SIST EN ISO 22030:2011

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Bodenbeschaffenheit - Biologische Verfahren - Chronische Toxizität in höheren Pflanzen (ISO 22030:2005)Qualité du sol - Méthodes biologiques - Toxicité chronique sur les plantes supérieures (ISO 22030:2005)Soil quality - Biological methods - Chronic toxicity in higher plants (ISO 22030:2005)13.080.30Biološke lastnosti talBiological properties of soilsICS:Ta slovenski standard je istoveten z:EN ISO 22030:2011SIST EN ISO 22030:2011en,fr,de01-november-2011SIST EN ISO 22030:2011SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN ISO 22030
June 2011 ICS 13.080.30 English Version
Soil quality - Biological methods - Chronic toxicity in higher plants (ISO 22030:2005)
Qualité du sol - Méthodes biologiques - Toxicité chronique sur les plantes supérieures (ISO 22030:2005)
Bodenbeschaffenheit - Biologische Verfahren -Chronische Toxizität in höheren Pflanzen (ISO 22030:2005) This European Standard was approved by CEN on 10 June 2011.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
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Reference numberISO 22030:2005(E)© ISO 2005
INTERNATIONAL STANDARD ISO22030First edition2005-02-01Soil quality — Biological methods — Chronic toxicity in higher plants Qualité du sol — Méthodes biologiques — Toxicité chronique sur les plantes supérieures
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ISO 22030:2005(E) © ISO 2005 – All rights reserved
iii Contents Page Foreword.iv Introduction.v 1 Scope.1 2 Normative references.1 3 Terms and definitions.2 4 Principle.4 5 Materials.4 5.1 Test plants.4 5.2 Soil and soil storage.4 5.2.1 General.4 5.2.2 Test soil.5 5.2.3 Control soil.5 5.3 Reference substance.6 6 Apparatus.6 7 Methods.8 7.1 Experimental design.8 7.2 Pre-test.8 7.3 Preliminary test (range-finding).8 7.4 Final test.9 7.5 Preparation of the pots.9 7.6 Preparation of the seeds.9 7.7 Growth conditions.9 7.8 Start of the test.10 7.9 Handling during the test.10 7.9.1 Number of plants and thinning out.10 7.9.2 Watering.10 7.9.3 Rearrangement of test vessels.10 7.9.4 Pollination.10 7.9.5 Reporting.11 7.10 Endpoint measurements.11 7.10.1 Seedling emergence.11 7.10.2 Harvest at day 14.11 7.10.3 Final harvest.11 7.11 Summary and timetable of the test.12 8 Validity criteria.13 9 Assessment of the results.13 9.1 Presentation of measured data.13 9.2 Statistical analysis.14 9.2.1 Preliminary test.14 9.2.2 Final test.14 9.2.3 NOEC (no observed effect concentration) approach.14 9.2.4 ECx (effect concentration) approach.14 10 Test report.15 Annex A (informative)
Testing of chemicals in soil.16 Bibliography.17
ISO 22030:2005(E) iv
ISO 22030:2005(E) © ISO 2005 – All rights reserved
v Introduction This International Standard describes a procedure for evaluating the quality of soils of different origin carrying unknown contaminations. The method, slightly modified, can also be used to measure the toxicity of known chemicals incorporated into soil.
The evaluation of the inhibition and chronic toxicity is based on emergence, vegetative growth and reproductive capacity of at least two species of higher plants.
This International Standard is based on: a) results of the research project “Development of a chronic bioassay using higher plants”, sponsored by the German Ministry for Education and Research (BMBF), Bonn [3], and b) discussions within the joint project “Ecotoxicological Test Batteries” forming part of the BMBF Joint Research Group “Processes for the Bioremediation of Soil” [10].
INTERNATIONAL STANDARD ISO 22030:2005(E) © ISO 2005 – All rights reserved
1 Soil quality — Biological methods — Chronic toxicity in higher plants WARNING — Contaminated soils can contain unknown mixtures of toxic, mutagenic or otherwise harmful chemicals or infectious microorganisms. Occupational health risks can arise from dust or evaporated chemicals during handling and incubation. Furthermore, test plants can absorb chemicals from the soil and safety measures should also be considered when handling these test plants. 1 Scope This International Standard describes a method for determining the inhibition of the growth and reproductive capability of higher plants by soils under controlled conditions. Two species are recommended: a rapid-cycling variant of turnip rape (Brassica rapa CrGC syn. Rbr) and oat (Avena sativa). The duration of test should be sufficient to include chronic endpoints that demonstrate the reproductive capability of the test plants. By using natural test soils, e.g. from contaminated sites or remediated soils, and by comparing the development of the test plants in these soils with reference or standard control soils, the test can be used to assess soil quality, especially the function of the soil as a habitat for plants. Annex A describes modifications allowing use of the chronic plant assay for the testing of chemicals incorporated into soil. By preparing a dilution series of a test substance in standard control soils, the same endpoints can be measured to assess the chronic toxicity of chemicals. This 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 0,013 3 Pa at 25 °C. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 11268-1:1993, Soil quality — Effects of pollutants on earthworms (Eisenia fetida) — Part 1: Determination of acute toxicity using artificial soil substrate ISO 11268-2:1998, Soil quality — Effects of pollutants on earthworms (Eisenia fetida) — Part 2: Determination of effects on reproduction ISO 11269-2, Soil quality — Determination of the effects of pollutants on soil flora — Part 2: Effects of chemicals on the emergence and growth of higher plants ISO 15176:2002, Soil quality — Characterization of excavated soil and other soil materials intended for re-use ISO 15799, Soil quality — Guidance on the ecotoxicological characterization of soils and soil materials ASTM D1076:2002, Standard Specification for Rubber-Concentrated, Ammonia Preserved, Creamed, and Centrifuged Natural Latex SIST EN ISO 22030:2011

ISO 22030:2005(E) 2
NOTE 1 Biomass is expressed as dry mass per plant or, if needed, as dry mass per pot. NOTE 2 During the test period, some of the test plants can reach different growth stages and their water content can differ when the plants are harvested. Thus the dry mass better represents the biomass produced during the growth period. 3.3 concentration mass of test substance per amount of soil NOTE Concentration is expressed as a mass fraction, in milligrams per kilogram (mg/kg) of dry soil. 3.4 contaminant substance or agent present in the soil as a result of human activity [ISO 15176:2002] 3.5 control soil uncontaminated substrate, used as a control and as medium for preparing dilution series with test soils or chemicals, that allows the growth of healthy plants NOTE Either artificial or natural standard or reference soils can be used, if unhindered growth of the test plants in these soils can be expected. In any case, differences in nutrient levels between a test soil and a control soil can affect the dose-response pattern. For example, a control soil much richer in nutrients than a test soil can result in a false positive result (i.e. the test soil appears to have a “toxic” effect on the growth of the test plants). If a control soil is poorer in nutrients than a test soil, hormesis (see 3.9) can be expected at low soil-mixture ratios, or even an inverse dose response relationship, if nutrient supply becomes the main effect. This International Standard does not provide numerical values for the nutrients. 3.6 effect concentration
ECx concentration (mass fraction) of a test chemical or the percentage (mass fraction) of a test soil at which a given endpoint is inhibited by x % compared to the control NOTE The effect concentration is expressed in milligrams per kilogram. When chemicals are tested, the ECx is expressed as mass of the test substance per dry mass of soil; when soils are tested, the ECx is expressed as a percentage of test soil dry mass per soil mixture dry mass. 3.7 emergence development of a seedling contained within a seed, ending the latent period NOTE It is expressed as the percentage of seedlings which emerge from test pots as compared with the control pots. SIST EN ISO 22030:2011

ISO 22030:2005(E) © ISO 2005 – All rights reserved
3 3.8 habitat function ability of soils/soil materials to serve as a habitat for microorganisms, plants, soil-living animals and their interactions (biocenosis) [ISO 15799] 3.9 hormesis improvement of seedling emergence, growth or survival (or other response of the test plants) at low concentrations of chemicals or mixtures of soil that are toxic when applied at higher levels in comparison to the control [1] 3.10 lowest observed effect concentration LOEC lowest tested concentration (mass fraction) of a test substance in soil at which a statistically significant effect on a given endpoint (p < 0,05) compared with the control is observed cf. NOEC (3.11) NOTE Analogously, the term LOEC is used for the lowest tested mixture ratio of a test soil in a reference or a standard control soil at which a statistically significant effect is observed. The LOEC is expressed as mass of the test substance per mass of dry soil or, in the latter case, as percentage of test-soil dry mass per soil-mixture dry mass. All test concentrations above the LOEC have a harmful effect equal or greater than that observed at the LOEC. If this condition cannot be satisfied, an explanation should be given for how the LOEC and NOEC have been selected. 3.11 no observed effect concentration NOEC test substance concentration (mass fraction) or soil mixture ratio immediately below the LOEC, which when compared to the control has no statistically significant effect (p < 0,05)
cf. LOEC (3.10) 3.12 reference soil uncontaminated site-specific soil (e.g. collected in the vicinity of a contaminated site) with properties (nutrient concentrations, pH, organic carbon content and texture) similar to the test soil 3.13 soil mixture ratio ratio of the dry mass of test soil to the dry mass of reference/control soil NOTE It is expressed as a percentage. 3.14 standard soil field-collected soil or artificial soil whose main properties (e.g. pH, texture, organic matter content) are within a known range EXAMPLES Euro soils, artificial soil. NOTE The properties of standard soils may differ from those of the test soil. SIST EN ISO 22030:2011

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1) Seeds and Wisconsin Fast Plants kits are suitable products supplied by the Carolina Biological Supply Company, Burlington, NC, USA. This information is given for the convenience of users of this International Standard and does not constitute an endorsement by ISO of the product named. Equivalent products may be used if they can be shown to lead to the same results. SIST EN ISO 22030:2011

ISO 22030:2005(E) © ISO 2005 – All rights reserved
5 The chronic plant test can be used to assess the toxic potential of natural soils from a variety of contaminated sites. It can also be used to assess the quality of remediated soils. The selected soils should have pH values after sieving within a range that is not toxic to the test plants, e.g. between 5,0 and 7,5 for Brassica rapa and Avena sativa. Test soils shall be passed through a sieve of mesh 4 mm to 5 mm square to remove coarse fragments, and mixed thoroughly. If necessary, soil may be air-dried without heating before sieving. Storage of test soils should be as short as possible. Storage at approximately 4 °C using containers that minimize losses of soil contaminants by volatilization and sorption to the container walls is recommended. 5.2.2 Test soil The water-holding capacity shall be measured for all soil mixtures used in the test. Additionally, test soils should be characterized by:  texture (sand, silt, clay);  pH value;  salinity;  organic carbon;  total and water-soluble amounts of potassium, nitrogen and phosphorus. Soil pH should not be corrected. Measurements of soil contaminants (heavy metals, hydrocarbons, pesticides, explosives, PCBs and others) are not mandatory. It should be checked whether the test soil sucks water via wicks sufficiently (see 5.4). Water repellency or poor water transport can occur with very sandy soils, soils highly contaminated with hydrocarbons or even with soils of high clay content that tend to compact even when these soils have a high water-holding capacity (determined after initially submerging the soils). To assure functioning of the watering system, a pre-test including all soils selected for the test and replicated twice should be performed to decide whether wick watering is sufficient or manual watering is required. NOTE For the time being, pH limits for plant species other than turnip rape and oat cannot be given. It is matter of future research to systematically test more plants on a variety of soils. Furthermore, tolerance limits for texture, salinity or other soil properties cannot yet be given for different plant species. 5.2.3 Control soil Either artificial, reference or standard soils may be used as control substrate. If reference soils from uncontaminated areas near a contaminated site are available, they should be treated and characterized like the test soils. In addition, to verify that a reference soil does not carry toxic contaminants, chemical analysis of the expected contaminants shall be carried out. If toxic contamination or unusual soil properties cannot be ruled out, standard control soils should be given preference. Standard soils should be uncontaminated, nutrient-poor natural or artificial soils. If a natural soil is used, its organic matter content should not exceed 5 %. Fine particles (< 20 µm) should not exceed 20 %. Alternatively, artificial soil in accordance with reference [6] and ISO 11268-2 may be used, regardless of its higher organic matter content. However, the organic matter contents of the test and control soil should be as close to each other as possible. The artificial soil consists of the following components (percentage based on dry mass):  10 % sphagnum peat [air-dried and finely ground (2 mm ± 1 mm)];  20 % kaolin clay (kaolinite content preferably above 30 %);  approximately 69 % (depending on the amount of CaCO3 needed) air-dried industrial quartz sand (predominantly fine sand with more than 50 % mass fraction of particle size 0,05 mm to 0,2 mm). SIST EN ISO 22030:2011

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