CEN/TC 444 - Environmental characterization

Ecotoxicity tests can be applied to wastes to identify their potential hazardous properties with respect to the environment or to assess the risk related to a site-specific exposure scenario. This document provides guidance for the selection and use of ecotoxicity tests for both applications.
This document focuses on the following selected field of applications:
a)   Basic ecotoxicological characterization;
b)   Site-specific exposure scenario;
c)   Landfill management:
1)   monitoring of leachates;
2)   mineral waste going to non-controlled landfill sites.
d)   Re-use of waste:
1)   use of sludge in agriculture;
2)   use of mineral waste in road construction.
The user should be aware that other fields of application can also be covered by ecotoxicological testing not being in the scope of the document. The ecotoxicological assessment of waste within other scenarios might need the development of other test strategies.
Depending on the waste type and the assessment goal, relevant criteria are described for the selection of a test strategy and the suitable ecotoxicity test(s).
This document also provides guidance for individual ecotoxicity test protocols to meet the specific demands of waste testing (e.g. limitations, test design, confounding factors).The tests recommended represent a minimum test battery that may be accomplished by additional tests or even be replaced by others according to the waste, the intended use or protection goal envisaged.

This document specifies a method for the determination of the following elements in aqua regia, nitric acid or mixture of hydrochloric (HCl), nitric (HNO3) and tetrafluoroboric (HBF4)/hydrofluoric (HF) acid digests of soil, treated biowaste, waste, sludge and sediment:
Aluminium (Al), antimony (Sb), arsenic (As), barium (Ba), beryllium (Be), bismuth (Bi), boron (B), cadmium (Cd), calcium (Ca), cerium (Ce), chromium (Cr), cobalt (Co), copper (Cu), dysprosium (Dy), erbium (Er), europium (Eu), gallium (Ga), gadolinium (Gd), germanium (Ge), gold (Au), hafnium (Hf), holmium (Ho), indium (In), iridium (Ir), iron (Fe), lanthanum (La), lead (Pb), lithium (Li), lutetium (Lu) magnesium (Mg), manganese (Mn), mercury (Hg), molybdenum (Mo), neodymium (Nd), nickel (Ni), palladium (Pd), phosphorus (P), platinum (Pt), potassium (K), praseodymium (Pr), rhodium (Rh), ruthenium (Ru), samarium (Sm), scandium (Sc), selenium (Se), silicon (Si), silver (Ag), sodium (Na), strontium (Sr), sulfur (S), tantalum (Ta), tellurium (Te), terbium (Tb), thallium (Tl), thulium (Tm), thorium (Th), tin (Sn), titanium (Ti), tungsten (W), vanadium (V), yttrium (Y), ytterbium (Yb), zinc (Zn) and zirconium (Zr).
The method is also applicable to other extracts or digests originating from, for example, DTPA extraction, fusion methods or total digestion methods, provided the user has verified the applicability.
The method has been validated for the elements given in Table A.1 (sludge), Table A.2 (compost) and Table A.3 (soil). The method is applicable for other solid matrices and other elements as listed above, provided the user has verified the applicability.
This method is also applicable for the determination of major, minor and trace elements in aqua regia and nitric acid digests and in eluates of construction products (EN 17200[22]).
NOTE            Construction products include e.g. mineral-based products; bituminous products; metals; wood-based products; plastics and rubbers; sealants and adhesives; paints and coatings.

This document specifies an empirical method for the simultaneous direct determination of the fluorine, chlorine, bromine, and sulfur content in environmental solid matrices by oxidative pyrohydrolytic combustion at (1 050 ± 50) °C, followed by ion chromatography. The method is applicable for the determination of concentrations ≥ 10 mg/kg of each element based on dry matter. The upper limit and exact concentration range covered depends on the blank levels of the instrumentation and the capacity of the chromatographic separation column used for determination.
NOTE 1   Simultaneous determination of iodine content is possible but currently not validated.
NOTE 2   Other detection methods can be applied if validated.

This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Folsomia candida Willem by dermal and alimentary uptake. This document also provides information on how to use this method for testing substances under temperate conditions.
The chronic test described is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, industrial, agricultural or other sites of concern and waste materials.
The method is not applicable to volatile substances, i.e. substances for which H (Henry's constant) or the air/water partition coefficient is greater than 1, or for which the vapour pressure exceeds 300 Pa at 25 °C.

This document provides guidance to the characterization of waste. It applies to all types of waste, with unknown or partially known composition, by giving examples of EN standards dedicated to waste characterization and analytical methods for parameters not covered by standards. Some requirements concerning the determination of inorganic elements and organic substances content in waste are given to achieve approximately 90 % or the highest possible mass.
In case information on the origin or on the composition of the waste is given by the owner of the waste, it might be sufficient to follow only part of this document to complete missing knowledge about the waste.

This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Enchytraeus sp. by dermal and alimentary uptake in a chronic test. It is applicable to soils and soil materials of unknown quality, for example, from contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern and waste materials.
This document provides information on how to use this method for testing substances under temperate conditions.
The method is not applicable to substances, for which the air/soil partition coefficient is greater than 1, or to substances for which the vapour pressure exceeds 300 Pa at 25 °C.
NOTE      No provision is made in the test method for monitoring the persistence of the substance under test.

This document specifies one of the methods for evaluating the habitat function of soils and determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei by dermal and alimentary uptake. This chronic test is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, agricultural or other sites concerned, and waste materials.
This method is designed mainly for determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei. Technical information is also provided on how to use Eisenia fetida/andrei for testing chemicals under tropical conditions (see Annex A). Finally, this method also includes technical information on how to use it with other environmentally relevant earthworm species: e.g. Dendrodrilus rubidus and Aporrectodea caliginosa (see Annexes B and C).
This method does not apply to substances for which the air/soil partition coefficient is greater than one, or to substances with vapour pressure exceeding 300 Pa, at 25 °C. This method does not take into account the persistence of the substance during the test.

This European standard specifies a method for the differentiated determination of the organic carbon content (TOC400) which is released at temperatures up to 400 °C, the residual oxidizable carbon (ROC) (including e.g. lignite (brown coal), hard coal, charcoal, black carbon, soot) and the inorganic carbon (TIC900) which is released at temperatures up to 900 °C.
The basis is the dry combustion to CO2 in a in the presence of oxygen using using temperatures ranging from 150°C to 900 °C in dry solid samples of soil, soil with anthropogenic admixtures and solid waste (see Table 1) with carbon contents of more than 1 g per kg (0,1 % C) (per carbon type in the test portion).

This document specifies the crucial steps of a quantitative real-time polymerase chain reaction (qPCR) method to quantify the abundance of specific mRNA molecules extracted from Daphnia magna.
The method allows the identification of molecular responses to exposures for potentially toxic substances through the analysis of the abundance of specific mRNA molecules. In this document, the central genes involved in reproductive and toxic responses are included.
NOTE   The selection of genes can be adapted to specific exposure conditions, for example, exposure to known toxic substances, by adding genes known to respond to a specific insult.
The present method allows for rapid, robust and sensitive detection of molecular responses and can be used to analyse the toxic effects of water leachates from soil and waste. The method gives information of the concentration of a substance or test-liquid at which toxic effects begin to occur prior to observations of reproductive or toxic effects at higher levels of organization, which reduces the need for the use of safety factors in toxicity assessment.
The method is useful in several types of risk assessment. In this document, the genes studied are appropriate for the assessment of the risks when recycling materials and for the classification of waste, but the method can be adapted to other types of risk assessment by including other genes.

ISO 19204:2017 describes in a general way the application of the soil quality TRIAD approach for the site-specific ecological risk assessment of contaminated soils. In detail, it presents in a transparent way three lines of evidence (chemistry, ecotoxicology and ecology) which together allow an efficient, ecologically robust but also practical risk assessment of contaminated soils. This procedure can also be applicable to other stress factors, such as acidification, soil compaction, salinization, loss of soil organic substance, and erosion. However, so far, no experience has been gained with these other applications. Therefore, this document focuses on soils contaminated by chemicals.
NOTE 1       This document focuses on ecological risk assessment. Thus, it does not cover human health end points.
In view of the nature of this document, the investigation procedure is described on a general level. It does not contain details of technical procedures for the actual assessment. However, this document includes references relating to technical standards (e.g. ISO 15799, ISO 17616) which are useful for the actual performance of the three lines of evidence.
In ecological risk assessment, the effects of soil contamination on the ecosystem are related to the intended land use and the requirements that this use sets for properly functioning soil. This document describes the basic steps relating to a coherent tool for a site-specific risk assessment with opportunities to work out site-specific details.
ISO 19204:2017 can also be used for the evaluation of clean-up operations, remediation processes or management measures (i.e. for the evaluation of the environmental quality after having performed such actions).
NOTE 2       This document starts when it has already been decided that an ecological risk assessment at a given site needs to be performed. In other words, the practical performance of the soil quality TRIAD and the evaluation of the individual test results will be described. Thus, nothing will be said about decisions whether (and if yes, how) the results of the assessment are included in soil management measures or not.
NOTE 3       The TRIAD approach can be used for different parts of the environment, but this document focuses mostly on the soil compartment. Comparable documents for other environmental compartments are intended to be prepared in addition (e.g. the terrestrial aboveground compartment) in order to perform a complete site assessment, based on the same principles and processes.

This document is one of the family of standards (ISO 15799, ISO 19204) providing guidance on the characterization of soils and soil materials in relation to their retention and habitat functions and uses. It is appropriate to use it in conjunction with the two other standards in this family. It provides guidance on the choice and evaluation of tests applied for ecotoxicological characterization of soils and soil materials. Recommendations for test strategies with respect to the protection of ground and surface waters and the maintenance of the habitat function of soil are included. The tests recommended represent a minimum test battery that can be complemented by additional tests, or even be replaced by others, according to the intended uses or protection goals envisaged. The effect values indicated in this document do not refer to regulation but represent the lowest level at which an adverse effect is considered likely to occur.

This document is one of a family of International Standards providing guidance on soils and soil materials in relation to certain functions and uses including conservation of biodiversity. It applies in conjunction with these other standards. It provides guidance on the selection of experimental methods for the assessment of the ecotoxic potential of soils and soil materials (e.g. excavated and remediated soils, refills, embankments) with respect to their intended use and possible adverse effects on aquatic and soil dwelling organisms.
NOTE       This is a reflection of the maintenance of the habitat and retention function of the soil. In fact, the methods listed in this document are suitable for usage in a TRIAD approach, i.e. for an ecological assessment of potentially contaminated soils (see ISO 19204).
This document does not cover tests for bioaccumulation.
The ecological assessment of uncontaminated soils with a view to natural, agricultural or horticultural use is not within the scope of this document. Such soils can be of interest if they can serve as a reference for the assessment of soils from contaminated sites.
The interpretation of results gained by applying the proposed methods is not in the scope of this document.

This document specifies an instrumental method for the routine determination of pH within the range pH 2 to pH 12 using a glass electrode in a 1:5 (volume fraction) suspension of soil, sludge and treated biowaste in either water (pH in H2O), in 1 mol/l potassium chloride solution (pH in KCl) or in 0,01 mol/l calcium chloride solution (pH in CaCl2).
This document is applicable to all types of air-dried soil and treated biowaste samples.
NOTE       For example, pretreated in accordance with ISO 11464 or EN 16179 or EN 15002.

This document specifies a method for quantitative determination of the concentration of selected alcohols with low boiling point in liquid waste and pasty waste by gas chromatography with flame ionization detection after static headspace extraction.
Under the conditions specified in this document, a limit of application of 20 mg/kg, expressed on dry matter for pasty waste and expressed on raw waste for liquid waste, can be achieved.

This document specifies a method for sampling and handling free-living nematodes from terrestrial field soils as a prerequisite for using them as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms).
This document applies to all terrestrial biotopes in which nematodes occur. The sampling design of field studies in general is specified in ISO 18400-101.
This document is not applicable to aquatic nematodes because of differences in the sample matrix (e.g. water column). Methods for some other soil organism groups such as earthworms, collembolans enchytraeids or macro-invertebrates are covered in ISO 23611-1, ISO 23611-2, ISO 23611-3 and ISO 23611-5.
This document does not cover the pedological characterization of the site which is highly recommendable when sampling soil invertebrates. ISO 10390, ISO 10694, ISO 11272, ISO 11274, ISO 11277, ISO 11461 and ISO 11465 include suitable procedures for measuring pH, particle size distribution, C/N ratio, organic carbon content and water-holding capacity.

This document specifies two methods for the determination of total organic carbon (TOC) in sludge, treated biowaste, soil and waste samples containing more than 0,1 % carbon in relation to the dry mass (dm).
NOTE   This method can also be applied to other environmental solid matrices, provided the user has verified the applicability.

This document specifies different methods for quantitative determination of 16 polycyclic aromatic hydrocarbons (PAH) (see Table 2) in soil, sludge, treated biowaste, and waste, using GC-MS or HPLC-UV-DAD/FLD covering a wide range of PAH contamination levels (see Table 2).
NOTE   The method can be applied to sediments provided that validity is demonstrated by the user.
When using fluorescence detection, acenaphthylene cannot be measured.
[Table 2 -Target analytes of this document]
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used for the extraction of the sample and the clean-up of the extract.
Under the conditions specified in this document, the lower limit of application from 10 μg/kg (expressed as dry matter) for soils, sludge and biowaste to 100 μg/kg (expressed as dry matter) for solid waste can be achieved. For some specific samples (e.g. bitumen) the limit of 100 μg/kg cannot be reached.
Sludge, waste and treated biowaste can differ in properties as well as in the expected contamination levels of PAH and presence of interfering substances. These differences make it impossible to describe one general procedure. This document contains decision tables based on the properties of the sample and the extraction and clean-up procedure to be used.
The method can be applied to the analysis of other PAH not specified in the scope, provided suitability is proven by proper in-house validation experiments.
Sampling is not part of this standard. In dependence of the materials, the following standards need to be considered, e.g. EN 14899, ISO 5667-12 and EN ISO 5667-13.

This document specifies an operationally defined method for the direct determination of organically bound halogens (chlorine, bromine and iodine) adsorbed and occluded to the sample matrix. AOX being a methodologically defined parameter, it is essential that the procedure is applied without any modification.
This document is intended for analysis of sludge, treated biowaste or soil in concentrations ranging from 5 mg/kg dry matter. The upper limit and exact concentration range covered depend on the instrumentation used for determination.
NOTE This method can also be applied to other environmental solid matrices, provided the user has verified the applicability.

This document describes a method to assess the bioaccumulation of chemicals in snails, i.e. concentrations of metal(loid)s (ME) or organic compounds [e.g. polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs)] accumulated in their tissues.
This document presents how to prepare snails for caging in situ for 28 days, the in situ test design and then how to collect and prepare the snails until conservation and further analysis. If a kinetic study of accumulation is necessary, sampling of snails at different time-points during exposure is possible as well [13],[19],[22].
This document excludes analytical methods. Preparation (extraction and mineralization) of the samples and quantification of chemicals are not in the scope of the present document.
The method is applicable for soils under different uses (agricultural, industrial, residential, forests, before and after remediation, on potentially contaminated sites, etc.) and waste materials [8],[10], preferably with vegetation and/or humus cover.
The method is applicable subject to certain limits of temperature (frost-free period, i.e. mainly from April to October in temperate region).
Optionally (see Annex I), the method can be used in the laboratory to evaluate the accumulation of contaminants [and optionally, the sum of excess of transfer (SET) index for ME, PAH, PCB] of snails exposed only to soil.

This document specifies an up-flow percolation test (PT) which is applicable in compliance testing to determine the leaching behaviour of inorganic and non-volatile organic substances from granular solids with potential for use as construction material. The test is not suitable for substances that are volatile under ambient conditions. The granular solids are subjected to percolation with water as a function of liquid to solid ratio under specified percolation conditions. The method is a once-through column leaching test.
NOTE 1   Volatile organic substances include the low molecular weight substances in mixtures such as mineral oil.
This up-flow percolation test is performed under specified test conditions for granular solids with potential for use as construction material and does not necessarily produce results that mimic specific intended use conditions. This test method produces eluates, which can subsequently be characterized by physical, chemical and ecotoxicological methods according to existing standard methods. The results of eluate analysis are presented as a function of the liquid/solid ratio. The test results enable the distinction between different leaching behaviour.
NOTE 2   It is not always possible to adjust test conditions simultaneously for inorganic and organic substances. Test conditions can also vary between different groups of organic substances. Test conditions for organic substances are generally more stringent than those for inorganic substances. The test conditions are generally described in a way that they fit testing organic substances and are also applicable to inorganic substances depending on the set-up.
NOTE 3   For ecotoxicity testing, eluates representing the release of both inorganic and organic substances are needed. In this document, ecotoxicological testing is meant to include also genotoxicological testing.
NOTE 4   Granular solid waste materials with a low hydraulic conductivity that can cause detrimental pressure build-up are not supposed to be subjected to this test.
NOTE 5   This procedure is generally not applicable to solids that are easily biologically degradable and solids reacting with the leachant, leading to, for example, excessive gas emission or excessive heat release, impermeable hydraulically bound solids or solids that swell in contact with water.
Granular solid waste materials without potential for beneficial use are excluded from the scope.
NOTE 6   Granular solid waste materials without potential for beneficial use can be materials with gas generation or biodegradation during a potential reuse scenario.
This test is applicable to types of granular solid waste of which the general long-term leaching behaviour is known based on previous investigations.
In this document the same test conditions as for EN 16637 3 (CEN/TC 351/WG 1) are applied in order to allow full comparability of testing construction products and waste derived construction products to avoid double testing. The EN 16637 3 test results are eligible in the context of testing granular solids with potential for use as construction material as well.
NOTE 7   If a leaching test according to EN 16637 3 has been performed, additional EN 17516 testing does not need to be carried out.
NOTE 8   The relative standard deviations for inorganic and organic substances are relatively high which is due to low concentration levels in the eluates (see Annex F).

This document describes the necessary steps to be performed before carrying out ecotoxicity tests on wastes. The purpose of this document is to provide guidance on the taking of the sample, transport, storage of wastes and to define preparation, for the determination of ecotoxicological properties of wastes under the conditions specified in this document by biological testing either as raw wastes or water extracts from wastes. Sample preparation for other applications (e.g. assessment of waste effects on aquatic and terrestrial organisms in a disposal scenario) is not considered.
Specifying a test battery to characterize ecotoxicological properties of wastes is not in the scope of this document.
This document is applicable to solid and liquid wastes.

This document specifies a method for the determination of total dissolved solids (TDS) in water and eluates (see Annex A), provided they are not volatile under the conditions specified or that they do not release water molecules from hydration. It applies to water and eluates containing more than 100 mg/l of total dissolved solids. Samples with lower amounts of dissolved solids can be analysed by repetition of the drying step.

This document specifies the measurement of explosives and related nitrocompounds (as given in Table 1) in soil and soil materials. This document is intended for the trace analysis of explosives and related compounds by liquid chromatography–tandem mass spectrometry (LC-MS/MS). Generally, LC-MS/MS measurement shows the lower LOQ (limit of quantification) for each compound in Table 1 than using high-performance liquid chromatography (HPLC) with UV-detection (see Annex B and Annex C).
Under the conditions specified in this document, concentrations as low as 0,005 mg/kg to 0,014 mg/kg-dry matter can be determined, depending on the substance. Similar compounds, in particular various nitroaromatics, by-products and degradation products of explosive compounds can be analysed using this method provided that the applicability is checked on a case-by-case basis.

This document specifies a method for the determination of the loss on ignition (LOI) at 550 °C. The dry matter is determined according to EN 15934.
This method applies to the determination of loss on ignition of sediment, sludge, treated biowaste, soil and waste.
NOTE   The loss on ignition is often used as an estimate for the content of organic matter in the sample. Inorganic substances or decomposition products (e.g. H2O, CO2, SO2, O2) are released or absorbed and some inorganic substances are volatile under the reaction conditions.

This document specifies the determination of Cr(VI) in solid waste material and soil by alkaline digestion and ion chromatography with spectrophotometric detection. This method can be used to determine Cr(VI)-mass fractions in solids higher than 0,1 mg/kg.
NOTE       In case of reducing or oxidising waste matrix no valid Cr(VI) content can be reported.

This document provides guidance on the selection and application of screening methods for assessing soil quality and waste characterization, including distribution of target parameters in soil and soil‑like material. The aim of this document is to set up criteria as to when the different kind of screening methods can be applied for the analysis of a certain parameter in soil, including soil‑like material, and waste, and which steps are required to prove their suitability.
This document does not recommend any particular screening method but confirms the principles of their selection and application.

This document specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and individual chemical substances on the reproduction of the oribatid mite Oppia nitens by dermal and alimentary uptake. This chronic (28-day) test is applicable to soils and soil materials of unknown quality (e.g., contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern and waste materials). This method is not intended to replace the earthworm or Collembola tests since it represents another taxonomic group (= mites; i.e., arachnids), nor the predatory mite test since this species represents a different trophic level and ecological niche.
Effects of substances are assessed using standard soil, preferably a defined artificial soil substrate. For contaminated soils, the effects are determined in the test soil and in a control soil. According to the objective of the study, the control and dilution substrate (dilution series of contaminated soil) should be either an uncontaminated soil with similar properties to the soil sample to be tested (reference soil) or a standard soil (e.g., artificial soil).
Information is provided on how to use this method for testing substances under temperate conditions.
This document is not applicable to substances for which the air/soil partition coefficient is greater than 1, or to substances with vapour pressure exceeding 300 Pa at 25 °C.
NOTE    The stability of the test substance cannot be assured over the test period. No provision is made in the test method for monitoring the persistence of the substance under test.

This document specifies two methods for digestion of soil, treated biowaste, sludge and waste by the use of an aqua regia digestion.
Digestion with aqua regia will not necessarily accomplish total decomposition of the sample. The extracted analyte concentrations may not necessarily reflect the total content in the sample but represent the aqua regia soluble metals under the condition of this test procedure. It is generally agreed that for environmental analysis purposes, the results are fit for the intended purpose to protect the environment.
This document is applicable for the following elements:
Aluminium (Al), antimony (Sb), arsenic (As), barium (Ba), beryllium (Be), boron (B), cadmium (Cd), calcium (Ca), chromium (Cr), cobalt (Co), copper (Cu), iron (Fe), lead (Pb), magnesium (Mg), manganese (Mn), mercury (Hg), molybdenum (Mo), nickel (Ni), phosphorus (P), potassium (K), selenium (Se), silver (Ag), sodium (Na), strontium (Sr), sulfur (S), tellurium (Te), thallium (Tl), tin (Sn), titanium (Ti), vanadium (V), and zinc (Zn).
This document can also be applied for the digestion of other elements, provided the user has verified the applicability.

This document specifies a simple method for the extraction of only phospholipid fatty acids (PLFA) from soils.

This document specifies three methods for the digestion of soil, treated biowaste, sludge and waste by the use of an acid mixture composed of hydrochloric (HCl), nitric (HNO3) and tetrafluoroboric (HBF4) or hydrochloric (HCl), nitric (HNO3) and hydrofluoric (HF) acid as the digestion solution.
Digestion with these acids is effectively considered as a total decomposition of the sample. For a broad range of samples the extracted analyte concentrations will reflect the total content in the sample.
This document is applicable for the following elements:
Aluminium (Al), antimony (Sb), arsenic (As), barium (Ba), beryllium (Be), cadmium (Cd), calcium (Ca), chromium (Cr), cobalt (Co), copper (Cu), iron (Fe), lead (Pb), magnesium (Mg), manganese (Mn), mercury (Hg), molybdenum (Mo), nickel (Ni), phosphorus (P), potassium (K), selenium (Se), silver (Ag), sodium (Na), strontium (Sr), sulfur (S), tellurium (Te), thallium (Tl), tin (Sn), titanium (Ti), vanadium (V), and zinc (Zn).
This document can also be applied for the digestion of other elements, provided the user has verified the applicability.

The present document specifies a method for direct extraction of DNA from soil samples to analyse the abundance and composition of microbial communities by various techniques of molecular biology including real-time quantitative PCR (qPCR). This method is mainly dedicated to agricultural and forest soils. This method can possibly not be suitable for soils rich in organic matter (e.g. peat soils) or soils heavily polluted with organic pollutants or heavy metals.
The direct extraction of DNA from soil samples provides unique insight into the α- and β-diversity of microbial communities. Next-generation sequencing of amplicons obtained by PCR (polymerase chain reaction) amplification of soil DNA constitutes a promising domain which will in the near future contribute to the development of routine tools to monitor microbial communities in soil environments.

This document specifies methods for quantitative determination of seven selected polychlorinated biphenyls (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 and PCB180) in soil, sludge, sediment, treated biowaste, and waste using GC-MS and GC-ECD (see Table 2).
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used for the extraction of the sample and the clean-up of the extract.
Under the conditions specified in this document, lower limit of application from 1 μg/kg (expressed as dry matter) for soils, sludge and biowaste to 10 μg/kg (expressed as dry matter) for solid waste can be achieved. For some specific samples the limit of 10 μg/kg cannot be reached.
Sludge, waste and treated biowaste may differ in properties, as well as in the expected contamination levels of PCB and presence of interfering substances. These differences make it impossible to describe one general procedure. This document contains decision tables based on the properties of the sample and the extraction and clean-up procedure to be used.
NOTE   The analysis of PCB in insulating liquids, petroleum products, used oils and aqueous samples is referred to in EN 61619, EN 12766-1 and EN ISO 6468 respectively.
The method can be applied to the analysis of other PCB congeners not specified in the scope, provided suitability is proven by proper in-house validation experiments.

2020-02-13 - waiting TC decision to skip FV.

This document provides guidance on developing and using conceptual site models (CSMs) through the various phases of investigation, remediation (if required), and any subsequent construction or engineering works.
It describes what CSMs are, what they are used for and what their constituents are. It stresses the need for an iterative and dynamic approach to CSM development.
This document is intended to be used by all those involved in developing CSMs and by those who rely on using them such as regulators, landowners, developers, and the public (and other relevant parties). Ideally, this includes representatives from all phases of the investigative and remedial processes, for example, preliminary assessment, detailed investigation, baseline human health and environmental risk assessments, and feasibility study, and, any subsequent construction or engineering work.
NOTE 1    This document is applicable whenever the presence of "potentially harmful" or "hazardous" substances are present irrespective of whether they are naturally occurring or present due to human activity (i.e. are "contaminants").
NOTE 2    Although most of the principles described for developing CSMs in this document can apply to other domains, such as groundwater resources management, the present document is specifically written for the management of potentially contaminated sites or known contaminated sites.

This document describes an approach for the validation of physico-chemical analytical methods for environmental solid matrices and water.
The guidance in this document addresses the initial description of the method and two different validation approaches, in increasing order of complexity. These are:
a)   method development, if the method is developed by the laboratory, or conditions of adoption, if the method is a standardized protocol adopted by the laboratory;
b)   validation at the level of single laboratories (within-laboratory validation);
c)   method validation at the level of several laboratories (between-laboratory or inter-laboratory validation), with a focus on methods that are sufficiently mature and robust to be applied not only by a few expert laboratories but by laboratories operating at the routine level.
The concept is strictly hierarchical, i.e. a method shall fulfil all criteria of within-laboratory validation before it can enter the validation protocol of the between-laboratory.
This document is applicable to the validation of a broad range of quantitative physico-chemical test methods for the analysis of water (including drinking water, surface water, groundwater, waste water, marine water), and of solid environmental matrices, such as soil, sludge, liquid and solid waste, sediment and biota. It is intended for standardized protocols adopted by a laboratory, and either for test methods aiming at substances that have recently become of interest or for test methods applying recently developed technologies.
The minimal requirements that are indispensable for the characterization of the fitness for the intended purpose of an analytical method are: selectivity, precision, trueness, performances characteristics and measurement uncertainty. The aim of validation is to prove that these requirements are met.
In this document after the definitions (Clause 3) and description of the principles (Clause 4) a toolbox is given describing the relevant performance characteristics in the validation process.
Clause 7 and 8 focus on the within laboratory validation process (V1) and Clause 9 on the interlaboratory validation process (V2). Clause 7 and 8 describe largely the same processes, but differ in approach for establishing the LOQ.
Reporting of the results of the validation studies is addressed in Clause 10.

ISO 17512-2:2011 specifies a rapid screening method for evaluating the habitat function of soils based on the avoidance behaviour of springtails.
The test is a rapid method that reflects the bioavailability of contaminants in natural soils and substances spiked into soils to Folsomia candida. In both cases, it is possible to establish a dose-response-relationship. The avoidance behaviour of the springtails is the measurement endpoint of the test. This test is not intended to replace the Collembola reproduction test.

This document specifies a method for the measurement of several hydrolase activities (arylamidase, arylsulfatase, β-galactosidase, α-glucosidase, β-glucosidase, N-acetyl-glucosaminidase, acid, alkaline and global phosphatases, urease) simultaneously (or not) in soil samples, using colorimetric substrates. Enzyme activities of soil vary seasonally and depend on soil chemical, physical and biological characteristics. This method can be applied either to detect harmful effects on soil enzyme activities derived from toxic substances or other anthropogenic agents in contaminated soils against a control soil, or to test chemicals.

ISO 15685:2012 specifies a rapid method for the determination of the potential rate of ammonium oxidation and inhibition of nitrification in soils. This method is suitable for all soils containing a population of nitrifying microorganisms. It can be used as a rapid screening test for monitoring soil quality and quality of wastes, and is suitable for testing the effects of cultivation methods, chemical substances [except volatiles i.e. H > 1 (Henry's constant)], extracts of biosolids and pollution in soils.

1 Scope
This International Standard specifies a test method for determining the activity of active aerobic, heterotrophic microbial biomass in soils. This method is applicable to the monitoring of soil quality and to the evaluation of the ecotoxic potential of soils and soil materials. It is also applicable for soils sampled along contamination gradients in the field and to soils that are contaminated experimentally in the field or in the laboratory.

This document describes a method to compare the quality of soils by determining the fatty acid composition of the leaves of plant species grown in these soils.
This method does not make it possible to determine an optimal value of the Omega-3 index and, therefore, cannot be used to determine the intrinsic quality of a soil from a specific area (regarded as homogeneous). The method can only be used to compare the quality of soils between various areas.
This method is applicable to:
—          soils from contaminated sites;
—          amended soils;
—          soils after remediation;
?      soil with waste products (e.g. slurry, manure, sludge or composts).
Alternatively, the quality of soils can be assessed by determining the Omega-3 index of Lactuca sativa seedlings grown in these soils under controlled conditions (i.e. phytotronic chamber) and by comparing these values to those obtained from control soils (see Annex B).

ISO 17512-1:2008 specifies a rapid screening method for evaluating the habitat function of soils and the influence of contaminants and chemicals on earthworm behaviour.
The sublethal test is a rapid method that reflects the bioavailability of contaminant mixtures in natural soils and substances spiked into soils to Eisenia fetida and Eisenia andrei. The avoidance behaviour of the worms is the measurement endpoint of the test. This test is not intended to replace the earthworm reproduction test.
Two different designs (a two section unit and a six section unit) have been developed and successfully applied. Both designs are applicable to either single-concentration (e. g. for assessing the quality of a field soil) or multi-concentration (e. g. for assessing the toxicity of a spiked chemical) tests. In both cases, the earthworms are allowed to make the initial choice on which compartment, control and a treatment [in the two section test vessel between right and left side; in the six section test vessel between the (3 + 3) alternating compartments], to enter.

The purpose of ISO 29200:2013 is to describe a method for assessing genotoxic effects (chromosome breakage or dysfunction of the mitotic spindle) of soils or soil materials on the secondary roots of a higher plant: Vicia faba (broad bean). This method allows the assessment of genotoxicity (toxicity for genetic material) of soils and soil materials like compost, sludge, waste, fertilizing matters, etc. Two ways of exposure can be considered: a direct exposure of plants to the soil (or soil material) which is relevant for the real genotoxic potential and an exposure of plants to the water extract of the soil (or soil material). This last way of exposure to a leachate or an eluate allows the detection of the mutagens which are not adsorbed to soils and which may be transferred to aquatic compartments. Moreover, this test may be used to evaluate genotoxic effects of chemical substances and to waters, effluents, etc.

Provides guidance on the selection and conduct of appropriate test methods for the determination of biodegradation of organic chemicals in aerobic soils. Does not describe any specific test method.

ISO 18763:2016 describes a technique for determining the effects of soil and soil-related materials on the seed germination and early growth of higher plants. These endpoints are useful indicators for the assessment of the quality of a soil as a habitat for organisms. It is applicable to all soils in which soil organisms are active and may be used to evaluate:
-      the effects on plants due to toxicity of solid or liquid chemicals contaminating soil or materials (compost, sludge, waste) and chemicals added to soil;
-      the changes in the soil effect on plants after restoration measures.

ISO 14239:2017 specifies six suitable incubation systems for measuring the rates and extent of mineralization of organic compounds in soil by measurement of carbon dioxide (CO2) evolution. All incubation systems are applicable to soluble or insoluble compounds but choice of system depends on the overall purposes of the study.
ISO 14239:2017 does not apply to the use of such systems for material balance studies, which are often test-substance specific.

This document specifies a protocol to identify ecotoxicological test specimens (mainly invertebrates and plants) to the species level, based on the DNA barcoding technique. This protocol can be used by laboratories performing DNA barcoding in order to standardize both the wet-lab and data analysis workflows as much as possible, and make them compliant with community standards and guidelines.
This document does not intend to specify one particular strain for each test method, but to accurately document the species/strain which was used.
NOTE 1    This does not imply that DNA barcoding is performed in parallel to each test run, but rather regularly (e.g. once a year, such as reference substance testing) and each time a new culture is started or new individuals are added to an ongoing culture.
This document does not aim at duplicating or replacing morphological-based species identifications. On the contrary, DNA barcoding is proposed as a complementary identification tool where morphology is inconclusive, or to diagnose cryptic species, in order to ensure that the results obtained from different ecotoxicological laboratories are referring to the same species or strain.
This document is applicable to identifications of immature forms which lack morphological diagnostic characters (eggs, larvae, juveniles), as well as the streamline identification of specimens collected in field monitoring studies, where large numbers of organisms from diverse taxa are classified.
NOTE 2    In principle, all species regularly used in ecotoxicological testing can be analysed by DNA barcoding. Besides the earthwoms Eisenia fetida and E. andrei, further examples for terrestrial species are Lumbricus terrestris, L. rubellus, Allolobophora chlorotica, Aporrectodea rosea, and A. caliginosa, Dendrodrilus rubidus, Enchytraeus albidus, and E. crypticus (Haplotaxida); Folsomia candida, F. fimetaria, Proisotoma minuta, and Sinella curviseta (Collembola); Hypoaspis aculeifer and Oppia nitens (Acari); Aleochara bilineata and Poecilus cupreus (Coleoptera); Scathophaga stercoraria, Musca autumnalis (Diptera) or Pardosa sp. (Arachnida). Nematodes or snails and even plants can also be added to this list.

This document specifies a chronic test method for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Hypoaspis aculeifer by ? mainly ? alimentary uptake. This method is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, industrial, agricultural or other sites under concern and waste materials (e.g. dredged material, municipal sludge from a wastewater treatment plant, composed material, or manure, especially those for possible land disposal). The reproduction (= number of juveniles) is the measured parameter of the test. The test reflects the bioavailability of a mixture of contaminants in natural soils (contaminated site soils) to a species which represents a trophic level which is not covered by other ISO standards. This test is not intended to replace the earthworm (see ISO 11268-2) or Collembola (see ISO 11267) reproduction tests since this species belongs not only to a different trophic group but also a different taxonomic group (= mites; i.e. arachnids) than those used usually.
Effects of substances are assessed using a standard soil, preferably a defined artificial soil substrate. For contaminated soils, the effects are determined in the soil to be tested and in a control soil. Depending on the objective of the study, the control and dilution substrate (dilution series of contaminated soil) are either an uncontaminated soil comparable to the soil to be tested (reference soil) or a standard soil (e.g. artificial soil).
This document provides information on how to use this method for testing samples (soils or substances) under temperate conditions.
This document is not applicable to substances for which the air/soil partition coefficient is greater than one, or to substances with vapour pressure exceeding 300 Pa at 25 °C.
NOTE       The stability of the test substance cannot be ensured over the test period. No provision is made in the test method for monitoring the persistence of the substance under test.

This International Standard gives guidance on the selection and method of appropriate tests for the determination of biodegradation of organic chemicals in soil samples under anaerobic conditions.
NOTE For methods intended for tests in the framework of the registration of chemicals, an OECD Guideline on soil degradation gives useful guidance on additional test requirements.