developed at earlier AIRMON symposia; namely that aerosol exposures need to be measured in a way that physically presents the way in which people are exposed.The question of the role of external factors such as particle size windspeed (including moving versus calm air scenarios) and sampling flow rate were therefore addressed leading to suggestions for further updating the criteria which underpin current particle size-selective sampling and for the development of new�and more user-friendly� sampling instrumentation. The importance of micro-organisms as an important aerosol sub-group was identified their measurement diYculties were discussed and new measurement procedures were described. The special importance of very very small particles was outlined where it was noted that the very reactive properties of particles in the size range of just a few tens of 50N J.Environ. Monit. 1999 1 Forum AIRMON �99 The Third International Symposium on Modern Principles of Air Monitoring (Geilo Norway February 10th to 14th 1999) This symposium was the third in the series of such meetings the first being held at Geilo in 1993 and the second in Sa�len Sweden in 1996. This latest meeting marked the return to the location of the first and was organised jointly by the National Institute of Occupational Health of Norway (Oslo) and the National Institute for Working Life of Sweden (Umea� ). It provided an auspicious launching of the Royal Society of Chemistry�s new journal the Journal of Environmental Monitoring (JEM) in which key papers presented at this meeting will be published later this year.The rationale for the meeting was the increasing requirement for air monitoring within the preventive frameworks for identifying and controlling airborne health hazards in workplaces and in the ambient and living environments. These in turn demand the characterisation of exposures of people to chemical and biological agents involving the use of representative and valid sampling procedures and strategies in addition to accurate and reliable quantitation techniques. With this in mind the programme of plenary lectures was designed to provide comprehensive overviews of the latest developments in the multidisciplinary and nanometers which make them of great interest from an engineering point of view (e.g. new materials) are also the same ones that might make them very toxic.One type of aerosol exposure that continues to provide significant measurement challenges concerns that from machining fluid aerosols experienced in the many industries where metal working is carried out. In this context the special problems involved in measuring aerosol exposure associated with the use of the aqueous class of machining fluids were discussed. Finally the emergence of new techniques for the chemical analysis of individual particles was described and the importance of the application of such techniques in �finger-printing� aerosols in the working and ambient environments was stressed. The discussion about gases and vapours addressed concern about how air monitoring might deal with the diYcult practical question of the measurement of airborne contaminants that may exist in both the aerosol and the gaseous form.Isocyanates continue to be an important airborne contaminant in many industrial settings and still provide many measurement challenges. For these emerging new methods were summarised and compared with the ones that were used some 20 or so years ago. It was described how complex isocyanate mixtures add to the diYculties that are currently faced and how significant progress is being achieved. New methods for the sampling of inorganic gases and vaortance of aldehydes was also identified and a range of analytical methods for their determination was described. Biosensors appear to have very high sensitivity for the measurement of gases and vapours and it was suggested that these have great potential for measuring extremely low concentrations.Many air quality issues have emerged in recent years. We are finding that health eVects can occur at concentrations of airborne contaminants which compared to most workplace and ambient air quality standards are very low indeed. In this respect the importance of hydrocarbons ozone and other chemical species groups was discussed and new air monitoring challenges were identified. For many such situations where�unlike for the assessment of workplace exposures�the culture (and practicality) of personal sampling for the determination of individual exposures has not yet been fully developed dispersion modelling can provide important insights. In particular therefore as was described in one contribution such modelling can provide a means for determining the transport of airborne contaminants and hence for assessment of their temporal and spatial distributions in order to provide better estimates of the exposures of people in airsheds both large (i.e.the atmosphere) and small (i.e. indoor air situations). Indoor air quality remains of considerable interest especially since so-called �sick building syndrome� continues to be among the most commonly reported of workplace complaints. Here the role of microbegenerated volatile organic compounds (VOCs) were discussed. It is known that there is generally no relationship between SBS symptoms and levels of individual or total VOCs. However it was suggested that a multivariate statistical approach on the measurement of VOCs may provide a means for the classification of �problem� and �non-problem� buildings.Overall the development of more systematic scientific approaches to the understanding of this important problem was called for. The last part of the symposium was devoted to discussion of questions concerning international regulations standardisation and quality assurance. For most of the attendees the European Union (EU) framework was clearly of primary interest and this was addressed in relation to the new EU air quality directives methods standardisation reference procedures and proficiency testing schemes. The speakers described the various mechanisms that are in place or are being developed and the extent to which harmonisation across the EU is being achieved. It would appear that much but perhaps not all of what was described is generic and so applicable to countries outside the EU.The symposium followed in the same tradition as the first two once again reflecting the commitment of its two Co-chairs Yngvar Thomassen and Jan Olof Levin (of the mentioned Oslo and Umea� institutes respectively) who have organized and hosted all three symposia in the series. Once again the tone of the symposium owed much to the contributions of these two individuals. As before the symposium was notable not only for its excellent and broad scientific content but also for the congeniality of its organisation and surroundings. There are few scientific meetings most of us attend where the lunch break begins at noon and goes on until 4.00 pm yet where it just so happens that some of the world�s best ski slopes and trails beckon just outside the door! And where during the early evening sessions freshened by our exposure to the clean winter air of Geilo we could look ahead to the bountiful tables provided by the superb kitchens of the Dr.Holms Hotel! I think I can speak for everyone who attended in thanking our hosts for yet another outstanding air monitoring symposium. We all look forward to AIRMON �02. James H. Vincent University of Michigan Department of Environmental and Industrial Health School of Public Health Ann Arbor MI 48109 U.S.A. J. Environ. Monit. 1999 1 51N Forum The Waste Testing and Quality Assurance Symposium (WTQA�99) will hold its fifteenth annual meeting in Arlington VA this summer. The symposium is sponsored by the Waste Policy Institute (WPI) with the support of the EPA. The conference will be held 1999 WTQA Conference July 18�22 at the Crystal Gateway Marriott Hotel 1700 JeVerson Davis Highway Arlington VA 22202 (tel +1 800 228 9290).Meeting pre-registration discount rates ($295) are valid through June 15 1999. Hotel room discount rates ($120+tax single or double room) are valid through June 28 1999. The theme of WTQA�99 is �Preparing for Change Under PBMS.� Performance Based Measurement System (PBMS) is a new regulatory approach to compliance monitoring that has been initiated by EPA. Under the PBMS approach facilities may use any scientifically valid technology or method to demonstrate that they are in compliance with permit or other regulatory standards rather than only having to use EPA approved methods. Through this new approach the Agency expects to reduce the incurred by regulated entities to demonstrate regulatory compliance help laboratory to improve their productivity eliminate a major barrier to the development and use of innovative faster less costly measurement technologies and methods; and in some cases improve the quality of environmental monitoring data and compliance determinations.While PBMS will give the analyst freedom in how one carries out the required data gathering it also imposes a responsibility on the regulated industry (and its laboratories) to demonstrate that the methods used are yielding accurate compliance determinations. The environmental community faces many new issues with respect to PBMS. AVected parties include permit writers in the federal and state agencies state and federal enforcement oYcials regulated entities the environmental laboratory community and the various laboratory accreditation programs.WTQA�99 will provide attendees with the latest information from federal and state oYcials on their progress towards implementation of PBMS. Attendees of WTQA�99 will learn how EPA is implementing PBMS in its various regulatory programs and will be able to interact with oYcials who are developing the new regulations and program operation guidance. The three day conference will begin with a plenary session on July 19 followed by concurrent technical sessions on July 20�21 1999. The conference includes a Tabletop Exhibition and will be preceded and followed by short courses. In addition to regular technical sessions on quality assurance organic and inorganic chemical analysis six special PBMS Issue Sessions focus on PBMS Status Contracting Laboratory Management Scientific and Legal Defensibility Field and Laboratory Implementation and Laboratory Auditing and Accreditation.Detailed hotel and program information is available on the Internet at http://www.wpi.org/wtqa or by sending a fax to +1 540 557 6043 attention Dr. Larry Keith. The program for the two days of the main symposium is summarized below Tuesday July 20 1999 PBMS Status and Issues (1 2 day morning)�The latest information on draft rules and regulations from the RCRA CERCLA CWA SDWA and CAA programs plus reports from NELAC ELAB ACS and the interagency Methods and Data Comparability Board. Quality Assurance (1 2 day morning)� Presentations on reduction of costs by eliminating unnecessary QC samples streamlining EPA�s DQO process PBMS at the Rocky Mountain Arsenal PBMS case study with rapid response field analytical support and more.Environmental Business in the PBMS Paradigm (1 2 day afternoon)�Features sub-sessions on Contracting (including frameworks for contracting under PBMS changes in Superfund contracting and a model agreement) and Laboratory Management Issues (including managing labs under PBMS laboratory performance expectations consensus standards roles and more). Inorganic Analysis (1 2 day afternoon)� Presentations on EPA�s Inorganic Methods Program perchlorate analysis new developments with analysis of mercury tin lead zinc and copper new developments with Method 7473 and more). Organic Analysis Poster Session (all day)�Presentations on a wide diversitys and SVOCs with Methods 3511 and 3570 Method 8261 modifications analysis of chemical nerve agents groundwater headspace sampling in the field soil cleanup technique for Toxaphene new approach for simultaneous selected ion and full ion scanning MS PBMS round robin standards for petroleum hydrocarbons and chlorinated dioxin and furan IA field analysis and others.Wednesday July 21 1999 PBMS Implementation (1 2 day morning)�Features sub-sessions on Ensuring Scientific and Legal Defensibility (including an overview of EPA�s approach the Comparable Fuels Rule as a model and perspectives on quality assurance private laboratories legal and enforcement issues) and Field and Laboratory Implementation Issues (including how to develop DQOs developing project-specific MQOs moving MQOs into commercial laboratories and balancing error sources for project planning).Laboratory Auditing and Accreditation under PBMS (1 2 day afternoon)� Highlights new roles of auditing and accreditation laboratory compliance programs changes in state auditor�s roles documentation requirements community issues and expectations of government laboratories. Organic Analysis (full day) Presentations on EPA�s Organic Methods Program storage stability of VOCs in soil coste Vective soil sampling techniques recovery comparisons of VOCs from soils modifications of Methods 8100 for PAHs and 3545A for diesel range organics and waste oil organics new biosensors for environmental endocrine disruptors and analysis of carbamates and more. Quality Assurance Poster Session�(full day) Presentations include auditing waste characterization methodologies performance evaluation studies contaminated marine sediment reference material and current activities in standard reference materials for organic contaminants and others.Inorganic Analysis Poster Session�(full day) Presentations on a wide diversity of topics including a universal ICPOES method for environmental analyses new technologies for metal digestions ICP-MS under PBMS using acid mine drainage to detoxify hexavalent chromium leachate applications of in-situ gamma spectrometry with radioactively contaminated soil and more. WTQA �99 will be the place to learn about critical policy issues the latest developments in monitoring methods and technologies problems that have been encountered when using existing methods and the new approaches being developed for auditing laboratories and analytical data for compliance with EPA State and National Environmental Laboratory Accreditation Conference (NELAC) standards. The conference will bring together regulators analysts engineers and managers from Federal and State regulatory agencies the regulated community and the laboratory and engineering support communities. 52N J. Environ. Mon

ISSN:0960-7919    

DOI:10.1039/a903473k

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

Determination of metal–humic complexes, free metal ions and total concentrations in natural waters Petra K. Appelblad,ab Douglas C. Baxter*a and Jonas O. Thunbergc aDivision of Inorganic Chemistry, Lulea° University of Technology, S-971 87 Lulea° , Sweden. E-mail: douglas.baxter@km.luth.se bSGAB Analytica, Lulea° University of Technology, S-971 87 Lulea° , Sweden cDivision of Applied Geology, Lulea° University of Technology, S-971 87 Lulea° , Sweden Received 8th February 1999, Accepted 22nd March 1999 A comparison of two systems for the quantitation of metal–humic complexes and free metal ions, consisting of the separation by coupled ion exchange columns followed by detection by inductively coupled plasma mass spectrometry or cold vapour atomic fluorescence spectrometry, is presented.The systems evaluated comprised the serially coupled anion and cation exchangers, Sephadex A-25/Chelex 100 and Dowex 1X8/Chelamine Metalfix. Separation and preconcentration of the species studied were accomplished with both systems, elution being carried out using 2 MHNO3. Total concentrations, metal–humic complex fractions and free metal ion fractions of Al, Ba, Cd, Co, Cu, Fe, Hg, Mn, Pb, Sr, U and Zn in nine natural waters were determined.Statistical evaluation of the data from the two cation exchange materials, including results for additional elements, showed better precision (for Al, Ba, Cr, Cu and Mo) and higher recoveries (Al, Ba, Cd, Fe, Sr and Zn) for Chelex 100 than Chelamine Metalfix for free metal ions. On the other hand, Chelamine Metalfix recovered a significantly greater amount of Ni.The amounts of metal–humic complexes were compared with modelled distributions of these species, and one advantage of the preferred Sephadex A-25/Chelex 100 system is that the elements studied are all correctly classified with respect to their binding strengths to humic substances, which is not the case with the Dowex 1X8/Chelamine Metalfix pair.With the preferred system, metal–humic complexes can be reliably determined, as indicated by the results of equilibrium speciation modelling. However, comparison with the total concentrations showed statistically significant, non-quantitative recoveries of Al, Cu, Hg,Mn, U and Zn from some samples. Thus a combination of speciation and total concentration measurements is required to obtain a complete representation of the distribution of trace elements in natural waters.Humic substances are a general category of naturally occurring at least 94% of the latter neutral complexes were retained on a reversed phase non-polar C18 packing placed upstream of heterogeneous macromolecules, formed through the breakdown of plants and animal tissues by chemical and biological the Dowex 1X8 anion exchanger in a three-column system.The final column contained Chelamine Metalfix, in which the processes. They form both soluble and insoluble complexes with polyvalent cations, depending on the degree of saturation, chelating group, 1,4,7,10,13-pentaazatridecane, is attached to an organic polymer. This resin was investigated by Blain as is well documented by many experimental1–5 and modelling6 –8 studies.Complexation of trace metals by humic sub- et al.,12 who reported that Chelamine Metalfix allows the preconcentration of and matrix elimination for Cd, Cu, Ni, stances may aVect the aquatic system in two ways: by decreasing the toxicity of certain metals and by increasing the Pb and Zn over a broad pH range in saline waters without prior treatment such as buVering.The combination of C18, availability of others.9 KerndorV and Schnitzer10 reported, in one study, the relative strengths of the bonds to humic acid Dowex 1X8 and Chelamine Metalfix permitted the isolation of neutral complexes, anionic complexes and free metal ions, for metal ions. Depending on the stabilities of the bonds, the metal ions may be divided into three groups exhibiting strong, respectively, from fresh water samples.One disadvantage inherent in the use of strong ion intermediate or weak interactions with humic substances. The elements that were found to be most tenaciously bound by exchangers, such as Dowex 1X8, is irreversible sorption to the material. Kim et al.13 attributed this phenomenon to the humus were iron and mercury, while lead, copper and aluminium formed complexes of intermediate strength.Cobalt, high aYnity of phenolic compounds, which would include humic substances, for quaternary amine sites. An eYcient cadmium and zinc displayed weak interactions with humic substances. Chromatographic techniques,1,3–5,11 such as solid weakly basic resin is diethylaminoethylcellulose (DEAE Sephadex A-25), which has a macroporous structure and a phase extraction (SPE), have proven to be eVective experimental approaches to the isolation of humic substances.The hydrophilic matrix to which secondary amines are attached. This resin has been found to be useful for speciation studies advantages of SPE include easy handling of large sample volumes and convenience for automated sample processing.of metal–humic complexes in natural waters.14,15 In this work, Sephadex A-25 was used in combination with Chelex 100, in Groschner and Appriou1 employed Dowex 1X8, a strongly basic resin with quaternary amine functional groups, for the which the functional group is iminodiacetate. Although a great number of chelating sorbents have been used for the preconcen- preconcentration of anionic complexes in aqueous solutions, using 2 M HNO3 as eluent.They reported that the resin tration of trace metals from diVerent matrices,16,17 Chelex 100 has been a popular choice in many analytical applications.18,19 exhibited a high retention eYciency (98.8%) for ethylenediamine tetraacetate-bound copper, but for complexes with fulvic The performance of Sephadex A-25/Chelex 100 was compared with that of Dowex 1X8 Chelamine Metalfix for acid, 8-hydroxyquinoline, nitrilotriacetic acid and tryptophan, the recoveries of the copper species were rather poor.However, natural water samples. To evaluate the eYciency of the two- J. Environ. Monit., 1999, 1, 211–217 211column system for complete recoveries of metal ions, total water to obtain the required concentration.SPEX standard solutions were added to yield a concentration of 100 mg l-1 in concentrations were also determined by inductively coupled plasma mass spectrometry (ICPMS) or cold vapour atomic solution. BuVer solutions of 1 M ammonium acetate were added to adjust the pH. The 2 M HNO3 eluent was made by fluorescence spectrometry (CVAFS) for Hg.The experimental results were also compared with data calculated using equilib- diluting purified HNO3 (p.a., Merck, purified in the laboratory by subboiling distillation in a quartz still ) with Milli-Q water. rium speciation models. The resins used in the experiments were Chelamine Metalfix, (p.a., 40–80 mm, Fluka, Sigma-Aldrich Sweden AB, Experimental Stockholm, Sweden), Chelex 100 resin (analytical grade, 100–200 mesh, Bio-Rad Laboratories AB, Sundbyberg, Instrumentation Sweden), Dowex 1X8 (p.a., 100–200 mesh, Fluka) and DEAE Two diVerent ICPMS instruments (Table 1) were used for the Sephadex A-25 (Pharmacia Fine Chemicals AB, Uppsala, measurements, one equipped with a quadrupole mass filter Sweden).The columns used were made of poly(chlorotrifluor- (ICPQMS; VG Plasma Quad, VG Elemental, Winsford, UK) oethylene) (Malmo� Fluorocarbon, Malmo� , Sweden) and polyand the other with a high-resolution sector field mass spec- (etheretherketone) (SeQuant, Umea°, Sweden), and the dimentrometer (ICPSMS; ELEMENT, Finnigan MAT, Bremen, sions of the columns were 2 mm id×30 mm and 4 mm Germany).For the mercury determinations, a Millennium id×50 mm, respectively.The columns were equipped with Merlin System CVAFS instrument (PS Analytical, polypropylene frits to retain the resin, and were packed by Analys-konsult AB, Lidingo� , Sweden) was employed. using a slurry of ion exchange resin and Milli-Q water which In the preconcentration system, a peristaltic pump ( XV-4, was transferred with a pipette into the columst apply- Alitea, Sweden) was used.For the breakthrough tests, a ing suction. BischoV HPLC pump model 2250 (Coricon, Knivsta, Sweden) Prior to ICPMS determination of total analyte concendelivered sample to a BischoV Lambda 1010 UV detector for trations, all samples were acidified to obtain solutions in monitoring the eZuent absorbance at 254 nm.A Perkin-Elmer 0.14 MHNO3. Column eluates were diluted to obtain threefold Lambda 1 UV/VIS spectrophotometer (Perkin-Elmer, preconcentration in the solutions to be analysed by ICPMS. Norwalk, CT, USA) was used with 10 mm quartz cells for the For mercury determinations, KBr (p.a., Merck, Spa°nga, measurements of the absorbances at 400 nm to facilitate Sweden), KBrO3 (p.a., KEBO Lab AB, Spa°nga, Sweden) and estimation of the amounts of humic substances in the collected HCl (s.p., Merck) were added to the samples to concentrations water samples.of 0.47 mM, 1.43 mM and 1.5% (v/v), respectively. The samples were then allowed to stand for an hour. Immediately Reagents and solutions before analysis, 25 ml of 1.73 M hydroxylammonium chloride (p.a., KEBO Lab AB) was added to destroy residual oxidizing Standard solutions were prepared by diluting 1 mg ml-1 single agent and then a solution of 4% (w/v) SnCl2 (Merck) in 10% element standard solutions (SPEX Plasma Standards, Edison, (v/v) HCl was used to reduce Hg2+ to Hg0.The generated NJ, USA) with high purity water (Millipore, Milli-Q, Bedford, Hg0 was purged from the solution and detected by AFS.MA, USA). 1 M ammonium acetate buVer solutions (made in the laboratory by mixing 57 ml of anhydrous acetic acid with Preconcentration and elution 75 ml of purified 25% ammonia liquor and diluting to 1000 ml with high purity water; pH adjusted with HNO3 and NH4OH) All synthetic and natural water samples were handled on a were used to obtain the desired pH in the solutions.Model clean bench. Before preconcentration and elution, the anion solutions containing humic substances (Aldrich, Stockholm, exchange columns were flushed with 10 ml of 2 M HNO3, Sweden; batch number 8101816) were prepared from a stock followed by 20 ml of Milli-Q water. For the cation exchange solution containing 500 mg l-1 and diluted with high purity columns, 10 ml of 2 M HNO3 was passed, followed by 20 ml of Milli-Q water and, in the case of Chelex 100, 5 ml of 1 M Table 1 Operating parameters of the ICPMS instruments pH 6.0 ammonium acetate buVer.An aliquot of 20–30 ml of the filtered samples or model solutions was pumped through Parameter ICPQMS ICPSMSb the coupled columns with the aid of a peristaltic pump at a flow rate of 0.3 ml min-1 for the Dowex/Chelamine system Rf power/W 1348 1500 Sample uptake/ml min-1 0.8–0.9 0.1–0.3 and 1.0 ml min-1 for Sephadex/Chelex.Thereafter the columns Gas flow rates/l min-1 were disconnected from each other and the flow direction Coolant 14 15 reversed. The columns were washed with 5 ml of Milli-Q water Auxiliary 1.1–1.3 0.69 followed by elution with 2 M HNO3. Nebulizer 0.81–0.90 1.0–1.12 Ion lens settings a a Collection of water samples Torch Fassel torch Fassel torch Spray chamber Scott type, Scott type, Water samples were collected in 1998 from a bog-aVected double pass double pass drainage area near the Aitik copper mine in northern Sweden. Nebulizer Meinhard Meinhard Sample cone Nickel, Nickel, The samples were collected on the Julian dates 137 (17/05), 1.0 mm orifice 1.1 mm orifice 170 (19/06), 180 (29/06), 203 (22/07), 233 (21/08) and 260 Skimmer cone Nickel, Nickel, (17/09).In the field, the water samples were filtered through 0.7 mm orifice 0.8 mm orifice acid washed 0.45 mm filters (Millipore, type HA) and collected Acquisition mode Peak jumping E-scan into 1.5 l acid washed PTFE bottles. No preservation of the Dwell time/ms 10.24 10*, 20** samples was used to prevent any disturbance of the natural Samples per nuclide 3 30*, 25** Sweeps per replicate 20 15*, 15** equilibrium.Trip blanks were carried out to determine poten- Windows (%) tial contamination. In situ measurements of conductivity and Acquisition 30*, 100** pH values for the samples are given in Table 2. pH and Integration 30*, 80** conductivity measurements were performed using a Surveyor Search 30*, 60** II sonde (Hydrolab Corp., Austin, TX, USA).To obtain aAdjusted to obtain maximum signal intensity. bThe asterisks denote reliable pH values, the pH-electrode was allowed to stabilize the resolution mode: *low resolution and **medium resolution. for more than 30 min in the water. The temperature was also 212 J.Environ. Monit., 1999, 1, 211–217Table 2 Concentrations of humic substances (HS), pH, conductivities, temperatures and ionic strengths recalculated from conductivity20 of the natural water samples HS/ Conductivity/ Ionic strength/ Temperature/ Sample mg l-1 pH mS cm-1 10-4 °C 137 11.7 6.37 32.4 5.2 0 170 8.9 6.55 45.3 7.2 8.3 180 8.2 6.86 49 7.8 10.9 203 8.2 6.76 54.3 8.7 11.4 233 8.4 7.08 58 9.3 11.9 260 8.5 6.93 65.3 10.4 6.4 7967 14.1 6.52a b — b 7968 13.8 6.38a b — b 8891 12.8 5.81a b — b aMeasured in the laboratory. bNot measured.monitored as it aVects both the conductivity and pH. The pH and conductivity values in Table 2 are corrected for temperature dependence. Water samples (7967, 7968 and 8891) were also collected in the Lavsjo�n Lake, Sweden in September 1998.Estimation of humic substance concentrations in natural waters The concentrations of humic substances in the natural water samples were estimated using spectrophotometric measurement at 400 nm. Filtered (0.45 mm Millipore, type HA) standard solutions containing 0.5, 1.0, 2.0, 5.0, 10.0, 20.0 and 50.0 mg l-1 of the Aldrich humus were used for the calibration. Natural water samples were also filtered before the spectrophotometric determination. The results of the determinations of humic substances in the natural water samples are presented in Table 2.Equilibrium speciation modelling Fig. 1 Fractions of humic-bound metals (calculated as concentration retained on anion exchanger divided by the sum of the concentrations Individual models were established for each of the analytes, retained on the anion and cation exchangers) obtained using Sephadex calculations being performed using the free energy minimiz- (open bars) and Dowex (hatched bars).Errors bars are±one standard ation algorithm implemented in the computer program deviation (n=3), computed using the rules of error propagation for SOLGASWATER described by Eriksson.21 Thermodynamic including the uncertainties in both the determined humic-bound and summed concentrations.data were primarily taken from Gunneriusson and Sjo� berg,7 Lo�vgren and co-workers8,22 or the literature cited therein. In some cases, stability constants had to be adjusted to zero ionic Al and Co complexes. Three of the elements, Cr, Mo and V, strength conditions, which was accomplished using the are present, at least to some extent, as complex oxoanions in extended Debye–Hu� ckel expression proposed by Davies.23 natural waters, which means that the fractions retained on the Stability constants for metal–humic complexes were initially anion exchangers cannot be unambiguously classified as estimated from the formation constants of the corresponding organically bound. However, a significantly greater fraction oxalates, and the previously established relationship between of V is recovered by Sephadex than by Dowex (t=7.57; these values.7,8 As literature values from diverse sources may P>0.01).vary considerably, data were arbitrarily selected on the basis of their providing equilibrium speciation distributions that agreed, within a factor of two, with the experimental results.Humic substances were treated as a simple diprotic acid (H2L), and assuming one H2L group per 30 carbon atoms.7 The concentrations of humic substances, expressed in mg l-1, were estimated from the absorbance measurements described above and converted to mol l-1 active sites on the basis of the observation that Aldrich humic acid contains 50% by mass of carbon.24 Results and discussion Comparison of the ion exchange resins A compatween the two anion and the two cation exchange materials was performed.Selected results are depicted in Fig. 1. For Cd, Co, Mn, Ni and Sr, no significant diVerences in the amounts of humic complexes were obtained using either Sephadex or Dowex, so these data have been Fig. 2 Breakthrough curves for humic substances (HS) using omitted. With respect to precision, F-tests showed that Sephadex (&) and Dowex (+). Sample processed contained 5 mg l-1 HS. Sephadex was significantly better (P>0.05) than Dowex for J. Environ. Monit., 1999, 1, 211–217 213Table 3 Statistical comparison of the cation exchange materials in two-column speciation systems with respect to retained free metal ion concentrations and precisions (n=3) t-test F-test Cation exchanger P<0.01 P<0.02 P<0.05 P<0.01 P<0.02 P<0.05 Chelex Al, Ba, Sr, Zn Cd Fe Mo Ba Al, Cr, Cu Chelamine Ni — — — — — Table 4 Comparison of the speciation data with independent measurements of the total concentrations (n=3).Error terms are one standard deviation.Numbers in the Element column indicate the samples described in Table 2 Concentration/mg l-1 Elementa Sephadex Chelex Sumb Total t-statisticc Al (0.2) 137 32.58±0.29 22.02±0.33 54.60±0.44 68.57±0.79 -26.76*** 170 17.87±0.33 14.38±0.16 32.25±0.37 32.62±0.16 -1.59 180 10.81±0.37 7.75±0.16 18.56±0.40 18.92±0.20 -1.39 203 10.96±0.04 10.44±0.19 21.40±0.19 24.31±0.42 -10.93*** 233 5.53±0.22 6.48±0.17 12.01±0.28 12.18±0.06 -1.03 260 4.12±0.14 6.16±0.06 10.28±0.15 10.79±0.16 -4.03** (0.08) 7967 38.18±0.47 100.39±8.14 138.57±8.16 173.33±0.31 -7.37** (0.08) 7968 32.59±0.23 86.76±0.72 119.35±0.75 160.32±1.51 -42.14*** (0.08) 8891 31.11±5.05 93.81±6.70 124.92±8.39 168.51±0.73 -8.96** Ba (0.01) 7967 <0.003 5.04±0.30 5.04±0.30 4.81±0.01 1.33 7968 <0.003 5.04±0.05 5.04±0.05 4.62±0.04 11.72*** 8891 <0.003 4.75±0.17 4.75±0.17 4.78±0.08 -0.26 Cd (0.02) 137 0.02±0.01 0.02±0.01 0.04±0.01 0.04±0.03 0.00 170 0.02±0.01 0.04±0.02 0.06±0.02 0.04±0.01 1.55 203 0.03±0.01 0.03±0.01 0.06±0.01 0.05±0.01 1.22 233 0.03±0.01 0.10±0.01 0.13±0.01 0.13±0.03 0.00 Co (0.02) 170 0.22±0.02 0.29±0.02 0.51±0.03 0.05±0.01 25.20*** 180 0.07±0.02 0.03±0.01 0.10±0.02 0.04±0.01 4.65** 233 0.04±0.02 0.07±0.03 0.11±0.04 0.06±0.01 2.10 260 0.02±0.01 0.03±0.01 0.05±0.01 0.04±0.01 1.22 Cu (0.2) 137 5.63±0.15 0.31±0.02 5.94±0.15 5.81±0.19 0.93 170 3.13±0.07 0.30±0.02 3.43±0.07 3.56±0.21 -1.02 180 3.14±0.02 0.51±0.05 3.65±0.05 3.71±0.09 -1.01 203 2.10±0.06 1.44±0.01 3.54±0.01 4.11±0.06 -16.23*** 233 2.02±0.08 0.66±0.05 2.68±0.09 3.06±0.07 -5.77*** 260 2.08±0.04 1.48±0.05 3.56±0.06 4.02±0.21 -3.65 Fe (0.4) 7967 443.4±128.5 22.69±4.13 446.1±128.6 535.3±14.56 -1.19 7968 366.6±95.3 53.7±9.21 420.3±95.7 505.4±3.18 -1.55 8891 370.7±21.3 119.8±27.3 490.5±34.6 519.4±4.21 -1.44 Hgd (2.0) 7967 3.22±0.06 <0.67 3.22±0.06 3.60±0.04 -9.20*** 7968 2.96±0.06 0.77±0.00 3.74±0.06 3.30±0.06 9.09*** 8891 4.00±0.01 2.45±0.00 6.44±0.01 7.00±0.11 -9.12** Mn (0.3) 137 0.07±0.01 0.17±0.02 0.24±0.02 0.29±0.02 -3.06 170 0.44±0.02 0.48±0.02 0.92±0.03 0.21±0.01 38.89*** 233 0.05±0.02 0.52±0.04 0.57±0.05 0.54±0.02 0.96 (0.03) 7967 0.58±0.08 1.24±0.11 1.81±0.13 2.96±0.12 -11.22*** (0.03) 7968 0.43±0.09 0.95±0.11 1.38±0.14 2.92±0.06 -17.09*** (0.03) 8891 0.15±0.09 1.30±0.15 1.45±0.18 2.12±0.07 -6.07*** Pb (0.1) 203 0.23±0.01 1.03±0.03 1.26±0.03 1.12±0.05 4.16** Sr (2.0) 7967 <0.67 10.38±0.67 10.38±0.67 10.77±0.24 -0.94 7968 <0.67 10.91±0.29 10.91±0.29 10.70±0.09 1.18 8891 <0.67 10.18±0.75 10.18±0.75 10.66±0.34 -1.01 U (0.0005) 7967 0.115±0.009 0.005±0.001 0.120±0.009 0.139±0.002 -3.74 7968 0.098±0.001 0.004±0.000 0.102±0.001 0.134±0.001 -45.40*** 8891 0.113±0.007 0.006±0.002 0.119±0.007 0.136±0.001 -4.11 Zn (0.3) 137 0.46±0.38 4.01±0.07 4.47±0.39 4.55±0.58 -0.20 170 0.39±0.04 2.49±0.09 2.88±0.09 1.16±0.09 23.41*** 203 0.14±0.06 0.91±0.06 1.05±0.06 1.65±0.39 -2.63 (0.2) 7968 0.57±0.17 3.86±0.23 4.43±0.29 3.73±0.13 3.83 (0.2) 7968 0.21±0.03 6.15±0.97 6.36±0.97 3.46±0.24 5.03 (0.2) 8891 0.26±0.05 3.94±0.10 4.21±0.12 5.14±0.22 -6.56*** aDetection limits (mg l-1) in parentheses if not otherwise noted. bThe sum of the concentrations isolated by the Sephadex and Chelex columns.cNegative values indicate that the sum is less than the total concentration and positive values that the former is larger. The asterisks denote significant diVerences (two-tailed t-test) at the 95% (*), 98% (**) or 99% (***) confidence level. dMercury concentrations in ng l-1. 214 J. Environ. Monit., 1999, 1, 211–217Of the remaining seven elements, both anion exchange Sephadex resin, 360 ng humic complexes per ml resin are retained, whilst Dowex retains only 160 ng ml-1. materials provide statistically equivalent humic-bound fractions for Al, Fe and U. Clearly, U should be considered, along When comparing the cation exchange materials, Chelex shows a better performance than Chelamine, both with respect with Fe and Hg, as one of the elements exhibiting strongest association with humic substances in fresh water systems, to recovery and precision, for a range of elements (Table 3).The only exception is Ni, for which Chelamine exhibits a according to KerndorV and Schnitzer.10 Significantly greater fractions of the total Cu and Pb concentrations were recovered highly significantly (P<0.01) greater recovery.Both cation exchange columns proved to be equally viable for the other by the Sephadex column (t-test; P<0.01 for both elements). The Sephadex results are in accordance with the intermediate elements considered (Co, Mn, Pb, U, V). The Chelex column shows good recovery (84–100%) for many elements if the binding strengths observed for the humic complexes of these elements.Finally, for Ba and Zn, whose interactions with column is prewashed with 100 mM ammonium acetate pH 6.0 buVer before subsequent loading of the sample. Pai et al.18 naturally occurring organic ligands are expected to be negligible or weak, the use of Dowex resin results in serious observed that, when resins in the Na+, K+ or NH4+ form are used, it is advisable to apply external buVer systems, usually overestimations of the humic-bound fractions. For Ba, this can be explained by the low recovery of the free metal ions acetate.Chelamine shows good recovery for several elements, but has poor flow characteristics, flow rates as low as on the Chelamine column, as the sum of the concentrations collected by the anion and cation exchange materials was used 0.3 ml min-1 being needed to obtain a good recovery of trace metals.12 There is, however, no need for an external buVer to derive the humic-bound fraction. Chelamine is known to discriminate better against alkaline earth elements than system with Chelamine.As the anion exchange resins can be used at a flow rate of 1.0 ml min-1, it would only be time- Chelex.12 When measuring the absorbance of the acid Sephadex eluate consuming to use Chelamine instead of Chelex 100.With these results in mind, the conclusion can be drawn that Sephadex at 400 nm, a significant response was obtained. This finding suggests that the metal ions eluted are associated with humic and Chelex columns should be coupled if simultaneous preconcentration and separation of metal–humic complexes and free substances.When the same procedure was performed on the acid Dowex eluate, no (or very small compared to Sephadex) metal ions are to be accomplished. response was obtained, indicating eYcient retention of humic substances. An eVect visually observed is the discolouration Measurement of metal–humic complexes and free metal ions in of the resin, further supporting this assertion.There is a natural waters possibility that the humic substances are irreversibly retained on the resin, while the metal ions are eluted. When the waters were processed through the coupled columns, a fractionation of the total amount of metals in the water was The capacities of the anion exchange materials were investigated by performing a breakthrough test, using a model attained.Some of the fractions contained extremely low concentrations and are therefore reported as being less than the solution containing 5 mg l-1 Aldrich humic acid and 100 mg l-1 of the test elements. Breakthrough curves are shown in Fig. 2, limit of detection. Several results listed for the two-column method are below the detection limits for conventional where, in order to see small changes in the response, the wavelength used was 254 nm, as it provides higher sensitivity ICPQMS determinations, but quantitation is possible here due to the threefold preconcentration factor.Trip blanks were for humic substances than detection at 400 nm. For the Table 5 Thermodynamic data used in the calculations.Data are valid at 25 °C and zero ionic strength Metals/log bp, q, ra Species Al3+ Ba2+ Cd2+ Co2+ Cu2+ Hg2+ Mn2+ Pb2+ Sr2+ UO22+ Zn2+ MeOH+ -13.4 -10.08 -7.80 -3.40 -7.70 -13.2 -5.80 -9.00 Me(OH)2+ -5.00 Me(OH)2+ -10.52 Me(OH)3 -16.25 Me(OH)4- -23.46 Me(OH)2 -20.35 -15.00 -6.17 -17.10 -17.80 Me2(OH)22+ -10.60 -5.60 Me(OH)3- 28.30 Me2(OH)3+ -6.40 Me3(OH)42+ 23.90 -12.70 Me3(OH)5+ 15.60 MeH-1L -7.32 MeL+ -3.22 MeL2- -7.31 MeL 6.36 -4.26 -3.95 -3.10 2.73 -4.42 -3.84 -6.13 -2.07 -4.87 MeSO4 2.46 2.40 2.30 2.30 2.80 2.30 Me(SO4)22- 3.50 Me(SO4)34- 3.00 MeHCO3+ -2.90 Me(CO3)22- -18.36 Me(CO3)34- -29.04 MeCl+ 7.21 MeCl2 13.98 MeClBr 16.63 MeBr+ 9.54 MeBr2 17.91 aFormation constants are defined according to the general equilibrium: pH++qMen++rXm-uHpMeqXr p+nq-mr.J. Environ. Monit., 1999, 1, 211–217 215performed during the sampling procedure by processing To evaluate the capabilities of the two-column system for recovering trace elements, total concentrations determined Milli-Q water. These blanks were prepared and analysed as for the other samples and, for the elements studied, no utilizing accredited analytical methods were compared with the sums of values obtained from analyses of the eluates from contamination could be detected by ICPQMS. It can be seen in Table 4 that, for some analyte/matrix the Sephadex and Chelex columns.Quality control of total concentration determinations by ICPMS includes analysis of combinations, some fraction of the total concentration is not retained on either the Chelex or the Sephadex columns, which the riverine certified reference material SLRS-3 supplied by the Natural Research Council of Canada, for which accurate probably depends on losses during the washing step. It is, however, necessary to perform this washing step to ensure results are routinely obtained (results not shown here).The results of the statistical analysis are also reported in that the species eluted have been retained by the sorbent.To accomplish measurements of ultratrace levels of Hg and U in Table 4. The sum of the speciation results for Al is consistently lower than the total concentration. This also applies to the natural waters, a higher preconcentration factor would have to be used together with ICPQMS. Instead, other instrumental majority of data for Fe, Cu and Mn, although the diVerences are not always statistically significant.For the last two techniques (CVAFS and ICPSMS) were applied, thus providing the improved limits of detection required (2 ng l-1 Hg and elements, Groschner and Appriou1 found that 40–55% of Cu and 6–23% of Mn were present as complexes retained by C18 0.5 ng l-1 U, respectively, for direct measurement).Iron was measured in only three of the samples, as the technique used in their three-column speciation system. In the present twocolumn arrangement, at most 14% Cu and 53% Mn are not for six of the samples (ICPQMS) suVers from severe spectral interferences, e.g. 40Ar16O+ on 56Fe+. Significant portions of retained by either Sephadex or Chelex. It is improbable that this unretained fraction corresponds to neutral humic com- the total concentrations of most elements were retained on the Sephadex column (Table 2), illustrating the importance of plexes, because all water samples analysed had a low ionic strength and pH values above 5.8.Under such conditions, humic substances in determining the speciation of numerous trace metals in natural waters. humic substances are negatively charged.25 Thus, analyte escap- Fig. 3 Fractions of humic-bound metals, determined using the Sephadex anion exchange column (open bars) or calculated using equilibrium speciation models (hatched bars), in the water samples described in Table 2: (a) 137; (b) 170; (c) 180; (d) 203; (e) 233; (f ) 260; (g) 7967; (h) 7968; (i) 8891. Errors bars for the experimental measurements are±one standard deviation, computed using the rules of error propagation for including the uncertainties in both the determined humic-bound and total concentrations.The asterisks denote significant diVerences (twotailed t-test) at the 95% (*), 98% (**) or 99% (***) confidence level, and ‡ indicates that data were below the limit of detection for the experimental procedure. 216 J.Environ. Monit., 1999, 1, 211–217ing from the two-column system is more likely to be in the provides a reliable measure of the humic-bound fraction, and thus lends credibility to the results obtained. However, using form of neutral inorganic complexes, such as hydroxides, sulfates or carbonates.7 It was found to be impractical to only an anion exchange material for the separation of metal– humic complexes could give an incorrect estimation of the couple a third column in the present system, owing to the excessive back-pressure that compromised successful operation metals complexed by organic matter in the event that some neutral species exist in natural waters.To study neutral metal– of the peristaltic pump. As the concentrations of the humic substances were humus species, and if ICPMS is the detector of choice, a reversed phase material has to be used together with a mobile estimated, the pH determined and, in six of the samples, the ionic strength computed from the measured conductivities phase that will not cause interferences in the determination.(iv) Utilization of multi-element analytical techniques in using Langelier’s approximation20 (Table 2), calculations of the fractions of metal–humic complexes could be established studying trace element–humus interactions is also to be recommended. Even elements forming weak complexes with using the constants in Table 5.The sulfate concentrations in the samples were not measured; instead a reference value of humic substances will contribute to a depletion of available binding sites, potentially aVecting the toxicity and 0.1 mM7,26 for Swedish rivers was adopted.In Fig. 3, the measured and calculated fractions of the metal–humic com- bioavailability of other metals. plexes are compared. As the sums of the fractions of Al and Mn isolated by Chelex and Sephadex are significantly non- Acknowledgements quantitative, the separately determined total concentrations TFR (Technical Research Council ), Sweden is acknowledged were used to estimate the humic-bound fractions from the for financial support.SGAB Analytica, Lulea°, Sweden contrib- experimental results. Otherwise, the larger of the concenuted with both financial and technical assistance. Hans trations denoted sum and total in Table 4 was used to derive Hultberg, IVL, Go� teborg, Sweden is thanked for providing the experimentally determined fractions.The pH variation the Lavsjo�n samples used in this study. observed (5.81–7.08) in the samples analysed did not have any discernible eVect on the distribution of the metals. Instead, the changes in metal ion concentration and amount of humic References substances seem to be the most important factors.14 The 1 M.Groschner and P. Appriou, Anal. Chim. Acta, 1994, 297, 369. comparison of the experimental and calculated humic-bound 2 B. Aster, P. Burba and J. A. C. Broekaert, Fresenius’ J. Anal. fractions for Cd, Co, Mn, Pb and Zn in Fig. 3 reveals few Chem., 1996, 354, 722. significant diVerences, and it is interesting to note the impor- 3 M.Norde�n and E. Dabek-Zlotorzynska, J. Chromatogr. A, 1996, tance of humic substances for the speciation of Cd and Co, 739, 421. despite the weakness of these complexes. The equilibrium 4 K. G. Heumann, L. Rottmann and J. Vogl, J. Anal. At. Spectrom., speciation model is unreliable for Al and Hg, for which three 1994, 9, 1351. 5 L. Rottmann and K. G. Heumann, Anal.Chem., 1994, 66, 3709. and two possible humic complexes, respectively, have been 6 E. Tipping, Aquat. Geochem., 1998, 4, 3. proposed.7,8,22 For these elements, the aqueous speciation is 7 L. Gunneriusson and S. Sjo�berg, Nordic Hydrol., 1991, 22, 67. complicated by the formation of hydroxide and halide com- 8 L.Lo� vgren and S. Sjo�berg, Wat. Res., 1989, 23, 327. plexes, which suggests that greater attention should be paid 9 F.J. Stevenson, Humic Substances, Wiley, New York, 2nd edn., towards the input data requirements (i.e. determination of 1994, pp. 19–20. chloride and bromide concentrations) and to studying the 10 H. KerndorV and M. Schnitzer, Geochim. Cosmochim. Acta, 1980, 44, 1701. eVects of pretreatment of the Chelex column with buVer on 11 V. I.Esteves, N. M. A Cordeiro and A. Costa Duarte, Mar. the results. Chem., 1995, 51, 61. The experimentally determined fractions of humic-bound 12 S. Blain, P. Appriou and H. Handel, Anal. Chim. Acta, 1993, Cu were significantly lower than the modelled distribution in 272, 91. three of the six samples collected near the Aitik copper mine 13 B. R. Kim, V. L. Snoeyink and F. M. Saunders, J.Water Pollut. in northern Sweden. These samples constitute the latter half Control Fed. 1976, 48, 120. 14 C. Pettersson, B. Allard and H. Bore�n, Estuar. Coast. Shelf Sci., of a series in which the water conductivity increased with 1997, 44, 533. sampling date (Table 2). This would indicate that increasing 15 M. Hiraide, S. P. Tillekeratne, K. Otsuka and A. Mizuike, Anal. concentrations of major cations are competing for available Chim. Acta, 1985, 172, 215. binding sites on the humic substances.14 According to the 16 C. C. Huang and M. H. Yang, Anal. Chem., 1997, 69, 3930. model calculations, the 11 elements considered (Table 5) 17 K. Akatsuka, J. W. MacLaren, J. W. Lam and S. S. Berman, occupied as much as 80% of these sites. J. Anal. At. Spectrom., 1992, 7, 889. 18 S. Pai, P. Whung and R. Lai, Anal. Chim. Acta, 1988, 211, 257. 19 L. A. Ellis, J. Anal. At. Spectrom., 1998, 13, 631. Conclusions 20 V. L. Snoeyink and D. Jenkins, Water Chemistry, Wiley, New York, 1980. (i) For the quantitation of humic-bound metals in natural 21 G. Eriksson, Anal. Chim. Acta, 1979, 112, 375. waters, Sephadex is preferable to Dowex. The latter has a 22 L. Lo� vgren, T. Hedlund, L.-O. O� hman and S. Sjo�berg, Water lower capacity and irreversibly retains humic substances, and Res., 1987, 21, 1401. 23 C. W. Davies, Ion Association, Butterworths, London, 1962. incorrectly classifies Ba and Zn as forming fairly strong 24 S. Boggs, Jr., D. C. Livermore and M. G. Seitz, Rev. Macromol. complexes with naturally occurring organic ligands. Chem. Phys., 1985, C25, 599. (ii) Parallel determinations of total concentrations are 25 F. J. Stevenson, Humic Substances, Wiley, New York, 2nd edn., necessary, as in the preferred configuration of Sephadex/ 1994, p. 350. Chelex complete recoveries of Al, Cu, Mn and U were 26 K. B. Berner and R. A. Berner, The Global Water Cycle, Prentice not possible. Hall, Englewood CliVs, 1987, p. 397. (iii) The comparison of experimentally determined and calculated equilibrium distributions suggests that Sephadex Paper 9/01070J J. Environ. Monit., 1999,

ISSN:0960-7919    

DOI:10.1039/a901070j

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

Biomonitoring of contaminated mine tailings through age accumulation of trace metals in the bank vole (Clethrionomys glareolus) Adrian Milton and Michael Johnson* School of Biological Sciences, Life Sciences Building, University of Liverpool, Liverpool, UK L69 7ZB. E-mail: mjohnson@liv.ac.uk; Fax: +44 (0)151 794 4747; Tel:+44 (0)151 794 5291 Received 16th February 1999, Accepted 16th March 1999 Lead, zinc and cadmium were determined in a range of tissues from laboratory-bred bank voles (Clethrionomys glareolus) exposed to elevated levels of dietary zinc (124 mg g-1).The pelletised diet was derived from vegetation harvested from the surface of a revegetated tailings dam at a modern Zn–Pb mine. Exposure regimes to the contaminated diet were 16, 32, 64, 128 or 256 days. Elevated levels of dietary zinc were not reflected in the individual tissue or total body concentrations.Marginal age accumulation of lead and cadmium was evident in the liver (Pb) and kidney (Pb and Cd). Tissue residues did not attain toxicologically significant concentrations. Animals inhabiting the grassland are considered to be at low ecotoxicological risk with respect to trace metals. responses to metals.13 They do not, therefore, reflect the Aim of the investigation responses of a natural population.An experimental approach The use of small mammals as sentinels of environmental utilising wild-caught animals fed a diet prepared from contamipollution is a well-tested concept, with both wild and captive nated natural food items oVers the prospect of a better populations used historically.1–3 Analysis of tissues from live- representation of the natural dietary condition and also pretrapped animals from wild populations is commonly used to serves the genetic variability of natural populations as an indicate the extent of metal accumulation and also to ascertain experimental variable.tissue distribution. However, this approach does not take into The bank vole, Clethrionomys glareolus, is a small microtine account other unknown factors that aVect the accumulation rodent that is common throughout Western Europe.14 This of metals including disease, predation and, most importantly, species has been used to monitor environmental pollution age.4 Whilst the age of wild small mammals can be calculated, from a variety of technogenic sources.2,15,16 Moreover, there very approximately, by body weight,1 and estimated more have also been numerous laboratory-based investigations using accurately from teeth wear patterns and eye lens weight,5,6 this species, although these have tended to concentrate on controlled laboratory feeding trials enable a very accurate various aspects of population ecology.17 More recently, C.quantification to be made of the influence of age upon metal glareolus has been employed in laboratory studies of the dynamics. Furthermore, accurate monitoring of food con- accumulation and eVects of heavy metal exposure.8,18,19 sumption is also possible. This is strongly influential upon C. glareolus occupies a variety of habitats including lowmetal dynamics, not least because reduced food intake is an maintenance amenity grassland which is a common endpoint early indicator of excess dietary metals.7 of revegetation programmes on metalliferous mine tailings.20,21 A range of diVerent mammalian species has been used to It is also the only species of vole found in the Republic of investigate the accumulation and distribution of metals under Ireland, being first discovered in 1964. Although presently controlled laboratory conditions.However, many of these concentrated in the southwest of the country, the range of C. experiments have involved the administration of metals by glareolus is undergoing steady expansion22 at a rate of up to injection, by aqueous intake or through the formulation of 4.5 km yr-1.totally artificial diets with added mineral salts.8–10 These Ireland has a number of abandoned, active and proposed approaches provide an insight into the clinical eVects of metal metalliferous mines, some of which have already been encounions and their tissue distribution; however, they do not simulate tered by the expanding Irish population of C. glareolus.2 The the dietary conditions that prevail when animals encounter potential significance of metal residues in mine wastes to this metals via natural means as a result of environmental contami- relatively new Irish species needs to be assessed, especially as nation.In such cases, the speciation and related bioavailability there is a paucity of metal accumulation data for C. glareolus, of metal ions may influence their absorption from the alimen- notable exceptions being the bank vole populations resident tary tract, thus changing the accumulation and biological in contaminated Polish forests.23–25 This is in contrast to other eVects of the metal.11 Furthermore, environmental exposure common species of small mammals (e.g.Apodemus sylvaticus, to heavy metal contamination generally involves exposure to Microtus agrestis and Sorex araneus).more than one pollutant. This complicates the natural exposure To determine the potential toxicological eVects of a contamisituation by introducing antagonistic and synergistic inter- nated diet on captive small mammals, it is essential to establish actions between metals, e.g. zinc and copper.12 tissue distribution and the dynamics of metal accumulation Inbred strains of laboratory mice are often used in toxico- over time.This paper examines these issues using a captive logical experiments. These have a lower genetic variability breeding population of C. glareolus fed pelletised grass obtained from a revegetated surface at a modern metalliferous than field populations and may even show strain-specific J.Environ. Monit., 1999, 1, 219–225 219mine tailings facility in the Republic of Ireland. It is also the and certified reference materials (CRMs): BCR 185, bovine liver; BCR 186, pig kidney. Recoveries from both CRMs were aim of this paper to assess the long term viability of a consistently within the ranges: 95–110% for Zn, 90–108% for population of C. glareolus were it to eventually colonise the Pb and 91–107% for Cd.Where a suitable reference material grassland at the tailings facility in line with the ongoing was not available (hair, bone, muscle and carcass), spiked expansion of its territorial range in Ireland. samples were used to confirm method accuracy using acceptable recovery tolerances of ±10%. Description of experimental procedures All results were corrected to dry weight concentrations using predetermined conversion factors (carcass, 2.55; hair, 1.15; Bulk quantities (100 kg) of composite vegetation, mostly kidney, 3.40; liver, 3.58; muscle, 3.48).Total body concen- Festuca rubra and Agrostis capillaris, were collected from the trations (TBCs) were calculated by pooling individual tissue revegetated surface of a tailings dam attached to a modern results with carcass values.Wet weights were used for compari- Zn–Pb mine, and also from an uncontaminated control site.sons of tissues and body weights since these are diagnostic of The material was then pelletised and, following pilot feeding renal or hepatic oedema.30 trials, upgraded with supplementary animal fats and carbo- Arithmetic means and the standard error of the mean hydrates.This was achieved without appreciably reducing the (SEM) were calculated using Microsoft EXCEL. To determine dietary concentration of metals or changing its fundamental diVerences between sexes within treatments, t-tests (unequal nature relative to the composition of the diet available to wild variance assumed: p<0.05) were used.There were very few animals that might invade the tailings surface. The final pellet significant diVerences in the concentrations of Cd, Pb and Zn formulation comprised 80% grass, 16% animal fat and carbobetween the sexes on either diet or exposure period, and so all hydrate and 4% vitamins and minerals. data for the two sexes in all treatments were pooled for further Concentrations of Zn (contaminated: 123.8±4.0 mg g-1 dry statistical analysis.Datasets were subjected to two-way Analysis wt; control: 66.3±3.2 mg g-1 dry wt), Pb (3.38±0.2 mg g-1 of Variance (ANOVA), using diet type and exposure period dry wt and 3.69±0.2 mg g-1 dry wt) and Cd (0.34±0.1 mg g-1 as factors. Where significant diVerences (p<0.05) were dry wt and 0.25±<0.01 mg g-1 dry wt) in the feed pellets detected, least significant diVerences (LSDs) were calculated.31 were determined by flame atomic absorption spectrometry (AAS).The concentration of zinc diVered significantly Results ( p<0.001) between the contaminated and control diets, whilst the concentrations of Pb and Cd were very similar for the two Total body wet weights are given in Table 1.Increases in body diets. This is despite the substantially diVerent soil concen- weight in line with the exposure period are to be expected, as trations of 6000 mg kg-1 Zn, 2000 mg kg-1 Pb and 22 mg kg-1 is the sexual dichotomy of animal weights in line with normal Cd at the tailings dam as compared to 160 mg kg-1 Zn, mammalian growth patterns. Comparisons can therefore only 40 mg kg-1 Pb and <1 mg kg-1 Cd at the control site 26.be made between diets at each exposure juncture for each sex. Mature C. glareolus were obtained from a long-established Only males showed significant diVerences in total body weight outbred colony housed at the University of Liverpool, and between control and contaminated diets, with 64 day prospective breeding pairs were matched by age.Breeding ( p<0.001) and 128 day ( p<0.01) animals fed the contamistock animals were fed standard rat/mouse diet (Special Diets nated diet heavier than the controls. However, males exposed Service, Cambridge, UK) and given tap water ad libitum. for 256 days showed the reverse trend, with control animals A total of 140 (70 male, 70 female) oVspring were separated heavier than those on the contaminated diet (p<0.05).The from parent animals when weaned at 18 days and transferred weights of all animals were, however, similar to those given into individual cages supplied with de-ionised water and the by Baker and Clarke17 for laboratory-bred C. glareolus of standard rodent diet. After 7 days, individuals were randomly comparable ages. assigned to either the contaminated or control diet, 7 males Expressing liver and kidney weights as a percentage of total and 7 females to each of 5 experimental exposure periods.body weight (relative organ weight) gives a good indication De-ionised water was provided ad libitum. Food consumption of metal toxicosis,30 and these data are shown in Table 1. was monitored daily throughout, and animals were weighed Comparing all data within each exposure period and between daily for the first 16 days after which weighing was undertaken diets revealed no significant diVerences ( p>0.05); however, three times a week.All animals were kept under standard there were exposure-based diVerences for both the liver and laboratory conditions (20±1 °C, 12 h light–dark cycle).kidney (p<0.001), and the LSDs are given in Table 1. Exposure periods of 16, 32, 64, 128 and 256 days were Zinc concentrations in the tissues of C. glareolus are prechosen to allow accurate determination of age accumulation, sented in Tables 2–6 together with LSDs ( p<0.05 and p<0.01) with the longest treatment period being between the average for sample means. The hierarchy of concentrations was similar and maximum life span for the species in the field.14,27 At the for both diet regimes and all exposure periods: end of their exposure, animals were killed humanely and femur>hair>liver>kidney>muscle.A number of statistifrozen until dissection in order to prevent tissue lysis and cally significant diVerences were recorded between exposure metal redistribution.period and diet types for femur, hair, kidney and muscle Animals were dissected using stainless steel instruments, tissues, the exception being liver (Table 5). with weighed samples of hair, liver, kidney, muscle, bone Bone concentrations of zinc (Table 2) were characterised by (femur) and the residual carcass ( less gut contents) being cold significant diVerences based on the exposure period (p<0.05). digested overnight prior to hot digestion with concentrated However, only animals receiving the control diet, for the two nitric acid (MERCKA, Poole, Dorset, UK, Aristar grade) at longest exposures (128 and 256 days), were significantly higher 120 °C for 1 h.Prior to digestion, femurs were cleaned enzy- than all other exposure/diet combinations.Importantly, femur matically using papain solution to remove all soft tissue.28 concentrations of zinc for animals receiving the contaminated Acid-washed glassware was used at all times. Zn concentrations diet rarely exceeded those of animals maintained on the control were determined using conventional flame AAS, whilst Pb and diet. Concentrations of zinc in hair (Table 3) diVered signifi- Cd were measured by flameless AAS using a graphite furnace cantly (p<0.001), but therewas no clear pattern of accumulation fitted with Zeeman background correction.29 Full details of based on either exposure period or diet type.Similarly, kidney the methods and furnace programming are described in residues of zinc (Table 4) showed occasional significant diVer- Milton.26 ences (p<0.001) according to exposure period and diet type, but with no discernible pattern of exposure or dietary influence. Sample batches were analysed with double reagent blanks 220 J.Environ. Monit., 1999, 1, 219–225Table 1 Total body weight and relative liver and kidney weights Exposure/days Diet Sex Total bodyb Liver (%)a Kidney (%)a 16 Control M 15.8±0.61 5.7±0.22 1.7±0.05 F 15.3±0.48 5.8±0.34 1.8±0.05 Contaminated M 16.3±0.58 5.5±0.10 1.6±0.04 F 14.9±0.26 5.7±0.37 1.6±0.04 32 Control M 16.2±0.38 5.4±0.12 1.6±0.04 F 16.8±0.38 5.7±0.16 1.6±0.08 Contaminated M 16.1±0.62 4.8±0.18 1.5±0.05 F 15.8±0.71 5.1±0.07 1.6±0.11 64 Control M 16.2±0.33 5.5±0.15 1.5±0.03 F 16.3±0.22 5.2±0.16 1.4±0.04 Contaminated M 20.2±0.73 4.8±0.22 1.5±0.07 F 16.1±0.78 6.2±0.60 1.7±0.10 128 Control M 17.3±0.25 4.1±0.15 1.4±0.08 F 15.5±0.37 4.6±0.20 1.6±0.12 Contaminated M 20.1±0.40 4.5±0.31 1.5±0.06 F 17.1±0.66 4.6±0.21 1.5±0.05 256 Control M 23.8±1.29 5.1±0.22 1.4±0.03 F 17.8±0.60 5.3±0.27 1.5±0.04 Contaminated M 21.6±0.58 4.8±0.11 1.4±0.04 F 17.8±0.79 5.1±0.22 1.5±0.04 LSD ( p<0.05)c 0.67 0.18 LSD ( p<0.01) 0.88 0.24 aExpressed as a percentage of total body weight.bData as g wet wt: n=7. cLeast significant diVerence at (stated level of probability). Table 2 Concentrations of zinc, cadmium and lead in bone (femur) of Clethrionomys glareolusa Diet Exposure/days Zinc Cadmium Lead Control 16 220.0±5.6 0.68±0.03 6.19±0.38 32 226.4±9.3 0.16±0.01 1.99±0.36 64 230.2±4.0 0.28±0.05 2.48±0.40 128 257.2±4.1 0.35±0.01 3.63±0.26 256 261.1±7.4 0.15±0.04 0.99±0.14 Contaminated 16 222.9±28.1 0.55±0.07 5.44±0.56 32 248.0±6.6 0.10±0.01 2.12±0.29 64 222.1±3.8 0.29±0.04 2.77±0.39 128 246.9±9.9 0.31±0.02 4.08±0.25 256 241.2±6.7 0.12±0.01 2.70±0.12 LSD (p<0.05)b 30.6 0.10 0.94 LSD (p<0.01) 40.4 0.13 1.24 aAll data in mg g-1 dry wt±SEM; n=14.bLeast significant diVerence at (stated level of probability).Table 3 Concentrations of zinc, cadmium and lead in hair of Clethrionomys glareolusa Diet Exposure/days Zinc Cadmium Lead Control 16 228.5±3.1 0.05±0.01 0.20±0.03 32 209.0±7.0 0.03±0.0 0.23±0.04 64 207.0±13.3 0.02±0.0 0.31±0.04 128 233.9±6.7 0.05±0.01 0.39±0.06 256 230.1±4.5 0.05±0.0 0.34±0.04 Contaminated 16 206.4±4.2 0.05±0.01 0.42±0.07 32 203.3±11 0.04±0.0 0.26±0.04 64 205.6±3.7 0.03±0.0 0.38±0.08 128 224.7±6.1 0.05±0.01 0.36±0.04 256 199.5±2.0 0.03±0.0 0.30±0.03 LSD (p<0.05)b 19.7 0.02 0.14 LSD (p<0.01) 26.0 0.02 0.19 aAll data in mg g-1 dry wt±SEM; n=14.bLeast significant diVerence (at stated level of probability). Control animals did, rather surprisingly, tend to have higher Overall, the concentrations of zinc were similar to those described for C.glareolus collected from a number of free- overall zinc levels than those fed the contaminated diet, but the diVerences were only significant for the 64 and 256 day living populations (Table 7), with only kidney residues being notably lower in animals from the present study. The data exposures. Muscle zinc concentrations (Table 6) diVered between diets only with respect to animals on the 128 day were also similar to those reported for other ecologically related species (e.g.Microtus agrestis).15 treatment where concentrations were significantly higher than for all other exposure/diet pairings ( p<0.05). Lead concentrations in experimental C. glareolus are J. Environ. Monit., 1999, 1, 219–225 221Table 4 Concentrations of zinc, cadmium and lead in kidneys of Clethrionomys glareolusa Diet Exposure/days Zinc Cadmium Lead Control 16 85.4±1.2 0.09±0.01 0.30±0.03 32 100.3±2.3 0.14±0.01 0.33±0.03 64 97.9±1.9 0.23±0.01 0.50±0.04 128 95.7±4.2 0.24±0.02 0.43±0.08 256 100.2±1.6 0.74±0.05 0.36±0.04 Contaminated 16 85.3±4.3 0.14±0.03 0.36±0.03 32 99.2±4.9 0.19±0.01 0.30±0.05 64 84.9±1.4 0.22±0.01 0.60±0.06 128 90.2±3.0 0.28±0.03 0.38±0.02 256 85.6±1.6 0.78±0.06 0.42±0.04 LSD (p<0.05)b 8.2 0.08 0.13 LSD (p<0.01) 10.9 0.11 0.17 aAll data in mg g-1 dry wt±SEM; n=14.bLeast significant diVerence (at stated level of probability). Table 5 Concentrations of zinc, cadmium and lead in liver of Clethrionomys glareolusa Diet Exposure/days Zinc Cadmium Lead Control 16 120.2±2.7 0.06±0.01 0.17±0.01 32 118.9±2.4 0.06±0.01 0.28±0.03 64 121.1±8.5 0.15±0.01 0.49±0.06 128 126.8±7.0 0.12±0.00 0.62±0.12 256 115.4±3.7 0.25±0.01 0.87±0.06 Contaminated 16 120.4±5.2 0.05±0.00 0.30±0.02 32 129.9±2.2 0.08±0.00 0.39±0.04 64 125.1±2.3 0.17±0.01 0.50±0.05 128 120.0±3.8 0.15±0.01 0.53±0.06 256 111.0±3.2 0.23±0.01 0.97±0.11 LSD (p<0.05)b 12.8 0.02 0.18 LSD (p<0.01) 16.9 0.03 0.24 aAll data in mg g-1 dry wt±SEM; n=14.bLeast significant diVerence (at stated level of probability). Table 6 Concentrations of zinc, cadmium and lead in muscle of Clethrionomys glareolusa Diet Exposure/days Zinc Cadmium Lead Control 16 44.6±1.7 0.03±0.01 0.16±0.02 32 49.5±3.2 0.03±0.00 0.16±0.03 64 44.9±1.9 0.02±0.00 0.17±0.03 128 55.5±4.6 0.03±0.00 0.14±0.01 256 48.4±4.1 0.03±0.00 0.09±0.01 Contaminated 16 47.3±1.1 0.03±0.01 0.11±0.02 32 47.2±1.8 0.03±0.00 0.19±0.07 64 40.6±1.8 0.02±0.00 0.12±0.02 128 53.8±3.4 0.04±0.01 0.11±0.01 256 42.1±4.1 0.03±0.00 0.13±0.02 LSD (p<0.05)b 8.5 0.01 0.08 LSD (p<0.01) 11.2 0.01 0.10 aAll data in mg g-1 dry wt±SEM; n=14.bLeast significant diVerence (at stated level of probability). presented in Tables 2–6. The hierarchy of concentrations was ated and diVerences between subsequent exposures were insignificant or only marginally significant ( p<>0.05).Significant broadly similar for the control and contaminated diets. The overall pattern of tissue concentrations for both diets and all diVerences between diVerent exposure periods (p<0.001) were detected for hair and kidney concentrations (Tables 3 and 4), exposures was: femur>kidney, liver>hair>muscle.Bone (femur) contained the highest concentration of lead (Table 2), although the fluctuating pattern of accumulation indicated no clear directional influence of either diet or age. highlighting the importance of the skeleton in the metabolic management and depositional storage of lead even at very low There were significant exposure based diVerences in the concentration of lead in the liver ( p<0.001), with a progressive dietary levels.Exposure-based statistically significant diVerences were recorded for all the tissues analysed (p<0.001), increase over time for both control and contaminated diets (Table 5). This notable age accumulation of lead did not, with the exception of muscle (Table 6).There were significant diVerences ( p<0.05) in the concen- however, reveal any divergence between the two diets either in the rate or magnitude of lead accumulation for equivalent trations of lead in the femur between exposure periods for the same diet (Table 2). The main diVerence was between the 16 exposure periods (p>0.05). Concentrations of lead in C.glareolus from a wide range of day and each of the 32 to 256 day exposure periods, the former being significantly higher in all cases (p<0.05). After field and laboratory investigations are given in Table 7. Values derived from the present study are mostly lower than those this initial early peak value at 16 days, concentrations fluctu- 222 J. Environ. Monit., 1999, 1, 219–225Table 7 Mean zinc, lead and cadmium concentrations (mg g-1 dry wt) in the tissues of Clethrionomys glareolus from laboratory (L) and freeliving (F) populations Metal, exposure and site Pop.Kidney Liver Bone Hair Carcass Ref. Zinc Mixed forest—autumn F 73.0 23 Deciduous forest—autumn F 73.0 23 Mixed control forest F 102.8 23 Coniferous forest N F 145 118 24 Coniferous forest D F 156 120 243 148 24 Control forest F 143 115.8 190 169 153 24 4 km from smelter F 74.1 16 90 km from smelter F 68.0 16 Mine site F 101 85.0 282 32 Control grassland F 158 133 200 32 Lead Mixed forest—autumn F 5.7 23 Deciduous forest—autumn F 5.4 23 Mixed control forest F Not detected 23 Coniferous forest N F 14.3 8.8 12.7 6.1 9.7 24 Coniferous forest D F 13.1 3.5 2.9 20.3 24 Control forest F 14.0 3.9 12.3 2.3 2.5 24 Near smelter F 2.3 16 Control grassland F 1.5 16 Mine site F 46.5 10.4 311 32 Control grassland F 8.5 7.0 10.3 32 20 days fed 80 mg g-1 grain L 35.7 8.8 17.1 1.2 166 18 20 days fed 300 mg g-1 grain L 17.3 14.4 14.2 2.0 209 18 Control grain L <0.1 <0.1 <0.1 0.6 1.1 18 Control grassland F 12.0 33 Shooting range F 15.8 5.1 26.3 30 Control grassland F 5.9 2.8 1.9 30 Control grassland F 1.1 0.6 0.1 2 0.8 km from road F 5.0 7.5 34 Cadmium Mixed forest—autumn F 9.2 23 Deciduous forest—autumn F 7.3 23 Coniferous forest N F 2.4 1.4 0.9 0.2 0.1 24 Coniferous forest D F 26.3 4.2 0.8 24 Control forest F 3.0 0.6 0.5 0.2 0.3 24 Near smelter F 21.8 16 Control site F 0.6 16 Smelter waste F 16.8 5.1 32 Control grassland F 4.6 1.1 32 20 days fed 10 mg g-1 grain L 25.5 8.8 7.7 2.5 10.6 18 20 days fed 40 mg g-1 grain L 64.5 17.8 4.9 2.1 28.7 18 Control grain L 4.2 0.3 3.6 1.3 2.0 18 Low Cd pellets (ND) L Not detected 19 High Cd pellets (8.6 mg Cd g-1) L 13.9 19 reported previously, the exception being data for the femur animals from the 16 day exposure period having the highest concentration and those from the 32 to 256 day exposures where the range in treatment means for the present dataset (Table 2) is such that some exposure/diet combinations are having lower values, although not consistently related to the duration of exposure. above those reported elsewhere for control animals.However, no femur lead concentrations from the present study even Concentrations of cadmium in hair (Table 3) showed a significant diVerence between the 64 day and other exposure approach the much higher values reported for animals fed contaminated diets or inhabiting contaminated field sites else- regimes (p<0.001).Similar statistically significant (p<0.01), but biologically insignificant, diVerences were detected for where (Table 7). Cadmium concentrations in the tissues of C. glareolus are cadmium levels in muscle (Table 6).The concentrations of cadmium reported from previous summarised in Tables 2–6. The hierarchy of concentrations for both diets for the majority of exposure periods was: femur, studies of C. glareolus are generally higher than those presented here (Table 7). Most importantly, cadmium concentrations in kidney>liver>hair, muscle.There is, however, evidence of age accumulation of cadmium in the kidneys and liver, such the kidney, which is known to have a distinctive role in the storage of cadmium,35 were significantly lower than all the that for the longer exposure periods renal cadmium exceeded that of the femur (Tables 4 and 5). Importantly, with the values reported previously. Total body concentrations of zinc, lead and cadmium reflect exception of the femur (p<0.05), there were no significant diVerences in the cadmium concentrations of any tissue when the patterns found for the individual tissues with no overall significant diVerences between the two types of diet ( p>0.05); compared between diets within any exposure period ( p>0.05).The significant diVerence in femur residues is based almost however, zinc and lead showed exposure-related diVerences ( p<0.01). For zinc, the diVerences appear random, with no entirely on the high cadmium levels in the 16 day exposure animals on both diets (Table 2).Femur concentrations of trend apparent based on the duration of exposure. In contrast, total body lead concentrations showed evidence of age cadmium followed a similar pattern to that of lead, with J.Environ. Monit., 1999, 1, 219–225 223accumulation that is probably strongly influenced by, and None of the experimental animals accumulated metals to significant levels in the tissues selected for analysis. Of particu- parallels, the pattern seen in the liver and kidney. However, the generally low concentrations in the individual tissues are lar importance is the consistency of zinc concentrations, which aYrms the presence of mechanisms that maintain the optimum reflected in the total body concentrations of zinc, lead and cadmium which are lower than those reported elsewhere.15,25 environment for the metabolic activity of cells, despite a doubling of the dietary intake burden.26 The MTs, cysteinerich, low molecular weight proteins that bind zinc as well as Discussion cadmium and other metals,41–43 are important in the maintenance of homeostatic zinc concentrations.Dietary ingestion is recognised as the primary source of environmental heavy metal exposure in small mammals.1,36 Generally, the observed concentrations of all three metals were below those reported for animals from a range of Carnivorous small mammals (e.g. shrews) are regarded as being most ‘at risk’ from accumulated heavy metals since their laboratory and field investigations (Table 7).Significantly, the renal concentrations of lead did not exceed the 25 mg g-1 dry invertebrate-based diet can contain high levels of metals.35 However, other animals (e.g. voles), which form an important wt considered to be a critical concentration for subclinical toxicity in wild mammals.30 Similarly, all concentrations of part of grassland ecosystems, can also attain high tissue concentrations.15 Such rodents are important prey items for cadmium were below the lowest observable adverse eVect levels (LOAELs) for residues in the kidneys of adult small birds of prey and larger mammals,14 and provide a vehicle for trophic level transport and for food-chain transfer of metals.mammals which range from 350 to 1000 mg g-1 dry wt.31 A lower threshold concentration of 30 mg g-1 wet wt (110 mg g-1 This has implications for the community structure of small mammal populations as ecotoxic metals can instigate changes dry wt) has been reported as inducing subclinical renal histopathological changes,36,44 but, again, this is over 100 times higher in species abundance.Localised extinction is even possible, through diminished habitat quality that causes reductions in than the peak cadmium concentration from the present study. More pertinent, however, is the fact that free-living C. glareolus fecundity or longevity, as well as through direct toxic eVects.For example, Kataev et al.37 found that C. glareolus was inhabiting areas impacted by smelter emissions exhibit increased levels of proteinurea linked to renal-tubular dysfunc- absent from areas severely impacted by emissions from a copper–nickel smelter due to reductions in food abundance tion at kidney cadmium concentrations as low as 4 mg g-1 wet wt (14 mg g-1 dry wt).16 Should this recent lowering of and possibly direct toxic eVects.A range of nephrotoxic eVects indicative of renal-tubular damage have also been found in C. LOAELs continue, then the threshold at which clinical eVects of cadmium may be recorded will rapidly converge with glareolus inhabiting a cadmium-contaminated area.16 Heavy metals provoke a wide range of toxic responses in background concentrations of the metal in mammalian tissues.The very low levels of lead and cadmium in the soft tissues mammals including reductions in growth, immunosuppression and fecundity, as well as the induction of carcinogenesis, of animals in this study surely reflect the low initial levels in the dietary pellets derived from the indigenous vegetation.embryotoxicity and teratogenesis.33 Many factors are capable of modifying the reaction of individuals to dietary insult, most Also of considerable importance to the low body burdens is the elimination under experimental conditions of the passive notably age and degree of acclimation, the chemical form of the metal (e.g. mercury and methylmercury) and interactions ingestion of soil as an exposure route.45 Soil ingestion by small omnivorous mammals has been estimated at 250– with other dietary compounds.12 The pelletised diets formulated for this feeding trial were 500 mg day-1.45–47 This route of contaminant intake is exacerbated under field conditions by the tunnelling habit of C.structured around the same grass species, Festuca rubra, harvested from clean and contaminated sites.The subsequent glareolus which, coupled with grooming, increases the risk of ingestion of soil particulates. Furthermore, under field con- addition of carbohydrates and fats to improve the nutritional quality of the diet was standardised for both diets, resulting ditions, animals stressed by the ingestion of toxic materials resort to the ingestion of soil as a counter-measure to reduce in final pellets that diVered only in their zinc loading and not lead or cadmium.Moreover, this adjustment generated a diet the absorption of toxins (e.g. tannins) from the digestive tract.48 In circumstances in which the soil itself is the vehicle with carbohydrate, protein and energy qualities more closely matched with field conditions, wherein invertebrates, fruits of pollutant exposure, such ingestion exacerbates the problem triggered by dietary metals.Also eliminated from the diet of and seed endosperm are minority food items.2,26 Environmental and dietary-related stress induce changes in laboratory C. glareolus are a range of vegetation types, including leaf litter, fungi and invertebrates but these represent a the metabolic rates of animals.39,40 The elevated zinc level in the contaminated diet has stress potential because increases in small proportion of the diet22 and make an insignificant contribution to the dietary metal flux.food intake and metabolic rate may be required to meet the demand for maintenance of homeostatic levels of zinc, possibly The exposure-based changes in the lead concentration of the femur are attributed to the eVects of development- by the induction of metallothionein (MT) synthesis.This could explain the marginally increased consumption of con- dependent factors. The high initial concentrations in the 16 day treatment may be the result of the pre-parturition transfer taminated diet by animals on the longer exposure regimes (64, 128 and 256 days).26 Animals on the shorter trials (16 and 32 of metals from pregnant females to the developing foetuses, coupled with a post-parturition transfer via milk.49 Pregnant days) were probably already consuming food at a maximum rate to obtain the energy and raw materials necessary for females were maintained on a standard diet with an extremely low lead concentration (1.1 mg g-1; Special Diets Service) normal growth.Animals on the contaminated diet were expected to be most which eVectively excludes the food supply as an exposure route. However, their drinking water contained suYcient dis- at risk of exhibiting renal and hepatic oedema both of which are early indicators of clinical plumbism.30 However, the high solved lead (50 mg l-1) to result in marginal accumulation in the parents and their oVspring.Lead is often found in high metabolic rate and especially the low dietary concentration of lead, and corresponding tissue values, yielded relative organ concentrations in the skeletal tissue of mammals, and this is ascribed to the chemical interaction between calcium and lead weights for animals which diVered between diets only for the liver of 32 day exposed animals.The prognosis is against renal which have common transport systems that lead to the incorporation of lead into the bone matrix during ossification.49 or hepatic dysfunction since relative organ weights for both dietary regimes were similar to those reported for animals Ossification, and the passive deposition of lead, is maximal during the peak growth phases including weaning. Suckling inhabiting a clean site.30 224 J.Environ. Monit., 1999, 1, 219–2252: Health EVects, ed. J. O. Nriagu, Wiley, New York, 1980, females also re-mobilise bone material during lactation to pp. 61–70. ensure an adequate supply of calcium to their oVspring. This 8 R. Swiergosz, M. Zakrzewska, K. Sawicka-Kapusta, K. Bacia and may inadvertently increase the dietary exposure of the pre- I.Janowska, Ecotox. Environ. Safety, 1998, 41(2), 130. weaned pups to bioavailable lead.49 9 K. Arizono, J. Sakamoto, M. Mikajiri, A. Murashima and T. Weaned animals were given a lead-free water supply causing Ariyoshi, Trace Elem. Med., 1993, 10, 80. 10 J. A. Cooke, S. M. Andrews and M. S. Johnson, Water, Air, Soil the grass pellets to be the sole lead exposure route.The Pollut., 1990, 51, 55. pelletised grass had a higher lead content than the standard 11 A. Davis, M. V. Ruby and P. D. Bergstrom, Environ. Sci. Technol., diet, but the strongly protein-bound nature of lead in the 1992, 26, 461. pelletised vegetation will have mitigated against absorption, 12 C. T. Walsh, H. H. Sandstead, A. S. Prasad, P.M. Newberne and thus reducing the overall retained burden of lead as body P. J. Faker, Environ. Health Perspect., 1994, 102, 5. weight increased in line with exposure. It is also possible that 13 C. Farr and D. M. Hunt, Biochem. Gen., 1989, 27, 199. 14 G. B. Corbet and H. N. Southern, The Handbook of British the downward trend in femur lead levels over time is linked Mammals, Blackwell Scientific, Oxford, 2nd edn., 1977.to the normal mechanisms of bone resorption and mineral 15 S. S. Talmage and B. T. Walton, Rev. Environ. Contam. Toxicol., re-mobilisation,50 whereby bone lead is replaced by calcium 1991, 119, 47. and relocated or even eliminated from the body. 16 P. E. LeZer and N. E. I. Nyholm, Ambio, 1996, 25, 417. There is compelling evidence for the age accumulation of 17 J.P. Baker and J. R. Clarke, in The UFAW Handbook on the Care cadmium and lead in the liver and kidneys of C. glareolus. A and Management of Laboratory Animals, ed. T. B. Poole, Longman, Bath, 1987, pp. 331–345. positive correlation exists in both tissues between exposure 18 K. Sawicka-Kapusta, A. Go� recki, R. Swiergosz, G. Juszczak, M. period and cadmium concentration [r=0.82 ( liver) and r= Mielczarek and B.Wo� jcik, Ekologia Polska, 1987, 35, 399. 0.88 (kidney); p<0.001; n=140], whilst a similar correlation 19 R. F. Shore, D. G. Myhill, E. J. Routledge and A. Wilby, Arch. exists for lead in the liver (r=0.69; p<0.001; n=140). Environ. Contam. Toxicol., 1995, 29, 180. Although this demonstrates the propensity of lead and cad- 20 M.S. Johnson, J. A. Cooke and J. K. W. Stevenson, in Mining mium in these target organs to accumulate over time, the and its Environmental Impact, ed. R. E. Hester and R. R. Harrison, Royal Society of Chemistry, Cambridge, 1994, pp. 31–48. tissue concentrations are dramatically lower than those 21 S. J. Burkinshaw, M. S. Johnson, J. V. Bramley and reported to cause a toxicological response. Nevertheless, the P.J. Huxtable, in Minerals, Metals and the Environment II, pattern of age accumulation confirms the importance of these Institute of Mining and Metallurgy, London, 1996, pp. 453–461. tissues as repositories for lead and cadmium and mirrors the 22 C. M. Smal and J. S. Fairley, Mamm. Rev., 1984, 14, 71. pattern of accumulation observed in wild populations of C. 23 T. Wlostowski, Ekologia Polska, 1987, 35, 115. glareolus and other small mammals, albeit at significantly 24 K. Sawicka-Kapusta, R. Swiergosz and M. Zakrewska, Environ. Pollut., 1990, 67, 315. lower concentrations.4,16 25 M. Zakrzewska, K. Sawicka-Kapusta, A. Perdenia and A. Wosik, The housing of animals in laboratory conditions, whilst Sci. Tot. Environ. (Suppl.), 1993, 133, 167.allowing the accurate monitoring of growth, food intake and 26 A. M. Milton, PhD Thesis, University of Liverpool, 1997. metal dynamics, does introduce diVerences to the contaminant 27 D. G. L. Innes and J. S. Millar, Mamm. Rev, 1994, 24, 179. exposure dynamics of the external environment. Most import- 28 E. Pankakoski and I. Hanski, Ann. Zool. Fenn., 1989, 26, 433. ant is the removal of soil ingestion as a route for the intake 29 L.H. J. Lajunen, Spectrochemical Analysis by Atomic Absorption and Emission, Royal Society of Chemistry, Cambridge, 1992. of metals. However, even allowing for an increased dietary 30 W. C. Ma, Arch. Environ. Contam. Toxicol., 1989, 18, 617. intake of metals similar to that modelled by Pascoe et al.,47 it 31 H. L. Alder and E.B. Roessler, Introduction to Probability and is still unlikely that wild animals could accumulate ecotoxicol- Statistics, Freeman, San Francisco, 6th edn., 1977. ogically significant levels of zinc, lead or cadmium from 32 M. S. Johnson, R. D. Roberts, M. Hutton andM. J. Inskip, Oikos, grassland colonising the tailings dam. 1978, 30, 153. 33 D. J. JeVries and M. C. French, Environ.Pollut., 1972, 3, 147. The outcome of this study contrasts with the risks to small 34 P. Williamson and P. R. Evans, Bull. Environ. Contam. Toxicol., mammals inhabiting older, disused metalliferous tailings dams, 1972, 8, 280. where the technology for metals recovery from crude ores was 35 W. C. Ma, W. Denneman and J. Faber, Arch. Environ. Contam. rudimentary and where residual metal concentrations in soils, Toxicol., 1991, 20, 266. vegetation and biota are much higher.32 The improved minerals 36 R. F. Shore and P. E. T. Douben, Ecotox. Environ. Safety, 1994, processing methods associated with modern mines, such as the 29, 101. 37 G. D. Kataev, J. Suomela and S. Palokangas, Oecologia, 1994, site which was the subject of this study, seems to provide 97, 491. equivalent environmental benefits to the much vaunted ‘clean 38 J. L. Domingo, J. Toxicol. Environ. Health, 1994, 42, 123. technology’ more usually associated with downstream indus- 39 G.J. Mutze, B. Green and K. Newgrain, Oecologia, 1991, 88, 529. trial operations in which metal concentrates are refined to 40 D. M. Gonzalespacheco, W. C. Buss, K. M. Koehler, W. F. finished goods. Woodside and S. S. Alpert, J. Nutr., 1993, 123, 90. 41 D. H. Hamer, Ann. Rev. Biochem., 1986, 55, 913. 42 P. Kille, A. Hemmings and E. A. Lunney, Biochem. Biophys. Acta, 1994, 12051. References 43 P. J. Aggett, in Zinc in Human Medicine, TIL, Isleworth, 1984, pp. 9–38. 1 B. A. Hunter, M. S. Johnson and D. J. Thompson, J. Appl. Ecol., 44 J. Chmeilnicka, T. Halatek and U. Jedlinska, Ecotox. Environ. 1987, 24, 601. Safety, 1989, 18, 268. 2 P. W. Purcell, M. J. Hynes and J. S. Fairley, Proc. Roy. Irish 45 S. C. Sheppard, Environ. Monit. Assess., 1994, 34, 27. Acad., 1992, 92B, 79. 46 W. N. Beyer, E. E. Connor and S. Gerould, J. Wildl. Man., 1994, 3 I. C. Boulton, J. A. Cooke and M. S. Johnson, J. Zool. (Lond.), 58, 375. 1994, 234, 409. 47 G. A. Pascoe, R. J. Blanchet and G. Linder, Arch. Environ. 4 B. A. Hunter, M. S. Johnson and D. J. Thompson, J. Appl. Ecol., Contam. Toxicol., 1994, 27, 44. 1989, 26, 89. 48 T. Johns and M. Duquette, Am. J. Clin. Nutr., 1991, 53, 448. 5 Z. Pucek and V. P. W. Lowe, in Small Mammals: Their 49 M. M. Vega, J. C. Solorzano, A. A. Medina, C. H. Luna and Productivity and Population Dynamics, ed. F. B. Golley, K. J. V. C. Salinas, Hum. Exp. Toxicol., 1996, 15, 872. Petrusewicz and L. Ryszkowski, Cambridge University Press, 50 F. Bronner and W. D. Stein, J. Nutr., 1995, 125, S1987. Cambridge, 1975, pp. 55–72. 6 R. E. Thomas and E. D. Bellis, J. Mamm., 1980, 61, 561. 7 W. J.Miller and M. W. Neathery, in Zinc in the Environment, Part Paper 9/01289C J. Environ. Monit., 1999, 1, 219–225 225

ISSN:0960-7919    

DOI:10.1039/a901289c

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

Cadmium concentrations in human kidney in the UK: 1978–1993 Thomas D. B. Lyon,a Elizabeth Aughey,b Roy Scottc and Gordon S. Fella aDepartment of Clinical Biochemistry, Glasgow Royal Infirmary University Trust, Glasgow, UK G4 0SF bDivision of Veterinary Anatomy, University of Glasgow, Veterinary School, Garscube, Stockiemuir Road, Glasgow, UK G61 1QH cDepartment of Urology, Glasgow Royal Infirmary University Trust, Glasgow, UK G4 0SF Received 18th February 1999, Accepted 3rd May 1999 Almost 2700 samples of human renal cortex have been collected from throughout the UK over a 16 year period from 1978 to 1993.The mean Cd concentration was 19 mg g-1 and the median 16 mg g-1.Smokers were, on average, about 5 mg g-1 higher than non-smokers. Cd increased from low concentration in the young to a maximum of 23 mg g-1 in middle age followed by a decrease in old age.Subjects who had died of renal disease had lower Cd concentrations. Geographical variations in the UK are small and the concentrations appear to be static over the 16 year period. Some 3.9% of the population had Cd concentrations >50 mg g-1, the critical level at which b2-microglobulin appears in urine.Aim of investigation Experimental procedure Pathologists from various parts of the UK agreed to provide Cadmium (Cd) is a toxic metal with a wide range of industrial samples of renal cortex (1–2 g wet weight) from the lower applications, which is now dispersed into the environment and pole of the left kidney taken at autopsy. The samples were can be found at low concentrations in most soils, plants and stored at -20 °C, in plastic containers without addition of animal species.Non-industrial human exposure is primarily preservative, and then forwarded frozen in dry cold to our from food and tobacco smoke. Occupational exposure to laboratory. The co-operating centres were Aberdeen, Bath, diVerent types of Cd compounds in fumes and dusts results in Belfast, Bristol, Chesterfield, Dumfries, Edinburgh, Glasgow, significant renal damage and bone disease.Monitoring of Kettering, Leicester, Leeds, London, Newcastle, SheYeld, industrial workers has yielded much information as to bio- Southampton and Watford. Information about age, sex, chemical markers of eVect.1 Cadmium is a persistent toxin underlying cause of death and smoking habit was requested. with a biological half-life of over 20 years.The main target organ is the kidney, which contains about 1/3 of the whole body content. Concerns as to the eVects on the health of the Analysis and quality control general population from environmental pollution were raised The samples were thawed and the wet weight established initially by the Japanese experience of severe bone disease before oven drying overnight at 80 °C.The cadmium concenrelated to Cd release into agricultural land in the waste water trations are reported on a wet weight basis as this is the most from zinc refineries and its uptake by the rice crop.1 In the widely used convention (multiply by 5 to convert to dry UK a geochemical soil survey revealed that high amounts of weight).The dried tissue was digested in 50 ml Pyrex tubes Cd and other metal residues were present in land now used with 7 ml of AristarR grade nitric acid in a heating block for for crop production. This led to an extensive investigation2 60 min at 110 °C and the digest made up to 25 ml with which concluded that there was no evidence of an adverse deionized water.The digests were stored in sealed plastic health eVect to the local population. bottles at 4–10 °C. More recently, Lauwerys et al.3 (Cadimbel Study) have The cadmium concentration of the digest was measured by reported on the renal eVects of Cd on Belgian populations flame atomic absorption spectrometry (FAAS) using a Perkinliving in areas where heavily polluted soils have been identified.Elmer 3030 with air–acetylene flame using the cadmium They have concluded that some of the general population of 228.8 nm line. Belgium have absorbed suYcient Cd to cause slight renal The accuracy of this method was established by dysfunction. They base this on the finding that urine Cd Cunningham5 by comparison with a flame fluorescence excretion above 2 mg l-1 was associated with an increased technique and replicate analysis (n=4) of the standard referurinary excretion of b-microglobulin and other biochemical ence material (National Bureau of Standards 1577, Bovine markers of renal tubular damage.They have calculated that Liver). Cadmium in bovine liver reference material was deterthis urinary output of Cd is equivalent to a renal cortex Cd mined to be 0.26±0.04 mg g-1 (the certified value is content of 50 mg g-1 wet weight. 0.27±0.04 mg g-1). The Bovine Liver SRM has a relatively We now report on the determination of some 2700 renal low cadmium concentration and was not suitable for use as a cortical Cd contents of human kidney obtained at autopsy quality control material in this study. Samples of freeze dried from sixteen locations throughout the UK and conducted over horse kidney with an assigned value of 224 mg g-1 were a 16 year period.The data presented here is an inclusive obtained from the Karolinska Institute and analysed with each extension of that described by Scott et al.,4 which covered the batch of human kidney. Subsequently, IAEA H-8 horse kidney with an assigned value of 184–193 mg g-1 dry weight (95% period 1978–1986.J. Environ. Monit., 1999, 1, 227–231 227Table 1 Distribution of subjects by age (in decades) and gender Age M F All 1 1 1 2 2 3 3 6 3 15 11 26 4 17 20 37 5 51 37 88 6 135 78 213 7 279 231 510 8 371 313 684 9 213 316 529 All 1085 1010 2095 Not known 620 Fig. 1 Quality control chart for IAEA Horse Kidney from 1986 to 1993. decades. The frequency distribution for cadmium in renal cortex was skewed.However, the distribution of the log transformed data was normal and so all statistical tests were confidence interval ) was used from 1986 onwards. The quality performed using the log-normal data. The error bars on all control chart showing the mean value for the IAEA horse figures are 95% confidence intervals. DiVerences between sev- kidney from 1986 to 1993 is shown in Fig. 1. The average eral group means were tested for by one-way analysis of mean over the period was 193±7.9 (s). There was no temporal variance. The Tukey pairwise test was used for ‘follow on’ trend and no results were out of bounds (±3 s). analysis. The association between cadmium concentration and During the course of this study inductively coupled plasma age, sex and smoking habit was examined by multiple mass spectrometry (ICP-MS) became available and an analytregression analysis.ical comparison was made between the FAAS method and All statistical calculations were performed using MinitabR ICP-MS. The results are shown in Fig. 2. The diVerences 10 Xtra. between the values obtained by FAAS and ICP-MS for a number of kidney samples are plotted against the mean result as suggested by Bland and Altman.6 The data are normally Results and discussion distributed about zero diVerence, indicating that there is no Cadmium concentration bias between the two methods.The limit of agreement is +3.2 to -4.4 mg g-1. The frequency distribution of cadmium concentrations in renal cortex is shown in Fig. 3. The distribution has a mean of Population characteristics 19.1 mg g-1 (n=2659) and is skewed. The median is 16 and the 95% confidence interval is 15–16 mg g-1. There is a con- Kidney samples (n=2659) from the general population, siderable variance, with a range of 0.1–163 mg g-1 and inter- including three known coppersmiths, were collected over a 16 year period from 1978 to 1993.The majority of samples were from subjects who had died of natural causes, although some did come from accident victims. The distribution of subjects by age (in decades) and sex is shown in Table 1. In the group of subjects who smoked (or were ex-smokers) a higher proportion were men (60%) compared with women (40%). Subject information is incomplete and there is missing data on gender (n=451, 17%), age (n=620, 23%) and smoking habit (n= 1708, 64%).Statistical analysis For statistical analysis former smokers were grouped along with the smokers and the ages at death were grouped into Fig. 2 Plot of the diVerence (ICP-MS-FAAS) against the mean values for a set of 23 kidney cortexes. The dashed lines indicate the limits of agreement. The solid lines delineate the 95% confidence Fig. 3 Descriptive statistics output from Minitab 10 Xtra for 2659 interval of the mean diVerence and as this includes 0 there is no bias between the methods. kidney cortex Cd concentrations (mg g-1) from the UK. 228 J. Environ. Monit., 1999, 1, 227–231quartile range 9-25 mg g-1. Three of the subjects who were known coppersmiths7 (and therefore exposed to cadmium) had high concentrations of 55, 142 and 163 mg g-1.The mean concentrations found are higher than those of 1–2 mg g-1 found in kidney from the nineteenth century and earlier in the twentieth century.8,9 The present mean is below the ‘critical level’ of 200 mg g-1 suggested by the FAO/WHO10 as the concentration at which the first signs of renal damage, that is selective proteinuria, will become apparent.The Cadimbel Study, however, has suggested that slight renal dysfunction will be seen in 10% of a general population when the urine excretion of Cd exceeds 2 mg l-1, which is observed when the renal cortex Cd is >50 mg g-1. Elinder11 has collated arithmetic means for renal cortex cadmium concentrations from a number of countries for subjects in the age range 40–59 years.We found an arithmetic mean of 23 mg g-1 for this age group in the UK, which is similar to the findings of for Finland, France, USA, Sweden Fig. 5 Frequency distribution of Cd concentration (mg g-1) in cortex and Austria (range 20–40 mg g-1). The range of arithmetic of smokers and non-smokers. For purposes of comparison the distri- means in Japan is much higher, 60–140 mg g-1.bution for the non-smokers has been scaled so as to match the area for the smokers distribution. Sex The geometric mean value for males (n=1202) was 14.8 mg g-1 from both Sweden and the USA,15 but these studies involved and for females (n=1089) 14.6 mg g-1. A ‘t ‘ test performed relatively small numbers (n=74 and n=54), respectively. Other on the lognormal data indicates no significant diVerence ( p= causes for the discrepancy are possible diVerences in the 0.48) between males and females.Neither were there any cadmium content of cigarettes and in diVerences in the selection diVerences between males and females across the age groups. of subjects. These possibilities and also the extent of the smoking habit on cadmium cortex concentrations have been Age discussed elswhere.4 The association of cadmium concentrations with age for 2122 Cause of death individuals is shown in Fig. 4. Cadmium concentrations rise from the small amounts found in the youngest children to a The seven underlying causes of death were used to group the maximum at age 40–60 years. This trend has been noted by data and a one-way analysis of variance on the log-normalised Elinder.11 Infants are known to have low cadmium concen- cadmium concentrations was performed.The analysis indicates trations in their tissues12,13 and it seems reasonable to infer that there are significant diVerences ( p<0.05) between the that cadmium accumulates in kidney throughout life to a group means. The means and confidence intervals are shown maximum of 23 mg g-1 at 50–60 years of age.The subsequent in Fig. 6. The renal disease group is markedly lower, geometric fall in old age cannot be explained by changes in the environ- mean 8 mg g-1, than the other groups, but the size of this mental exposure of the elderly.14 However, it is known that group is small (n=13). Nevertheless, Tukey’s pairwise comthe mass of the kidney decreases in old age with an associated parisons suggest that the diVerence is significant.The mean decrease in renal function. values of all of the other groups are all outwith the confidence interval for the renal group. Likewise, the miscellaneous group Smoking habits is lower than the pulmonary, cancer, heart and cerebrovascular groups. It has been suggested that cadmium may be a causative The subjects were divided into smokers (n=662, including factor in hypertension,16 cardiovascular disease17 and in vari- pipe, cigar and former smokers) and non-smokers (n=372).ous cancers;18–21 however, no evidence of elevated cadmium The results are shown as a histogram in Fig. 5. The nonin tissue from those dying from cardiovascular disease, cerebro- smokers had a median renal cadmium concentration of vascular incident, cancer or bronchitis has been found.22–24 13 mg g-1 (geometric mean=12.6), which is significantly lower than that of the smokers 18 mg g-1 (geometric mean=16.4).The diVerence in geometric means, 3.8, is not as large as the diVerence between geometric means of ~10 mg g-1 reported Fig. 6 Variation of corticular Cd concentration (mg g-1) with Fig. 4 Distribution of Cd concentration (mg g-1) as a function of age (note log scale). underlying cause of death (note log scale). J. Environ. Monit., 1999, 1, 227–231 229from Uppsala with earlier published work for Stockholm and found that the cadmium concentrations had reduced to about 40% of the 1976 concentrations.25,26 This was an unexpected finding as environmental contamination in Sweden has risen over the same period.Unfortunately, an unequivocal interpretation of the Swedish results is not possible in the absence of intercomparative data for the two studies carried out there. Multiple regression analysis A multiple regression model using ‘age’, ‘smoking habit’ and ‘sex’ as explanatory variables was fitted to the data. Although the coeYcients for ‘age’ and ‘smoking’ were highly significant the model was poor and could only explain about 8% of the variation leaving 92% unexplained.Morgan and Sherlock27 demonstrated a strong relation between average dietary intake and kidney cadmium content so it is reasonable to infer that individual dietary intake and associated absorption and Fig. 7 Variation of corticular Cd concentration (mg g-1) with excretion of cadmium is the principal cause of the observed geographical locality (note log scale). variation.Our results suggest that the cadmium concentrations in heart Association of cadmium and zinc disease and in cancer, geometric mean 15 mg g-1, are slightly higher than the miscellaneous group, geometric mean Human autopsy studies have shown an association between 12.6 mg g-1, but the diVerence is not statistically significant.cadmium and zinc.28–30 We confirm a weak association and found that the regression of cadmium on zinc explained 24% Geographical variations within the UK of the variance. A regression of log (Cd) on log (Zn) explained 36% of the variance. The means and confidence intervals for the lognormalised data were calculated for the sixteen localities and the results are shown in Fig. 7. One-way analysis of variance indicates that Conclusions there are statistically significant diVerences ( p<0.003) between This was a large study involving nearly 2700 kidney cortex the diVerent localities. Only one pairwise diVerence was found analyses performed over a 16 year period. Good quality to be significant; Glasgow was slightly higher (geometric control of the analyses was maintained throughout the dur- mean=15.5) than Chesterfield (geometric mean=13.5).The ation of the project. The study confirms the results of smaller diVerence can possibly be explained by the age profile of the studies in other countries which demonstrated a relationship two groups; the proportion of subjects of 80+ years (i.e., with between kidney cortex cadmium and age and smoking habit.age lowered cadmium) is greater in Chesterfield (35%) than in It also shows that the UK levels are among the lowest found. Glasgow (21%). Also, a higher prevalence of smoking in Geographical variations within the UK have been shown to Glasgow is likely and may contribute to the diVerence (smokbe small.No change over the 16 years of the study was ing habit figures for Chesterfield were not available). In general apparent, suggesting cadmium uptake by the UK population though, geographical variations within the UK appear to is not increasing. However, if the critical threshold of 50 mg g-1 be small. in renal cortex, as advocated by Lauwerys et al.,3 is accepted, then a mean value of 19 mg g-1 is not suYciently removed Temporal variations from the critical threshold.The FAO/WHO10 suggested ‘criti- Since small geographical variations may exist the possibility cal level’ of 200 mg g-1 is a factor of ten greater than the mean of any temporal trend was investigated using only data from value found here. If the ‘critical value’ is reduced to 50 mg g-1, the Glasgow area.The data are plotted in Fig. 8. No adjust- which is only twice the mean value, then about 3.9% of the ment was made for any age diVerences between year groups. population would exceed this threshold; any further exposure One-way analysis of variance indicates that there are diVer- of the UK population would therefore be undesirable. ences between the years (F=2.83, p=0.001) but there is no conclusive evidence of a trend with time.However, if the 1990 Acknowledgments data is omitted there is a suggestion of a slight rise. Friis et al. have compared recent (1996) kidney cortex concentrations This study was carried out under a contract (PECD/7/8/36) from the Department of the Environment. The authors thank Mrs. Mary Reilly and Mrs.Lana MacDonald of the Veterinary School, Glasgow, for expert technical assistance. The co-operation of those who supplied the kidneys from post mortem examinations and those who provided demographic information is greatly appreciated. References 1 Cadmium and Health: a Toxicological and Epidemiological Appraisal, eds. L. Friberg, C. G. Elinder, T. Kjellstrom and G. F. Nordberg, CRC Press, Boca Raton, Florida, USA, 1985, vol. 2. 2 J. C. Sherlock, G. A. Smart, B. Walters, W. H. Evans, Fig. 8 Temporal variation of corticular Cd concentration (mg g-1) for D. J. McWeeny and W. Cassidy, Sci. Tot. Environ., 1983, 29, 121. 3 R. Lauwerys, A. Bernard, J. P. Buchet, H. Roels, P. Bruaux, Glasgow (note log scale). 230 J. Environ. Monit., 1999, 1, 227–231F. Claeys, G. DucoVre, P.De Plaen, J. Staessen, A. Amery, 16 H. Schroeder, J. Chron. Dis., 1965, 18, 647. 17 G. L. Friberg, M. Piscator, G. F. Nordberg and T. Kjellstrom, R. Fagard, P. Lijnen, L. Thijs, D. Rondia, F. Sartor, A. Saint Cadmium in the Environment, CRC Press, Cleveland, Ohio, 2nd Remy and L. Nick, Acta Clin. Belg., 1991, 46, 219. edn, 1974, pp. 116–119. 4 R. Scott, E. Aughey, G. S.Fell and H. J. Quinn, Human Toxicol., 18 C. L. Potts, Ann. Occup. Hyg., 1965, 8, 55. 1987, 6, 111. 19 J. M. Morgan, Cancer, 1970, 25, 1394. 5 C. Cunningham, FIMLS thesis, Glasgow Royal Infimary, 1980. 20 L. N. Kolonel, Cancer, 1976, 37, 1782. 6 J. M. Bland and D. G. Altman, Lancet, 1986, i, 307. 21 T. Kjellstrom, L. Friberg and B. Rahnster, Environ. Health 7 R. Scott, P. J. Paterson, E.A. Mills, A. McKirdy, G. S. Fell, Perspect., 1979, 28, 199. J. M. Ottaway, F. E. R. Husain, O. P. Fitzgerald-Finch, 22 T. L. M. Syversen, T. K. Stray, G. B. Syversen and J. Ofstad, A. J. Yates and A. Lamont, Lancet, 1976, 21, 396. Scand. J. Clin. Invest., 1976, 36, 251. 8 C. G. Elinder and T. Kjellstrom, Ambio, 1977, 6, 270. 23 J. M. Morgan, Arch. Intern. Med., 1969, 123, 405. 9 G. A. Drasch, Sci. Tot. Environ., 1983, 26, 111. 24 G. P. Lewis, W. J. Jusko, L. L. Coughlin and S. Hartz, J. Chron. 10 FAO/WHO Evaluation of Certain Food Additives and the Dis., 1972, 25, 717. Contaminants Mercury, Lead and Cadmium. Sixteenth Report of 25 L. Friis, L. Petersson and C. Edling, Environ. Health Perspect., the Joint FAO/WHO Expert Committee on Food Additives, 1998, 106, 175. Geneva, Switzerland, 1974. 26 C. G. Elinder, T. Kjellstrom, L. Friberg, B. Lind and L. Linnman, 11 C. G. Elinder, in Cadmium and Health: A Toxicological and Arch. Environ. Health, 1976, 31, 292. Epidemiological Appraisal, eds. L. Friberg, C. G. Elinder, 27 H. Morgan and J. C. Sherlock, Food Add. Contam., 1984, 1, 45. T. Kjellstrom and G. F. Nordberg, CRC Press, Boca Raton, 28 C. G. Elinder, M. Piscator and L. Linnman, Environ. Res. 1977, Florida, USA, 1985, vol. 1, pp. 81–102. 13, 432. 12 M.Piscator, VDI Berichte, 1975, 203, 43. 29 D. I. Hammer, A. V. Colucci, V. Hasselblad, M. E. Williams and 13 S. B. Gross, D. W. Yeager and D. S. Middendorf, J. Toxicol. C. J. Pinkerton, Occup. Med., 1973, 12, 956. 30 M. Piscator and B. Lind, Arch. Environ. Health, 1972, 24, 426. Environ. Health, 1976, 2, 153. 14 C. C. Travis and A. G. Haddock, Environ. Res., 1980, 22, 46. 15 T. Kjellstrom, Environ. Health Perspect., 1979, 28, 169. Paper 9/01366K J. Environ. Monit., 1999, 1, 227–231 231

ISSN:0960-7919    

DOI:10.1039/a901366k

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

QUASIMEME laboratory performance study of the biological eVects of tributyltin (imposex and intersex) on two marine gastropod molluscs† I. M. Davies,*a A. Minchin,a B. Bauer,b M. J. H. Hardinga and D. E.Wellsa aFRS Marine Laboratory, PO Box 101, Victoria Road, Torry, Aberdeen, UK AB11 9DB. E-mail: daviesim@marlab.ac.uk, minchina@marlab.ac.uk, hardingm@marlab.ac.uk, wells@marlab.ac.uk; Fax:+44 1224 295511 bInstitut fur Spezielle Zoologie and Vergleichende Embriologie, HueVerstrasse 1, D 48149, Muenster, Germany Received 8th April 1999, Accepted 27th April 1999 The disruption of the endocrine systems of gastropod molluscs and consequential physiological changes (imposex and intersex) are used as biomarkers for environmental contamination by tributyltin compounds.The first international laboratory performance study on the determination of imposex and intersex in neogastropod molluscs, Nucella lapillus and Littorina littorea has been undertaken by the QUASIMEME (Quality Assurance of Information for Marine Environmental Monitoring in Europe) project.Samples of live gastropods were distributed and participants were asked to record shell height and sex, together with penis length and vas deferens sequence stage (VDS) in Nucella or the intersex stage (IS) and prostate length in Littorina.Calculations were made of vas deferens sequence index (VDSI) and the relative penis size index (RPSI) in Nucella and of intersex stage index (ISI ) and the average female prostate length (FPrL) in Littorina. Thirteen (87%) of the 15 participating laboratories returned data.The remaining two laboratories asked to participate in later exercises. For Nucella, seven laboratories reported sex ratios significantly diVerent from the reference laboratory data. DiVerences in penis length measurements between laboratories were largely random, although there were indications of systematic errors aVecting the data from three laboratories.Seven laboratories reported satisfactory data (Z-score |Z|<2) for VDSI. The inclusion of a high proportion of sub-adults in the Nucella samples may have made separation of the sexes more diYcult than in mature adults. The sub-adults will have had smaller pene than mature adults in the same population, and therefore any errors (random or systematic) in the measurement of penis length or observation of reproductive organs would have a potentially greater impact on the final reported values of the summary imposex indices.The Littorina sample did not show a high degree of intersex (ISI=0.41). The laboratories could determine the sex of Littorina reliably and only one laboratory reported data significantly diVerent from the reference laboratory.All except two laboratories reported satisfactory data for ISI. Introduction the Sea (ICES). A Training Workshop on the measurement It has been recognised for some years that the presence of of imposex and intersex was held at the FRS (Fisheries tributyltin (TBT) compounds in the marine environment, Research Services) Marine Laboratory, Aberdeen primarily derived from antifoulants, can lead to disruption (24–26 September, 1997) with the dual purposes of providing of the endocrine systems and characteristic physiological training in measurement techniques and to review and revise responses in gastropod molluscs.1,2 These alterations to the the JAMP Guideline document.10 Participants in the workshop female genital systems are termed imposex [superimposition recognised the need for a series of international laboratory of penis and/or vas deferens on female prosobranch snails, performance studies of the determination of imposex and notably the dogwhelk Nucella lapillus (L.)] and intersex (patho- intersex in marine snails, using the revised JAMP Guideline.logical alterations in the oviduct of littorinids and replacement The QUASIMEME (Quality Assurance of Information for of female by male organs), have proved to be sensitive Marine Environmental Monitoring in Europe) project provides biomarkers for the degree of organotin (especially TBT) a series of laboratory performance studies for chemical pollution in coastal waters.3–6 measurements in marine matrices.11 Part of the programme The inclusion of the biological eVects of tributyltin has developed towards providing quality assurance (QA) for compounds in Nucella lapillus and Littorina littorea (imposex measurements of polyaromatic hydrocarbon (PAH) metaband intersex respectively7,8) in the international marine Joint olites and organotin compounds in water and biota with a Assessment and Monitoring Programme (JAMP) of the Oslo view towards linking chemical measurements and biological and Paris Commission (OSPAR) immediately raised the need eVects studies.Conveners of the workshop collaborated with for quality assurance procedures including standardisation of the QUASIMEME project in the organisation of the first analytical techniques, training of scientists, and interlaboratory international exercise of this nature for the biological eVects studies.9 Detailed guidelines for sample site selection, sample of tributyltin compounds.12,13 This report presents an account collection, storage and data reporting were developed by of the exercise and of the lessons learned for future exercises.working groups under the OSPAR system, incorporating advice from the International Council for the Exploration of Materials and methods Samples of Nucella lapillus (sample code Q15001BT) and Littorina littorea (Q15002BT) were prepared and supplied by †©Crown copyright. J.Environ. Monit., 1999, 1, 233–238 233the FRS Marine Laboratory, Aberdeen. Nucella were collected described.3,8,14,15 Two reference laboratories were used in this study. Nucella were analysed by the FRS Marine Laboratory, from Skatey Shore, Stonehaven on 30 January and Littorina from Aberdeen harbour entrance on 2 February, 1998.The Aberdeen, Scotland and Littorina by the Institut fu� r Spezielle Zoologie und Vergleichende Embryologie, Muenster, snails were stored at the FRS laboratory aquarium until the morning of 3 February, the day of dispatch. The bulk samples Germany. These laboratories examined 424 Nucella and 241 Littorina respectively.of snails were placed on a tray, coned and quartered into samples of 50+, enclosed in nylon mesh bags, and placed into Styrofoam boxes lined with paper. Additional paper was added Results as packing material to minimise the movement of the animals The gastropod samples were generally received at UK during transit. A domestic freezer ice pack was placed on top destinations on 4 February and outside the UK on of the paper packing to keep the snails cool and the boxes 5/6 February and all these were live and in good condition.were sealed. All samples were packaged on the day of dispatch. Some samples suVered long delays, e.g., by customs authorities, Samples were distributed by overnight post (UK) or courier.and replacement samples were sent on 19 February. Participants in the exercise were provided with the samples of snails, the protocol for the exercise, the JAMP Guidelines Imposex in Nucella lapillus for Contaminant-specific Biological EVects Monitoring for TBT, as amended at the TBT EVects Training Workshop, Participants recorded their data on the data collector diskette September, 1997 and the data collector diskettes with from which the information (Table 2) was transferred directly instructions for their installation and use.10 by the QUASIMEME project oYce to the QUASIMEME The protocol for the exercise stated that: ‘This study covers data base.The initial step in data assessment was to check the the determination of imposex in Nucella lapillus (QIS001BT) calculations of the summary parameters [relative penis size and intersex in Littorina littorea (QIS002BT).Forty snails index (RPSI) and vas deferens sequence index (VDSI)] using should be chosen at random and examined from each sample, the formulae in the OSPAR guidelines. In some cases, particiand the observations on each individual snail should be pating laboratories had made errors in calculations.The reported using the QUASIMEME data collector and the datae-calculated values for each laboratory (Table 2) were used sheets included with this protocol. The observations listed in subsequent data analysis. below (Table 1) should be made, following the enclosed OSPAR JAMP guidelines. Nucella—homogeneity of samples. All laboratories reported Samples should be stored cool prior to analysis, preferably individual shell height measurements for Nucella and these in sea water, but it is advisable to analyse the samples as soon measurements (Fig. 1) were used to assess the homogeneity of as possible after receipt. You are requested to carry out the the samples. Visual examination of data indicated that two of observations following Technical Annex 3 of the OSPAR the participating laboratories (Q327P, Q158P) had reported Guidelines for Biological EVects Monitoring particularly for lower values for shell height than the other laboratories, and stage descriptions.The guidelines indicate that Nucella should this was confirmed by analysis of variance (ANOVA). The be examined live without narcotisation and that Littorina ANOVA was repeated omitting these two samples and indishould be narcotised with 7% magnesium chloride solution.cated that significant diVerences ( p<0.05) still existed between Parasitised animals should be discarded and alternative snails samples. The mean shell height of the remaining samples selected until a sample size of 40 has been obtained’. ranged from 22.79 to 24.60 mm (Table 2) and the ‘between The OSPAR Guidelines require that assessments of imposex sample’ sum of squares was 7% of the ‘within sample’ sum of and intersex be carried out according to the methods squares.The organisers decided that the remaining diVerences between these samples were small and would not compromise the exercise. Table 1 Measurements requested for the determination of imposex (Nucella lapillus) and intersex (Littorina littorea)a Nucella—penis length.Since there were significant diVerences in shell height between samples, the data for penis length for Nucella lapillus (QIS001BT) Shell height/mm both males and females were examined in relation to shell Sex coded (male=1, female=0) height. In both cases a positive correlation (Fig. 2) was found Penis length (eye piece graticule units) between mean penis length and the mean shell height. In order Penis length/mm to make comparisons between samples of diVerent mean shell Vas deferens stage height it was therefore necessary to correct mean penis length Calculated indices: Vas deferens stage index (VDSI) measurements to a standard shell height. Regression lines were Relative penis size index (RPSI) fitted to the relationships between penis length and shell height Littorina littorea (QIS002BT) Shell height/mm and used to correct the mean penis length measurements Sex coded (male=1, female=0) (Table 3) to the mean shell height (23.26 mm) for all Prostate length (graticule units) laboratories (omitting laboratories Q327P and Q158P).Prostate length/mm A bi-plot (Fig. 3) of the mean penis length for males and Intersex stage Calculated indices: Intersex stage index (ISI ) females (corrected for shell height) for all laboratories suggests Average female prostate length that diVerences between most laboratories arise from random (FPrL) errors, however there are indications that some degree of aThe indices referred to in Table 1 are calculated as follows: Vas systemic error has aVected the data for three (Q154P, Q158P deferens sequence index (VDSI) in Nucella=sum of imposex stage and Q104P) of the participating laboratories.values of all females sampled/number of females. Relative penis size index (RPSI) in Nucella=[(cube of average length of female Nucella—sex identification. The reference laboratory penis)/(cube of average length of male penis)]×100%.Intersex index examined 424 individual Nucella from the bulk sample. The (ISI ) in Littorina=sum of intersex stage values of all females sampled/ number of females. Average female prostate length (FPrL) in proportions of males and females found (47.2 and 52.8% Littorina=sum of prostate lengths (mm) of all females sampled/ respectively) were taken as the best estimates of the sex number of females.In addition, the relative penis length index (RPLI) composition of the bulk sample. There was no significant in Nucella was calculated as [(average length of female penis)/(average diVerence between the mean shell heights of males and females. length of male penis)]×100%. The numbers of males reported by the participating labora- 234 J.Environ. Monit., 1999, 1, 233–238Table 2 Summary of data received from participating laboratories and calculations undertaken by QUASIMEME for Nucella lapillusa Mean penis lengthb/mm Mean shell Recalculated RPSI Recalculated heightb/mm Male Female VDSI VDSIb (%) RPSIb (%) Q104P 22.79 2.95 1.91 3.95 3.95 27.14 27.14 Q128P 24.36 2.47 1.35 3.46 3.46 26.20 16.18 Q131P 24.57 2.46 1.31 4.00 4.00 15.10 15.10 Q136P 23.70 2.22 1.47 3.92 3.92 3.00 29.19 Q137P 22.87 2.27 1.50 3.95 3.95 28.57 28.57 Q154P 23.92 2.88 1.82c 2.70 2.70 16.80 25.22 Q156P 23.98 2.46 1.77 2.67 2.67 37.21 37.41 Q158P 20.57 1.14 0.65 4.00 4.00 18.80 18.80 Q247P 24.60 2.18 1.79 —d 3.23 —d 55.36 Q327P 19.53 1.48 0.86 1.81 1.81 19.60 19.60 Q328P 23.69 1.95 1.64 1.70 2.57 1.88 59.52 Q329P 24.47 2.38 1.37 3.95 3.95 19.07 19.15 Q330P 24.37 2.69 1.42 3.90 3.90 14.84 14.74 aFigures in bold indicate that calculation errors were made by the participating laboratories.bCalculations by QUASIMEME. cConversion from graticule units was miscalculated by laboratory. dNo calculated values were received for this index. Fig. 2 Mean penis length for males (a) and females (b) as a function of shell height. 52–66%, RPSI 14–29%) than the values for the other laboratories (RPLI 52–85%, RPSI 14–61%). Fig. 1 Box and whisker plots of shell height for Nucella lapillus (a) Nucella—vas deferens sequence index (VDSI). The reference and Littorina littorea (b). laboratory examined 224 female Nucella and obtained an overall VDSI value of 3.8. Based on these data, the predicted standard error of the VDSI on samples of 40 snails is 0.1 VDS tories in their sub-samples of 40 individuals ranged from 19 (47.5%) to 39 (97.5%).Chi-square statistics indicated that the units. The individual laboratory data may be interpreted in terms of Z-scores, following standard ISO/IUPAC16 data from seven (53.8%) laboratories were significantly diVerent from the reference laboratory data suggesting that some procedures where: misidentification of females as males had occurred at these Z=(observed value-target value)/total allowable error laboratories.The data in Table 3 are ordered by the proportion of Nucella identified as males. The application of this procedure within QUASIMEME is detailed by Wells and Cofino.17 If 0.1 VDS units is taken as The data for penis length and summary parameters were examined in two groups according to whether the sex ratio the total allowable error, then seven laboratories (Q158P, Q131P, Q136P, Q137P, Q329P, Q330P and Q104P) reported was significantly diVerent from the target value.T-tests indicated that there were no diVerences in male or female mean satisfactory data, i.e., -2<Z<2.The remaining six laboratories reported unsatisfactory data for VDSI. The OSPAR penis lengths between the two groups. However the calculated values of relative penis length index (RPLI) and RPSI for the Guideline (OSPAR 1997) indicates that in field surveys, VDSI values estimated by interpolation between sampling stations group of laboratories which had not shown significant diVerences from the reference laboratory were less variable (RPLI should be within 0.5 units of the true value with 90% confidence J.Environ. Monit., 1999, 1, 233–238 235Table 3 The recalculated summary values for Nucella lapillus that were used in data analysis, ordered by the proportion of snails identified as males Sex Mean penis length/mm Mean shell RPLI RPSI Male Female Chi-square height/mm Male Female VDSI (%) (%) Q158P 39 1 <0.001 20.57 1.59 0.97 4.00 83.68 58.60 Q154P 31 9 0.001 23.92 1.68 1.03 2.70 55.03 16.67 Q156P 31 9 0.001 23.98 2.77 1.52 2.67 72.12 37.52 Q131P 27 13 0.010 24.57 2.24 1.16 4.00 51.73 13.84 Q128P 27 13 0.010 24.36 2.28 1.22 3.46 53.52 15.33 Q136P 27 13 0.010 23.70 2.14 1.42 3.92 66.44 29.33 Q247P 26 14 0.024 24.60 2.34 1.69 3.23 84.70 60.76 Q327P 24 16 0.104 19.53 1.95 1.63 1.81 56.51 18.06 Q137P 20 20 0.720 22.87 2.34 1.55 3.95 66.15 28.95 Q329P 19 21 0.967 24.47 2.17 1.23 3.95 60.73 22.40 Q330P 19 21 0.967 24.37 2.50 1.29 3.90 61.27 23.00 Q104P 19 21 0.967 22.79 3.03 1.96 3.95 64.52 26.86 Q328P 19 21 0.967 23.69 2.12 1.30 2.57 51.60 13.74 the formulae as outlined in the revised (1997) JAMP guidelines.As for Nucella, some calculations for Littorina summary parameters showed errors and the re-calculated values are shown in Table 4. Littorina—homogeneity of samples. As for Nucella, the individual shell height measurements for Littorina were used to assess the homogeneity of the samples. Visual examination of data (Fig. 1) suggested that the two participating laboratories which had reported low values for shell height in Nucella (Q327P, Q158P) again reported lower shell heights compared to the other laboratories.This was confirmed by ANOVA. Fig. 3 Bi-plot of mean penis lengths (corrected for shell height) for The ANOVA was repeated omitting these two samples and male and female Nucella lapillus. indicated that significant diVerences still existed between samples.The mean shell height of these samples ranged from in the VDSI range 2–6. Depending upon the sampling density 19.13 to 20.37 mm (Table 4). The ‘between sample’ sum of and the gradient of eVect, this may equate to a less demanding squares was 5% of the ‘within sample’ sum of squares. The value for the total allowable error. For example, if 0.25 VDS organisers decided that the remaining diVerences between these units is taken as the total allowable error, then the data from samples were small and would not compromise the exercise.one further laboratory (Q128P) would be assessed as satisfactory and from four laboratories as unsatisfactory. Variation of the total allowable error between 0.1 and 0.25 units in this Littorina—sex identification.The reference laboratory exercise does not greatly alter the assessments of individual examined 241 individual Littorina from the bulk sample. The laboratory performance. proportion of males and females found (44.8 and 55.2% respectively) were taken as the best estimates of the sex Intersex in Littorina littorea composition of this bulk sample. There was no significant diVerence between the shell heights of males and females. The All data received were entered into the QUASIMEME data base and are summarised in Table 4.The initial step in the numbers of males reported by the participating laboratories, in their sub-samples of 40 individuals ranged from 12 (30%) data assessment was to re-calculate the summary parameters [interstage sex (ISI ) and female prostate length (FPrL)] using to 26 (65%).Chi-square statistics indicated that the data from Table 4 Summary of data received from participating laboratories and calculations undertaken by QUASIMEME for Littorina littoreaa Sexb Mean shell height/mm Recalculated Recalculated Male Female Chi-squareb Shell heightb ISI ISIb FPrL FPrLb Q104P 21 19 0.3290 19.13 0.63 0.63 Q128P 19 21 0.7337 19.82 0.24 0.24 0.14 Q131P 16 24 0.5395 19.40 0.04 0.04 Q136P 18 22 0.9822 20.37 0.23 0.23 Q137P 16 24 0.5395 18.72 0.04 0.04 Q154P 16 24 0.5395 19.73 0.75 0.75 0.76 0.76 Q156P 15 25 0.3516 19.75 0.72 1.00 Q158P 26 14 0.0103 16.33 1.50 1.50 0.13 0.13 Q247P —c —c —c —c —c —c —c —c Q327P 23 17 0.1070 15.14 0.12 0.12 Q328P 18 22 0.9822 19.66 0.44 0.46 Q329P —c —c —c —c —c —c —c —c Q330P 12 28 0.0594 19.67 0.75 0.72 aFigures in bold indicate that calculation errors were made by the participating laboratories.bCalculations by QUASIMEME. cNo data received for Littorina. 236 J. Environ. Monit., 1999, 1, 233–238Table 5 List of laboratories participating in the 1998 QUASIMEME laboratory performance study for imposex and intersex in gastropod molluscs Halifax Fisheries Research Fisheries and Oceans Canada 1707 and 1727 Lower Nova Scotia B3J 2S7 Canada Laboratory Water Street Halifax National Environmental Research PO Box 358 Frederiksborgvej 399 DK-4000 Denmark Institute Institut fur Spezielle Zoologie und HueVerstrasse 1 D 48149 Muenster Germany Vergleichende Embriologie Limnomar Bei der Neuen Munze 11 D-22145 Hamburg Germany Institute of Biology University of Iceland Grensasvegi 12 108 Reykjavik Iceland Fisheries Research Centre Marine Abbotstown Castleknock Dublin 15 Ireland Institute Martin Ryan Science Institute National University of Ireland Galway Ireland Netherlands Institute for Sea PO Box 59 1790 AB Den Burg Texel Netherlands Research Norwegian Institute for Water PO Box 173 Kjelsaas 0411 Oslo Norway Research University of Algarve Campus de Gambelas 8000 Faro Portugal Fisheries Research Services, Marine P O Box 101 Victoria Road Aberdeen AB11 9DB UK Laboratory Scottish Environment Protection Heriot-Watt Research Park Avenue North Riccarton Edinburgh EH14 4AP UK Agency (East), Clearwater House Scottish Environment Protection 5 Redwood Crescent Peel Park East Kilbride Glasgow G74 5PP UK Agency (West) Industrial Research and Technology 17 Antrim Road Lisburn Co Antrim BT28 3AL UK Unit Environment Agency-North East Skinnerburn Road Newcastle-upon-Tyne Tyne and Wear NE4 7AR UK only one (9%) laboratory (Q158P) were significantly diVerent The exercise was made more diYcult by collecting Nucella during February/March. In the east of Scotland, adult Nucella from the reference laboratory data.are normally concealed in deep fissures between rocks at this time of year. Consequently most of the Nucella used in Littorina—intersex sequence index (ISI ). The reference laboratory examined 133 female Littorina and obtained an this exercise were sub-adults and only a small proportion were the recommended toothed adults. This may have had various overall ISI value of 0.41. Based on these data, the predicted standard error of the ISI on samples of 40 snails is 0.08 ISI consequences for this exercise.Firstly, laboratories may have found it more diYcult to determine the sex of the sub-adults units. As for VDSI in Nucella, the individual laboratory data may be interpreted in terms of Z-scores. The OSPAR guidelines than of toothed adults.Secondly, the sub-adults probably had smaller pene than toothed adults, and consequently any errors state that ISI values should be within 0.5 ISI units of the true value (with 90% confidence). This is approximately equivalent (random or systematic) in the measurement of penis length or observation of reproductive organs would have a potentially to a total allowable error of 0.25 units.On this basis, the data from all laboratories are considered satisfactory (-2<Z<2) greater impact on the final reported values of the summary indices. other than Q156P (questionable, Z=2.36) and Q158P (unsatisfactory, Z=4.76). It is the intention of QUASIMEME to oVer further exercises as part of its overall programme of laboratory performance studies to support marine monitoring activity.Future exercises Littorina—prostate length. The reference laboratory found only one female Littorina with a measurable prostate gland in will take account of the diYculties that were encountered in the current exercise and attempts will be made to obtain more the 133 females that were examined. It is therefore concluded that the population was insuYciently aVected by TBT to allow strongly aVected Littorina. Investigations are required of the possible eVect of transport and storage on the length of the reliable comparisons to be made between laboratories of average female prostate length .All participants, except labora- penis in Nucella. It is also necessary to develop parallel study procedures in relation to the determination of imposex in tory Q154P, agreed with the reference laboratory in that they found 0 or 1 female with a measurable prostate gland.Buccinum undatum.18,19 A further training workshop is planned under QUASIMEME and the EU Biological EVects Quality Assurance in Monitoring Programmes (BEQUALM) pro- Discussion gramme. Particular attention will be paid to accurately sexing specimens, the measurement of penis length and the calculation This exercise has demonstrated that it is possible to conduct international laboratory performance studies for imposex and of summary indices.Discussions are in progress to integrate the laboratory intersex using live marine snails, provided that suYcient care is taken in packaging the animals and that transport time is performance studies of imposex/intersex with the chemical measurements of organotin compounds in shellfish tissue.This kept short. Participants were based in seven European countries and Canada. Where customs authorities did not intervene, combination would reflect the integrated biological–chemical approach to monitoring currently being developed by inter- samples were received in good condition. The success of the study is confirmed by the return of data from 87% of the national monitoring agencies such as OSPAR and the Helsinki Commission (HELCOM).participating laboratories. The data emphasise the care required in preparing sub-samples from a bulk collection of snails. As described Acknowledgements above, most of the samples were prepared by coning and quartering the bulk collection, and these samples showed The authors thank the participating laboratories (Table 5) for submitting their data to the study, the QUASIMEME project relatively small diVerences in mean shell height.Samples distributed on other occasions, for example to replace lost oYce staV (Caroline Warwick, Alison Allen, Kieren Smith, Judith Gourlay) for assistance with the distribution of samples, samples, showed greater diVerences.J. Environ. Monit., 1999, 1, 233–238 2378 B. Bauer, P. Fiorini, I. Ide, S. Liebe, J. Oehlmann, E. Stroben and collation of data and other administrative support, and the B. Watermann, Hydrobiologia, 1995, 309, 15. QUASIMEME scientific group for their support and advice 9 OSPAR, Joint Assessment and Monitoring Programme, Oslo and regarding the exercise.Paris Commission, Guidelines for Contaminant-specific Biological EVects Monitoring, 1997. 10 I. M. Davies, A. Minchin and M. Harding, Marine Laboratory Aberdeen Report 9/97, 1997, p. 33. References 11 D. E. Wells, A. Aminot, J. de Boer, D. Kirkwood and W. P. Cofino, Mar. Pollut. Bull., 1997, 35, 3. 1 G. W. Bryan, P. E. Gibbs and G. R. Burt, J. Mar. Biol. Assoc. 12 W. P. Cofino and D. E. Wells, Mar. Pollut. Bull., 1994, 29, 149. UK, 1988, 68, 733. 13 I. M. Davies, QUASIMEME Bulletin, 1998, Issue 5, pp. 19–20. 2 G. W. Bryan and P. E. Gibbs, ed. M. C. Newman and 14 P. E. Gibbs, P. L. Pascoe and G. W. Bryan, Comp. Biochem. A. W. McIntosh, in Metal Ecotoxicology: Concepts and Physiol., 1991, 100C, 231. Applications, Lewis Publishers, Michigan, 1991, pp. 323–361. 15 P. Fiorini, J. Oehlmann and E. Stroben, Zool. Anz., 1991, 226, 1. 3 P. E. Gibbs, G. W. Bryan, P. L. Pascoe and G. R. Burt, J. Mar. 16 M. Thompson and R. Wood, J. Pure Appl. Chem., 1993, 65, 2123. Biol. Ass. UK, 1987, 67, 507. 17 D. E. Wells and W. P. Cofino, Mar. Pollut. Bull., 1997, 35, 18. 4 S. K. Bailey and I. M. Davies, J. Mar. Biol. Assoc. UK, 1989, 18 C. C. Ten Hallers-Tjabbes, J. F. Kemp and J. P. Boon, Mar. 69, 335. Pollut. Bull., 1994, 28, 311. 5 D. Minchin, J. Oehlmann, C. B. Duggan, E. Stroben and 19 B. P. Mensinck, B. Van Hattum, C. C. Ten Hallers-Tjabbes, J. M. Everaarts, H. Kralt, A. D. Vethaak and J. P. Boon, M. Keatinge, Mar. Pollut. Bull., 1995, 30(10), 633. Nederlands Institut voor Onderzoek der Zee, Rapport, 1997, 6 D. Minchin, E. Stroben, J. Oehlmann, B. Bauer, C. B. Duggan 1997–6. and M. Keatinge, Mar. Pollut. Bull., 1996, 32(2), 188. 7 J. Oehlmann, E. Stroben and P. Fiorini, J. Molluscan Stud., 1991, 57, 375. Paper 9/02797A 238 J. Environ. Monit., 1999, 1, 233–238

ISSN:0960-7919    

DOI:10.1039/a902797a

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

Imposex measurement in the dogwhelk Nucella lapillus (L.)—temporal aspects of specimen preparation† Annagh Minchin*a and Ian M. Daviesa aFRS Marine Laboratory, PO Box 101, Victoria Road, Torry, Aberdeen, AB11 9DB. Fax: +44 1224 295511; E-mail: minchina@marlab.ac.uk; daviesim@marlab.ac.uk Received 9th April 1999, Accepted 30th April 1999 The measurement of the Relative Penis Size Index (RPSI) in the dogwhelk Nucella lapillus (L.) depends upon reliable observations of the length of the penis (PL).Following removal of soft tissue from the shell, PL increases with time leading to a progressively greater over-estimation of the RPSI. The error is proportionally greater at low RPSI values. To retain comparability with previous data, it is suggested that dogwhelks should be examined as soon as possible after shell crushing.for no more than five days. Each shell height was measured Introduction to the nearest 0.1 mm and shells were then crushed and soft The use of tributyltin (TBT) compounds as antifouling agents tissues removed. The sex and VDS of each snail were recorded. since the 1970s led to impacts on non-target organisms.1,2 The PL of each snail was measured at intervals of 45 min Exposure to TBT results in the imposition of male sexual through a working day (6 h, nine observations of each snail ) characteristics (imposex) on female neogastropod snails.3 In to 0.1 mm using a binocular microscope equipped with an severe cases, this can lead to sterility in some species (e.g., the eyepiece graticule.1 These data were converted into millimetres dogwhelk, Nucella lapillus).4 Imposex measurements have been for analysis. Random individual Nucella from the population used widely in surveys of the biological eVects of TBT.4–7 were measured repeatedly over a period of 30 min, up to six Imposex has been adopted by the Oslo and Paris Commission measurements per penis, to account for analytical error.(OSPAR) as a component of their international Joint Assessment and Monitoring Programme (JAMP).The revised OSPAR protocols recommend that snails should be stored live and that the soft tissues should be removed from the shell Results (without narcotisation) prior to examination by microscope.8 The VDSI value for adult dogwhelks from Skatey Shore was Two indices are used to express the degree of imposex; the 3.96 and that for juveniles was 4.0, with an initial RPSI value Vas Deferens Sequence Index (VDSI) and the Relative Penis in adults of 2.5%.The mean shell height of adults was Size Index (RPSI). The Vas Deferens Sequence consists of a 25.48 mm (s 1.45) and of juveniles was 24.98 mm (s 1.60). progression of seven stages from unaVected to sterile females Measurements of PL over 30 min varied by ±0.1 mm and and the VDSI (mean VDS of females) gives an indication of demonstrated a high level of repeatability in relation to both the reproductive competency of the population.The RPSI is the mean PL in the population studied and the change calculated as the cube of the mean length of the penis in induced by delay in the analysis (see below).females (PLF) divided by that in males (PLM), multiplied by a There was a general tendency for PL of both male (PLM) hundred. Reliable measurements of PL are therefore important and female (PLF) Nucella to increase with time after removal as they have direct impact on the calculated RPSI values. of the animals from their shells. The average PLM increased It is common laboratory practice to prepare batches of from 2.71 (s 0.44) to 3.19 (s 0.44) mm and PLF from 0.79 (s snails (e.g., by sampling location) for examination and some 0.31) to 1.35 (s 0.29) mm after 6 h.The average PLM of juvenile time may therefore elapse between removal from the shell and males increased from 1.55 (s 0.39) to 1.93 (s 0.49) mm and measurement of the PL. This may result in variation in penis that of juvenile females from 0.69 (s 0.20) to 0.99 (s 0.23) mm length among individual Nucella, due to the time elapsed from after the same period.The increases mainly took place during the removal of soft tissue from the shell. As continuation of the first three hours, with relatively small changes between an investigation of sample preparation/conservation,9 this four and 6 h (Fig. 1). The increase in PL was proportionally paper examines the changes in PL with time in Nucella which much greater (Fig. 2) in animals with a small penis (e.g., adult have been removed from their shells and assesses the females showed an average increase by a factor of 1.7) than significance of these changes for RPSI determination. in those with a relatively large penis (e.g., adult males, factor of 1.2).Juveniles tended to show less of an increase in PL Methods than did adults of similar initial PL (Fig. 3). Linear regressions were fitted to the relationships between N. lapillus were collected from below the mid-shore level at the initial penis length (PL0) and the penis length after 6 h Skatey Shore, Stonehaven, east Scotland on 30 January (100 (PL6) for both adult (Fig. 4) and juvenile dogwhelks: juveniles) and 4 June (200 adults) 1998. They were maintained at ambient temperature in flowing sea water at the laboratory Juveniles: PL6=1.052 PL0+0.270 (R2=0.9430, n=99) †©Crown copyright. Adults: PL6=0.948 PL0+0.611 (R2=0.9640, n=200) J. Environ. Monit., 1999, 1, 239–241 239Fig. 1 Changes in the ratio of mean penis length at time t (PL-t) to Fig. 4 Relationship between the ratio of RPSI values calculated after initial mean penis length (PL-0) over a period of 6 h for adult and 6 h (RPSI6) to the initial RPSI value (RPSI0) and the initial RPSI juvenile Nucella lapillus. value for adult Nucella lapillus. of imposex in adults (RPSI 10–50%), RPSI would be increased to 17–57% (i.e., factors of 1.7–1.14), but at low RPSI values (<2%) the proportional changes would be very much greater.Huet10 studied the eVect of narcotisation of Nucella by MgCl2 solution before examination. Several field studies have utilised this procedure.3,10,11 Narcotisation led to an extension of the penis that was proportionally greater at high penis lengths. The RPSI of narcotised Nucella was approximately 80% of that of non-narcotised snails over a wide range of RPSI values, and Huet10 suggested that a conversion factor of 0.8 could be used.While narcotisation leads to under estimation of the RPSI, delay in the measurement of untreated snails leads to over-estimation. Fig. 2 Relationship between the ratio of penis length after 6 h (PL-6) The observations in this report suggest that the most to initial penis length (PL-0) and initial penis length for adult and juvenile Nucella lapillus.reproducible results would probably be obtained from snails which had been left out of the shell for a period in excess of four hours. However, most published data have been obtained Discussion from fresh material, as recommended in the OSPAR guidelines Imposex analysis, generally, only requires one PL measurement and adopted in recent international laboratory performance and not repeated measurements of the same individual over studies under QUASIMEME.12,13 In order to retain compartime.The time between removal from the shell and the ability with previous data, it is advisable to examine dogwhelks measurement of the PL will aVect RPSI calculations as, once as soon as possible after extraction from the shell.It is not removed from their shells, PL in snails increased. Juvenile and recommended that PL be ‘corrected’ to some standard elapsed adult females show more of an increase than males. This time by the application of regression equations, such as those resulted in changes in RPSI from initial values of 2.5% in developed above.adults and 9.0% in juveniles, to 7.5% and 13.7%, respectively, over the 6 h period. The corresponding changes in Relative Conclusion Penis Length Index (RPLI) were from 29.3% and 44.8% to 42.2% and 51.5% over the same interval and probably reflect Penis lengths, and hence the RPSI values, of Nucella increase gradual relaxation and senescence of the snails. with time after removal from the shell.Penis lengths stabilise The regression equation above can be used to estimate the after three to four hours, but to retain comparability with potential eVect of this aspect of sample handling on popu- previous data, it is recommended that all snails be measured lations showing a range of RPSI values. At intermediate levels as soon as possible after removal from the shell.Acknowledgements We would like to thank Matt Gubbins, Patricia Barrett and Gavin Grewar for their help with the collection and processing of samples. References 1 G. W. Bryan, P. E. Gibbs, L. G. Hummerstone and G. R. Burt, J. Mar. Biol. Assoc. UK, 1988, 66, 611. 2 E. Stroben, J. Oehlmann, U. Schulte-Oehlmann and P. Fioroni, Malacol. Rev. (1996), Suppl. 6 (Molluscan Reprod.), 1996, 173. 3 J. Oehlmann, E. Stroben and P. Fioroni, J. Molluscan Stud., 1991, 57, 375. Fig. 3 Correlation between penis length measured after 6 h (PL-6) and 4 P. E. Gibbs, G. W. Bryan, P. L. Pascoe and G. R. Burt, J. Mar. Biol. Assoc. UK, 1987, 67 507. initial penis length, for adult and juvenile Nucella lapillus. 240 J. Environ. Monit., 1999, 1, 239–2415 G. W. Bryan and P. E. Gibbs, in Metal Ecotoxicology: Concepts 10 M. Huet, P. Fioroni, J. Oehlmann and E. Stroben, Hydrobiol., 1995, 309, 29. and Applications, ed. M. C. Newman and A. W. McInthosh, Lewis, Ann Arbor, 1991, pp. 323–361. 11 A. Minchin and D. Minchin, Environ. Technol., 1997, 18, 1225. 12 I. M. Davies, QUASIMEME Bulletin, 1998, Issue 5, pp. 19–20. 6 D. Minchin, J. Oehlmann, C. B. Duggan, E. Stroben and M. Keatinge, Mar. Pollut. Bull., 1995, 30(10), 633. 13 QUASIMEME, Report of exercise 359, BE1 Fisheries Research Services, Marine Laboratory, Aberdeen, 1998, 7 pp. 7 J. Svavarsson and H. Skarphe�dinsdo� ttir, Sarsia, 80(1), 35. 8 I. M. Davies, A. Minchin and M. Harding, Marine Laboratory, Aberdeen Report No 9/97, 1997, 33 pp. Paper 9/02836F 9 A. Minchin and I. M. Davies, J. Environ. Monit., 1999, 1(2), 203. J. Environ. Monit., 1999, 1, 239–241

ISSN:0960-7919    

DOI:10.1039/a902836f

出版商:RSC    

年代:1999

数据来源: RSC

摘要:

Establishment of a sampling strategy for the use of blue mussels as an indicator of organotin contamination in the coastal environment Koji Shindo and Akira Otsuki* Department of Marine Science and Technology, Tokyo University of Fisheries, 4-5-7 Konan, Minato-Ku, Tokyo 108, Japan. E-mail: md97012@edufs.tokyo-u-fish.ac.jp (Koji Shindo); akirao@s4201.tokyo-u-fish.ac.jp (Akira Otsuki); Fax:+81 3 5463 0398; Tel:+81 3 5463 0458 Received 10th December 1998, Accepted 16th March 1999 The application of the Mussel Watch concept to the assessment of chemical contamination in the coastal environment is still premature, since the relationship between the physiological and ecological aspects of blue mussels and the accumulation of contaminants in their soft tissues remains unclear. We cannot yet directly estimate with known confidence the degree of chemical contamination from the levels of contaminants in the soft tissues of mussels. An understanding of the source and range of variability in the tissue concentrations of contaminants is essential, and the establishment of a biomonitoring sampling strategy to minimize the eVect of identified sources of variability is required.The present study was conducted to clarify the characteristics of organotin accumulation in blue mussels under various conditions in Tokyo Bay, and to establish an optimized sampling strategy of mussels as exposure indicators of organotin contamination in Japan. It was clear that the sample number, individual size, spawning activity and vertical habitat were factors causing a variation of tissue concentration.Based on a quantitative estimation of the variability of organotin concentration in mussel tissues under various physiological and natural conditions, we suggest that a composite sample of 30 mussels (3–5 cm in shell length), collected from the infralittoral zone prior to their spawning season, is essential to reduce the variability between individuals and to obtain reproducible analytical values at a sampling site.For monitoring sites where natural blue mussels cannot be collected, an alternative method of transplanting blue mussels from a relatively clean area would be required. accumulation in mussel tissue. Little research on the physio- Introduction logical and ecological factors aVecting the bioaccumulation of The Mussel Watch concept, based on the bioaccumulation of contaminants in blue mussels has been performed.As a contaminants by natural blue mussels, is becoming a worldwide consequence, we cannot yet completely validate the origin and monitoring system of chemical contamination in the coastal range of variability of contaminant concentration in biological environment, because the analysis of contaminants in mussel tissues, or the spatial or temporal coverage of contamination tissues is regarded as a superior way of providing information expressed by only one value of tissue concentration.The about the occurrence and distribution of contaminants.1,2 answer to the origin and range of variability of tissue concen- Nowadays, many studies are providing us with a rough tration is directly connected to the establishment of a sampling approximation of the status and trends of contaminants in the strategy that will minimize the eVect of identified sources of coastal environment.3–6 The genus Mytilus consists of a group variability. An optimized sampling strategy should provide us of widespread sedentary mussels.Many contaminants accumu- with a method of collecting a ‘reliable sample’ that reflects the late in their soft tissues to high concentrations, making the contaminant levels in ambient seawater for the assessment of chemical contamination.Thus, such a strategy is also essential measurement of such chemicals easier than in seawater for clarifying the limits of its applicability as a biomonitor- samples.The tissue concentration of contaminants can be ing method. regarded as a time-averaged or time-integrated concentration The purposes of this study were to clarify the characteristics for the ambient environment. This implies that the Mussel of organotin accumulation by blue mussels under various Watch programme may become a useful coastal monitoring conditions in Tokyo Bay, Japan, and to establish an optimized method.However, the application of biomonitoring using blue sampling strategy of mussels as an indicator of exposure to mussels to assess chemical contamination has several important organotin compounds in Japan. The optimized sampling strat- problems in spite of careful sampling and chemical analysis: egy and its applicability as a monitoring method for six (i) taxonomic diYculties in sampling; (ii) lack of prescribed organotins were considered in terms of the usefulness of this pertinent sampling/analytical strategies; (iii) uncertainties in technique as an analytical method for the determination of variability and spatial or temporal resolution of tissue concenorganotin compounds in mussel tissue and with regard to the tration; and (iv) limitations of the approach itself.7,8 The most variability of tissue concentration due to the ecological aspects popular strategies of the National Oceanic and Atmospheric of blue mussels.Administration (NOAA) in the USA and the Re�seau National d’Observation (RNO) in France are no exception to these Materials and methods problems. The most basic issue in these problems seems to be a lack of understanding of how important these factors are in Sample collection and preparation the production of reliable data, although it is well known that a variety of factors influence the relationship between the Natural blue mussels were collected by hand from three stations in Tokyo Bay, located in the central part of Japan concentration of contaminants in ambient water and the J.Environ. Monit., 1999, 1, 243–250 243(Fig. 1): Tokyo-Light, located oVshore, is an artificial construction supported on a steel pole; Odaiba is an artificial rocky reef in the recesses of the Bay; Kannonsaki is a natural rocky reef located in the mouth of the Bay. Blue mussels were sampled mainly in June, August, November, February and April from 1994 to 1996, based on their ecological characteristics in Tokyo Bay (Fig. 3, see later). The identification of blue mussels was based on the characteristics of the shell shape and colour of the mantle edge and shell.9 Mussels were collected as a composite sample of individuals from several single clumps within a few square feet at the lower tidal level (±0 m). We selected 20–150 mussels per analytical sample according to the objectives.The samples were kept in cold storage (<10 °C) until pretreatment in the laboratory. Pretreatment was carried out within the day of sampling. Water retained inside the shell was completely removed with a stainless strainer, the byssus was discarded and the mean whole wet weight of the sampled mussels was measured.Mussels from each sampling site and date were divided into six size groups according to shell length: 1.5–1.8 cm, 1.8–3 cm, 3–4 cm, 4–5 cm, 5–6 cm and 6–7 cm. Fig. 2 The equipment for transplanted bivalves. After taking out the soft tissue, the adhering water was fully removed by draining for 20 min in a stainless strainer. For the study on organotin distribution in the soft tissues, 50–70 individuals (3–5 cm in shell length) were dissected into gill, ation and gas chromatography with flame photometric viscera, mantle and muscle.Finally, each sample was separately detection. Details of the analytical procedure are given in homogenized (10 000 rpm, 10 min) and then frozen (-40 °C) previous papers.12–16 until analysis. Details of sample handling, pretreatment and Each organotin peak in the environmental samples was storage followed the previous papers.10,11 verified by the addition of organotin standards.Based on the internal standard method, the concentrations of the six ana- Transplantation procedure lytes in the extract were calculated by comparing the peak area of the spiked internal standard [tripentyltin chloride Natural blue mussels for transplant were collecteby hand (TPeT)] with that of each analyte.Organotin values in the from a relatively uncontaminated site in Tokyo Bay (Iwai or extract were expressed as the chlorides. The limit of detection Tateyama) or a seashore rocky reef facing the Pacific Ocean (S/N=2) was 0.5–0.7 ng (chloride) g-1 wet weight for the six (Kujukuri) outside Tokyo Bay.The mussels were kept in organotins in mussel samples. The reported concentration was uncontaminated seawater for about 1 month before transplanusually expressed as a mean value of three determinations tation. They were transplanted to Tokyo-Light and Hukuura from one extract, and was corrected by the reagent blank and Harbour. We transplanted 30 individuals (3–5 cm in shell recovery of each organotin species.length) into each of five positions with respect to tidal height for about 1 month in autumn and winter from 1994 to 1997 (Fig. 2). Pretreatment of tissue samples was performed in the Results and discussion same way as for natural mussels. Evaluation of the analytical method for the determination of organotin compounds in blue mussel soft tissue Analytical procedure Recovery was validated by comparing the results of six repli- The determination of six organotin compounds [tributyltin cate analyses of an uncontaminated sample spiked with (TBT) and triphenyltin (TPT) and their degradation products] 16–28 ng (chloride) g-1 wet weight (this value was the average in homogenized soft tissue (3 g) of blue mussels was carried level of samples from Tokyo Bay) with those obtained from out using sonication with tropolone–benzene, propyl derivatizthe analysis of propylated standards.Table 1 indicates the recovery rate of 10 organotin chlorides in blue mussel tissue through the whole analytical procedure. The recovery by our procedure was in the range 56–116% for butyltin and phenyltin compounds. Repeatability was assessed with five replicate analyses of three average contaminated samples.The relative standard deviation (RSD) ranged from 9.4 to 26% for the 10 analytes and was 8.2% for TPeT. The accuracy of our analytical procedure was validated by five replicate analyses of 2 g of certified reference material from the National Institute for Environmental Studies (NIES No. 11 Fish Tissue).Our method yielded 69.9±7.2% of the TBT certified value and 50.4±8.6% of the TPT reference value. It should be noted that the physicochemical form of the analyte in the spiked sample is probably diVerent from that in the real sample, especially in the analysis of biological tissues where the organotin compounds may be bonded to lipids or proteins, and that the use of spiked samples can lead to an overestimation of the extraction eYciency.17 Thus, the Fig. 1 Sampling stations in Tokyo Bay. values of the recovery for spiked samples may be higher than 244 J. Environ. Monit., 1999, 1, 243–250Table 1 The recovery of organotin compounds in bivalve samples by our analytical procedure MMT DMT TMT MBT DBT TBT MPT DPT TPT Recovery (%) 135.7 80.4 46.2 115.9 100.5 81.0 56.0 111.8 95.5 RSD (%) 13.1 13.8 25.6 20.7 18.9 14.8 26.1 9.9 13.2 Number of analyses n=6.TPeT: absolute recovery, 79.0%; RSD, 18.2%. MMT, monomethyltin; DMT, dimethyltin; TMT, trimethyltin; MBT, monobutyltin; DBT, dibutyltin; TBT, tributyltin; MPT, monophenyltin; DPT, diphenyltin; TPT, triphenyltin. those in real soft tissues, and our analytical results might be temporal bias [Table 2(a)].In spring, blue mussels less than 1.8 cm in shell length were few and, in summer, many mussels 30–50% lower than the concentrations in real samples. Judging from the recovery of spiked samples using a highly longer than 5 cm disappeared. This seasonal population change suggests that mussels less than 1.8 cm in shell length reproducible analytical method, our analytical procedure is a reliable method for organotin analysis in mussel soft tissues, and those longer than 5 cm may not always be sampled, while blue mussels of 3–5 cm in shell length can be collected although the results obtained from the recovery test may not completely reflect the amount of organotin compounds throughout the year.Thus, the number and size of accessible mussels on each sampling date may be greatly limited by extracted from real tissue samples.The analytical variability was evaluated to be <5–26% RSD of the tissue concentration. their population fluctuation at each sampling site. Table 2(b) shows the seasonal soft tissue weight of individuals in our sampling stations; the seasonal variation was within the range Sampling of natural blue mussels 2–20%.Our results based on the population dynamics and Although mussels are dominant organisms in the intertidal average weights of individuals of naturally occurring blue sessile fauna in Tokyo Bay, taxonomic identification of Mytilus mussels suggested that individuals of 3–5 cm in shell length during sample handling enabled us to discriminate easily between blue mussels and other mussels and bivalve genera.Blue mussel samples could easily be collected from each sampling site. However, we cannot yet discriminate between Table 2 Population dynamics and sampling availability of natural two blue mussel species (M. galloprovincialis and M. edulis), mussels at sampling stations using morphological characters, allozyme characters and reproductive patterns.Blue mussels in Tokyo Bay were tenta- (a) tively identified as Mytilus edulis galloprovincialis.18 Fig. 3 Sampling availability of natural mussels shows the relationship between the ecological characteristics and sampling time of blue mussels in Tokyo Bay. Their Individual size/cm ecological characteristics indicate that their physiological condition and population size change drastically within a year, as Sampling date 1.0–1.5 1.5–1.8 1.8–3 3–4 4–5 5–6 6–7 7–8 a result of the intense heat on summer days, spawning activity June ’94 + + ++ ++ ++ + - N and recruitment of young mussels.18–20 Aug.’94 + + + +++ N N N The spatial and temporal distribution, individual size and Nov. ’94 - ++ ++ + + N N N volume of soft tissue of blue mussels under various natural Jan.’95 - ++ ++ + + - N N conditions were examined as primary factors in sample Feb. ’95 - + ++ ++ + - N N handling, because the Mussel Watch concept depends on the Apr. ’95 ++ + ++ ++ ++ - - N presence of natural mussels. Our observation of the shore June ’95 + ++ ++ +++ + -N Aug. ’95 + ++ ++ +++ - -N surrounding Tokyo Bay indicated that the distribution of Jan. ’96 - - + + - - N N blue mussels was patchy, although their population was large Feb.’96 N N + + - - N N in each habitat. This means that we cannot necessarily collect May ’96 + + + ++++- -N blue mussel samples in locations where we need to monitor N, none present or no sampling; -, few present (need for hard contaminants. The seasonal size composition of blue mussels sampling); +, enough present for sampling; ++, many present.in the lower tidal level of our sampling stations indicated that the presence of small and large individuals had a (b) Ratio of average weight to the value of 4–5 cm natural mussel samples Individual size/cm Sampling date 1.5–1.8 1.8–3 3–4 4–5 5–6 6–7 June ’94 0.07 0.22 0.50 1 (9.5)a 1.6 2.3 Aug. ’94 0.10 0.22 0.76 1 (6.9)a N N Nov. ’94 0.07 0.21 0.56 1 (9.3)a N N Jan.’95 0.09 0.13 0.62 1 (9.1)a 1.4 N Feb. ’95 0.07 0.19 0.61 1 (9.9)a 1.3 N Apr. ’95 0.03 0.23 0.64 1 (8.8)a 1.5 2.4 June ’95 0.05 0.20 0.54 1 (9.1)a 1.6 2.3 Aug. ’95 0.07 0.18 0.60 1 (8.2)a 1.2 1.9 Jan. ’96 0.05 0.11 0.42 1 (3.6)a 1.8 N Feb. ’96 N 0.18 0.50 1 (2.8)a 2.0 N May ’96 0.02 0.18 0.55 1 (3.5)a 1.6 3.2 aThe mean of actual wet weight (g wet). N, none present or Fig. 3 Ecological characteristics and sampling time of natural blue no sampling. mussels in Tokyo Bay. J. Environ. Monit., 1999, 1, 243–250 245were the most average-sized adults growing steadily in rapidly and intestinal tract from ambient water and food. A strong aYnity of organotin compounds with mucin in gills has also changing populations. Judging from the average wet weight of individuals, the been reported.22 It is suggested that organotin compounds may primarily accumulate in gills and viscera, and be second- minimum number of individuals required for repeatable analysis of the six organotins was a composite sample of more than arily transferred into lipid- and protein-rich tissues, mantle (including gonadal tissue) and muscle by circulatory transport.five mussels of 5–6 cm in shell length, more than 12 mussels of 3–4 cm in length or more than 150 mussels smaller than The four diVerent tissues showed almost the same accumulation patterns of TBT, TPT and their degradation products 1.8 cm in length (Table 3). However, BeliaeV et al.3 and Daskalakis et al.21 stated that a suYcient number of individuals in all seasons.This similarity suggests that blue mussels do not metabolize the tri-form of organotin compounds to di- to average the residual diVerences between individuals was a composite sample of more than 30 bivalves, and that the use and mono-forms in soft tissues. This suggestion is in agreement with the finding that the detoxifying activity of mussel tissue of more than 30 individuals from each population is suYcient to permit eVective comparison (detect diVerences of 40%) of for organotin compounds is very low.23,24 A significant decrease in the level of accumulation of the tissue concentrations in these two instances.Therefore, on the assumption that more than 30 individuals of the same shell six organotins in mantle tissue during the spawning period was observed, although the accumulation pattern and level of length collected from a single clump within a few square feet in the infralittoral zone is enough to average the residual the six compounds in the four diVerent tissues did not indicate a definite seasonal variation.The decrease in accumulation diVerences between individuals, and that all collected individuals been exposed to a similar environmental and physiological level showed the same tendency as the change in the gonad index at spawning time.Previous reports25–27 demonstrated conditions, we focused our discussion on two factors: (i) the characteristics of organotin accumulation in blue mussels; and that many contaminants are incorporated into mussel gonad or bonded to lipids and proteins in gametes, and that fluctu- (ii) the establishment of an optimized sampling strategy.Moreover, we individually estimated the variability associated ations in the accumulation of contaminants were correlated with the seasonal cycle of gametogenesis and spawning activity. with sampling date, sex diVerence, individual size and habitat height with application of the optimized sampling strategy for This implies that organotin compounds accumulated in the gonad (or mantle) are released with eggs and sperm during three parameters except the investigated parameter.The weight of variability due to each investigated parameter was statisti- spawning (Fig. 4). Organotin compounds released with gametes were estimated at 12–63% of their accumulated con- cally calculated by relative comparison between the analysed values of the six organotins in an optimized mussel sample centration in mantle tissue.Errors due to sampling date, especially in the spawning period, were estimated to be and in mussels under the influence of the investigated ecological parameter. 2.5–18.4% of the analysed values derived from a reliable mussel sample prior to the spawning season. Accumulated concentration in each organ under various physiological conditions DiVerences in accumulated organotin concentration due to sex, shell length and vertical habitat The influence of the reproductive and other seasonal cycles on the accumulated concentration in each organ of the mussels Sexual diVerence in accumulated concentration.On each sampling date, we collected individuals whose sex was discern- was examined by determining the accumulated organotin concentration in 50–70 individuals of 3–5 cm in shell length.ible by the colour of the mantle. We determined the accumulated organotin concentration in 50–70 (male or female) First, the physical condition and reproductive activity of the mussels were observed under natural conditions.In summer individuals of 3–5 cm in shell length. Mantles of female and male mussels showed almost the same accumulation pattern and just after the spawning season, soft tissues obviously lost total weight [3–14%; Table 2(b)]. The composition of the of TBT, TPT and their degradation products under seasonal and reproductive cycles (Fig. 4). Variability due to sex diVer- mantle including gonadal tissue showed a seasonal variation (minimum, 9%; maximum, 30%).The gonadal index calculated ence was estimated to be 1.4–7.8%. With regard to the accumulation of organotin compounds in soft tissue in blue in blue mussels of 3–5 cm in shell length showed the same tendency as that observed previously.18 Almost all mature mussels, it is clear that sex is not an important variation factor.individuals seemed to have finished releasing gametes by April. However, we could diVerentiate the sex of some individuals in Organotin accumulation levels in mussels of diVerent size. Seasonal accumulation levels for diVerent shell lengths under the population by the colour of the mantle not only from mid- October to March, but also in summer. various physiological conditions were examined by determining the accumulated organotin concentrations in 25–150 individ- The accumulated concentration of organotin compounds in each tissue increased in the order of gill, viscera, mantle and uals of each shell length.All mussels had higher levels of organotin compounds in their tissue than those present in muscle; gill and viscera concentrations were 1.2–4 times higher than those of other tissues, showing that the mussels accumu- ambient seawater.However, no relationship was found between accumulation level in whole soft tissues and shell late high concentrations of the six organotins in gill, viscera Table 3 Sample collection of natural blue mussels Shell length/cm 1.5–1.8 1.8–3 3–4 4–5 5–6 6–7 3–5a Average weight of 0.48–0.72 1.0–2.1 2.7–6.4 6.1–9.2 9.4–16 15–22 4.6–9.2 individuals/g wet Minimum number for 150 30 12 10 5 3 70 repeatable analysis Number of collected mussels 150 100 50 30 25 25 100 aSample of various organs. 246 J. Environ. Monit., 1999, 1, 243–250Fig. 4 Organotin concentration in mantle including gonadal cell and gonad index of natural mussels. length (Table 4). Accumulation levels in mussels of diVerent Accumulated organotin concentration in diVerent vertical habitats.The diVerence in vertical position of mussel habitats shell lengths did not show significant seasonal variation. Small mussels, 1.5–1.8 cm in shell length, often had 1.1–3 times relates to the immersion time in seawater. Judging from the average condition of Tokyo Bay and the tidal level data higher levels of organotin compounds than those in mussels of 3 cm or above.Mussels of 3–5 cm in shell length (1–2 years calculated by the harmonic analysis of tides, the submerged ratio of mussels in the infralittoral zone is 0.5–10% of sub- old) showed nearly the same concentration level and accumulation patterns of organotin compounds in whole soft tissue merged time in the intidal zone (Fig. 6). The drying of mussels when out of seawater negatively aVects their growth rate and on every sampling date. Compared with large adult mussels longer than 3 cm in shell reproductive activity. The growth of mussels in our samples before and after transplantation showed the same tendency as length, mussels of less than 1.8 cm being recruited to mussel beds in each spring show rapid feeding and a high growth in the previous reports.18,19 Compared with adult (3–5 cm) mussels before transplantation, those transplanted to the tidal rate, but a low excretion rate of extraneous substances.19,28,29 An absence of correlation between shell length and lipid zone (above the mid-tide level ) showed some mortality and 60–110% growth in wet weight of the survivors; those trans- content has also been reported.30 The biological characteristics of small mussels ( less than 1.8 cm long and less than 1 year planted to the infralittoral zone (beneath the mid-tide level ) showed 70–140% growth in wet weight (Fig. 6). Transplanted old) would lead to a high accumulation of contaminants. Moreover, the eVect of reproductive activity on the accumu- mussels showed little increase in shell length after transplantation.lation of organotin compounds in soft tissue is significant before and after spawning time, because there is a large We determined the accumulated organotin concentrations in 30 individuals of 3–5 cm in shell length transplanted to five diVerence in accumulation characteristics between mature adult ( longer than 3 cm) and immature ( less than 1.8 cm long) habitats at diVerent tidal heights.The results showed that the habitat position with respect to the tide caused significant mussels. The variability due to individual size was estimated to be ±6–38% for TBT and 10–116% for TPT of the analysed diVerences in accumulated organotin concentrations. Mussels in the infralittoral zone (from average lower tidal level to value derived from 3–5 cm mussels.The population dynamics and average weight of individuals [Tables 2(a) and 2(b)] sublittoral zone) were 1.2–1.5 times stronger accumulators than those in the intidal zone (Table 5). However, the mussels suggested that the 3–5 cm mussels were the most ‘average’ adults. Mallet and Carver31 and Lowe et al.32 reported that in these five diVerent tidal levels showed nearly the same accumulation patterns of the six organotin compounds in the normal 1–2-year-old mussels have a stable internal body condition for growth and reproduction under rapidly changing whole soft tissue under various seasonal conditions.The eVect of an increase in shell length on the accumulation level can be natural conditions, and indicated the similarity in reproduction and internal body condition between such blue mussels.Our disregarded, because our long-term field study made it clear that the uptake of organotins in the soft tissue of transplanted results suggest that the similarity of level and pattern of organotin accumulation in mussels of 3–5 cm in shell length blue mussels was rapid and that the apparent equilibrium of organotin concentration between the soft tissue and ambient in our sampling sites is dependent on the similarity in reproduction activity and internal body condition of the mussels seawater required only 2–3 weeks.33 The diVerence in height of the habitats resulted in diVerent immersion (or exposure) (Fig. 5). Judging from sample handling and the similarity of accumulation in tissue, more than 30 mussels of 3–5 cm in times in contaminated seawater.The variability due to habitat height was estimated to be ±10–27% for TBT and 2–64% for shell length are suitable as a composite sample for use as an exposure indicator. TPT of the average value for mussels from the lower tidal level. This variability suggested that the immersion time in seawater aVects not only the advantage of habitat but also the Table 4 Variability of organotin concentration in diVerent sized accumulation level of organotins.natural mussels in Tokyo Bay Optimized sampling strategy Ratio of concentration to the value of 3–5 cm sample Currently, the most popular programmes, those of the NOAA34 and RNO,6 have a regularized method to collect 1.5–1.8 cm 1.8–3 cm 3–5 cm 5–6 cm 6–7 cm ‘reliable samples’. NOAA’s sampling strategy attempts to limit environmental influences by collecting annually a set number TBT 1.38 1.23 1 (19.3)a 0.84 0.89 (30 individuals ×3) of blue mussels with a set size range TPT 1.13 1.24 1 (4.48)a 0.90 2.16 (5–8 cm) at the same time (in winter prior to the spawning aThe mean of actual concentration value (ng g-1).season), at the same depth (continuously submerged mussels) J.Environ. Monit., 1999, 1, 243–250 247Fig. 5 Seasonal variation of organotin accumulation in mussels of 3–5 cm in shell length. Fig. 6 Actual submerged ratio of mussels in five vertical habitat positions and the growth of mussel samples before and after transplantation. Table 5 Variability of organotin concentration in mussels transplanted to five diVerent tidal heights in Tokyo Bay Ratio of concentration to the value of sample in average tidal level Upper limit of Middle of Average of lower Sublittoral Lower limit of distribution tidal level tidal level zone distribution TBT 0.896 0.732 1 (72.4)a 0.939 0.827 TPT 1.02 1.45 1 (6.58)a 1.64 1.21 aThe mean of actual concentration value (ng g-1).and in the same site every year.The purpose of the composite collect native blue mussel samples in locations where measurements are required to monitor contaminants; (ii) population samples in one sampling site is to minimize sample variability. To reduce variability due to spawning state and hydrography, fluctuations within a sampling site greatly limit the number and size of accessible mussels on each sampling date; (iii) the all sampling has to be made within±3 weeks of a certain date every year.RNO’s sampling strategy attempts to take within- reproductive cycle aVects the accumulated concentration, especially due to the significant decrease in accumulated year fluctuations in chemical concentrations into account by collecting seasonally a set number (50 individuals, a number organotins in mantle tissue during the spawning season, while blue mussels show nearly the same accumulation pattern in high enough to minimize the eVect of individual variations) of blue mussels with a set size range (3.5–6 cm) at the same site all seasons; and (iv) diVerences in immersion time in seawater resulting from habitat height generate diVerences in exposure (intertidal zone) in autumn every year.However, these regulations only indicate sample number, size, sampling location, time to organotins. Based on the eVect of these ecological aspects when mussels are used for monitoring organotin date of sampling and the reasons to regularize sampling. Therefore, we clarified the characteristics of organotin accumu- contamination, we propose a sampling strategy for blue mussels as exposure indicators in Japan as follows.lation in terms of the ecological aspects of mussels under various natural conditions. (i) Sampling location: a large number of individuals must be collected from several clumps within a few square feet at Our quantitative estimates of the variability of organotin concentrations in diVerent mussel samples under natural con- the same depth (continuously submerged mussels), such that all mussels essentially receive similar environmental stress and ditions suggest the eVect of ecological aspects when blue mussels are used as exposure indicators: (i) we cannot always exposure conditions. 248 J. Environ. Monit., 1999, 1, 243–250(ii) Sampling date: to reduce variability due to spawning, mussels not only restricts the number and size of accessible samples, but also prevents sampling in certain locations where samples must be collected from the same sampling locations at the same time, prior to the spawning season. contaminants should be monitored; (ii) we cannot yet directly estimate with known confidence the degree of chemical con- (iii) Sample number: a composite sample of more than 30 individuals from each population is required for satisfactory tamination based on the levels of accumulation of contaminants in the soft tissues of mussels.Further studies based on reduction of sampling variance. (iv) Size of mussels: collected mussels should have the an optimized sampling strategy should reveal with known confidence the temporal and spatial coverage of ambient narrowest possible range of shell length in order to reduce satisfactorily ecological and physiological diVerences between contamination drawn from mussel tissue concentrations, and the capabilities and limitations of the Mussel Watch concept individuals (an average length of 3–4 cm or 4–5 cm presents least variability).as a monitoring method for chemical contamination.(v) Availability of samples: some requests for contaminant monitoring may be in locations where no natural mussels are Acknowledgements living; we must standardize a transplantation method as an alternative for such cases. We thank the staV of Tokyo-Light, Banda Field Marine Advances in analytical instrumentation enable us to analyse Laboratory and the National Institute of Fisheries Science for trace levels of many contaminants in biological tissues.We their support. We also give special thanks to Drs. Hiroshi can solve analytical problems by the spread of standardized Yamakawa and Osamu Oku for their advice. analytical procedures and certified reference mussel tissue material. Thus, it is increasingly recognized that the major risks of error in environmental monitoring are not in the References laboratory (analysis of contaminants) but rather during field 1 M.E. Gurtz, in Biological Monitoring of Aquatic Systems, ed. S. L. operations (sampling) or prior to analysis (sampling, storage Loeb and A. Spacie, Lewis, Florida, 1995, pp. 345–347. and sample pretreatment).10 Pretreatment of mussel samples 2 E.Goldberg, Environ. Monit. Assess., 1986, 7, 91. after collection produced another suggestion for the optimized 3 B. BeliaeV, T. P. O’Connor, D. K. Daskalakis and P. J. Smith, sampling strategy. The analysis of only one organ of blue Environ. Sci. Technol., 1997, 31, 1411. 4 G. G. Lauenstein, Mar. Pollut. Bull., 1995, 30, 826. mussels as an exposure indicator should be avoided, because 5 M.Morita and Y. Shibata, Japanese–French Workshop on Recent the analysis of contaminants in each organ requires not only Progress on Knowledge of the Behavior of Contamination in large individuals and extra treatments for the organs involved, Sediments and Their Toxicity to Aquatic Organisms, Workshop but also additional information about contaminants in an Proceedings, National Institute of Fisheries Science, Yokohama, organ.For example, it has been reported that contaminants 1994, pp. 142–154. included in the undigested remains in the intestinal tract of 6 E. I. Hamilton, Mar. Pollut. Bull., 1989, 20, 523. 7 D. J. H. Phillips and D. A. Segar, Mar. Pollut. Bull., 1986, 17, 10. mussels can clearly overestimate the accumulated concen- 8 D. Cossa, Oceanol.Acta, 1989, 12, 417. tration in viscera.35 However, we cannot yet estimate the 9 E. Gosling, in The Mussel Mytilus: Ecology, Physiology, Genetics degree of error caused by imprudent pretreatment of undiand Culture, ed. E. Gosling, Elsevier, Amsterdam, 1992, pp. 3–17. gested remains in the intestinal tract. It can be concluded from 10 K. J. M. Kramer, in Quality Assurance in Environmental these results that a composite sample of whole soft tissue from Monitoring—Sampling and Sample Pretreatment, ed.Ph. 30 blue mussels of 3–5 cm in shell length, collected from the Quevauviller, VCH Press, Berlin, 1995, pp. 179–211. 11 A. M. Caricchia, S. Chiavarini, C. Cremisini, R. Morabito and R. infralittoral zone at a time prior to the spawning season, is Scerbo, Anal. Chim.Acta, 1994, 286, 329. essential to reduce the variability between individuals under 12 J. L. Gomez-Ariza, E. Morales, R. Beltran, I. Giraldez and M. natural conditions and to obtain reproducible analytical values Ruiz-Benitez, Analyst, 1995, 120, 1171. at each sampling site. 13 S. Ohhira and H. Matsui, J. Chromatogr. B, 1990, 525, 105. The present study also revealed some variability due to the 14 H.Hraino and M. Fukushima, Anal. Chim. Acta, 1992, 264, 91. ecological aspects of mussels when used as exposure indicators 15 J. A. Sta�b and U. A. Th. Brinkman, Appl. Organomet. Chem., 1994, 8, 577. for organotin contamination. The multiple weight of the 16 Y. K. Chau, F. Yang and M. Brown, Anal. Chim. Acta, 1997, variability associated with the sampling date, sex diVerence, 338, 51.individual size and habitat height was estimated using a 17 M. Abalos, J. Bayona, R. Compan�o� , M. Granados, C. Leal and statistical method36 for the sum of each variability. If the M. Prat, J. Chromatogr. A, 1997, 788, 1. sampling strategy was not optimized against these ecological 18 M. Yoo and T. Kajiwara, Marine Fouling, 1983, 4, 11 (in parameters of mussel samples, the weight of variability Japanese). 19 T. Kajiwara, Y. Ura and N. Ito, Fisheries Sci., 1978, 44, 949 included in one analysed value was estimated to be (in Japanese). ±11.9–50.1% for TBT and 10.5–133.8% for TPT. The weight 20 N. P. Wilkins, K. Fujino and E. M. Gosling, Biol. J. Linn. Soc., of variability included in one analysed value using our optim- 1983, 20, 365.ized sampling strategy was estimated to be 16.2% for TBT 21 K. D. Daskalakis, Mar. Pollut. Bull., 1996, 32, 794. and 9.1% for TPT of the analysed value against the variability 22 R. B. Laughlin and W. French, Environ. Toxicol. Chem., 1988, due to analytical error only. We will be able to detect a 7, 1021. 23 R. F. Lee, Mar. Environ. Res., 1991, 32, 29. statistically significant diVerence of more than 20.6% for TBT 24 R.F. Lee, Mar. Ecol.-Prog. Ser., 1988, 46, 33. and 14.1% for TPT among the values of tissue concentration. 25 J. Capuzzo, J. Farrington, P. Rantamaki, C. CliVord, B. Judging from our study, the advantages of the use of blue Lancaster, D. Leavitt and X. Jia, Mar. Environ. Res., 1989, 28, mussels as exposure indicators may be summarized as follows: 489. (i) they permit the detection of contaminants that are present 26 H. Hummel, J. UniOudeGroeneveld, J. Nieuwenhuize, J. Liere, in ultra-trace levels or have no detectable concentration in R. Bogaards and L. Wolf, Sci. Total Environ., 1990, 92, 155. 27 P. B. Lobel and D. A. Wright, Mar. Pollut. Bull., 1982, 13, 320. seawater; (ii) they allow biological or ecological identification 28 A. J. S. Hawkins, E. Navarro and J. Iglesias, Mar. Biol., 1990, of potentially harmful chemicals in the environment; (iii) they 105, 197. can provide a rough approximation of chemical contamination 29 N. S. Fisher, J. Teyssie�, S. Fowler and W. Wang, Environ. Sci. and temporal and spatial comparisons of such contamination; Technol., 1996, 30, 3232. and (iv) they are stable environmental samples for storage and 30 K. Iida and M. Ogura, Annual Report of Kanagawa Institute of retrospective analysis.11 The limitations are summarized as Environmental Science, Kanagawa Institute of Environmental Science, Yokosuka, 1993, vol. 16, p. 45 (in Japanese). follows: (i) the dependence on the natural distribution of J. Environ. Monit., 1999, 1, 243–250 24931 A. L. Mallet and C. E. Carver, J. Exp. Mar. Biol. Ecol., 1993, ORCA 71, ed. G. Lauenstein and A. Cantillo, National Oceanic 170, 89. and Atmospheric Administration, Maryland, 1993. 32 D. M. Lowe, P. N. Salkeld and M. R. Carr, J. Mar. Biol. Assoc. 35 P. Lobel, S. Belkhode, S. Jackson and H. Longerich, Mar. UK, 1994, 74, 225. Environ. Res., 1991, 31, 163. 33 Y. Koshikawa, Y. Serizawa and A. Otsuki, IEEE Oceans 36 G. Christian, Analytical Chemistry, Wiley, New York, 4th edn., Proceedings, 1991, 2 (October 1–3), 1049. 1986, Section 3.8. 34 NOAA Technical Memorandum: Sampling and Analytical Methods of the National Status and Trends Program Benthic Surveillance and Mussel Watch Projects Analytical Protocols 1984–1992, NOS Paper 8/09669D 250 J. Environ. Monit., 1999, 1, 243&n

ISSN:0960-7919    

DOI:10.1039/a809669d

出版商:RSC    

年代:1999

数据来源: RSC