EN ISO 11269-1:2012

Soil quality - Determination of the effects of pollutants on soil flora - Part 1: Method for the measurement of inhibition of root growth (ISO 11269-1:2012)

ISO 11269-1:2012 describes a method for the determination of the effects of contaminated soils or contaminated samples on the root elongation of terrestrial plants.
This method is applicable to soils, soil materials, compost, sludge, waste or chemical testing. It is applicable to the comparison of soils of known and unknown quality and to the measurement of effects of materials (compost, sludge, waste) or chemicals deliberately added to the soil.
The method is not intended to be used as a measure of the ability of the soil to support sustained plant growth.

Bodenbeschaffenheit - Bestimmung der Wirkungen von Schadstoffen auf die Bodenflora - Teil 1: Verfahren zur Messung der Wurzelwachstumshemmmung (ISO 11269-1:2012)

Dieser Teil von ISO 11269 beschreibt ein Verfahren zur Bestimmung der Wirkungen von verunreinigten Böden oder verunreinigten Proben auf das Wurzellängenwachstum von Landpflanzen.
Dieses Verfahren ist anwendbar auf Böden, Bodenmaterialien, Kompost, Klärschlamm, Abfälle oder die Prüfung von Chemikalien. Es ist anwendbar für den Vergleich von Böden bekannter und unbekannter Beschaffenheit und für die Messung der Wirkungen von Materialien (Kompost, Schlamm, Abfälle) oder Chemikalien, die dem Boden kontrolliert zugegeben werden.
Das Verfahren ist nicht dafür vorgesehen, als Maß für die Fähigkeit eines Bodens, Pflanzenwachstum nachhaltig zu ermöglichen, verwendet zu werden.

Qualité du sol - Détermination des effets des polluants sur la flore du sol - Partie 1: Méthode de mesurage de l'inhibition de la croissance des racines (ISO 11269-1:2012)

L'ISO 11269-1:2012 décrit une méthode de détermination des effets de sols contaminés ou d'échantillons contaminés sur l'allongement des racines de végétaux terrestres.
Cette méthode est applicable aux sols et matériaux assimilés ainsi qu'aux composts, boues, déchets ou substances chimiques. Elle est applicable à la comparaison de sols de qualité connue et inconnue et au mesurage des effets de matériaux (composts, boues, déchets) ou de substances chimiques délibérément ajoutés au sol.
Cette méthode n'est pas destinée à être utilisée pour mesurer la capacité du sol à permettre la croissance assistée de végétaux.

Kakovost tal - Vpliv onesnaževal na talno floro - 1. del: Metoda za merjenje zaviranja rasti korenin (ISO 11269-1:2012)

Ta del standarda ISO 11269 opisuje metodo za ugotavljanje vpliva onesnaženih tal ali onesnaženih vzorcev na rast korenin kopenskih rastlin. Ta metoda se uporablja za tla, materiale v tleh, kompost, blato, odpadke ali kemijska preskušanja. Uporablja se za primerjavo tal znane in neznane kakovosti ter za merjenje vpliva materialov (kompost, blato, odpadki) ali kemikalij, ki se namenoma dodajo tlom. Ta metoda ni namenjena za merjenje, koliko so tla primerna za trajno rast rastlin.

General Information

• Status: Published
• Publication Date: 11-Dec-2012
• Withdrawal Date: 29-Jun-2013
• ICS: 13.080.30 - Biological properties of soils
• Technical Committee: CEN/TC 444 - Environmental characterization
• Drafting Committee: CEN/TC 444/WG 4 - Biological characterization
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 12-Dec-2012
• Due Date: 16-Jun-2013
• Completion Date: 12-Dec-2012
• Ref Project: SIST EN ISO 11269-1:2013 - Soil quality - Determination of the effects of pollutants on soil flora - Part 1: Method for the measurement of inhibition of root growth (ISO 11269-1:2012)

Overview - EN ISO 11269-1:2012 (Soil quality - Inhibition of root growth)

EN ISO 11269-1:2012 specifies a standardized phytotoxicity test for measuring the effect of contaminated soils, soil materials or added substances on the root elongation of terrestrial plants. The method compares root growth in a test soil or test mixtures (soil + compost, sludge, waste or chemicals) against a control (reference or standard soil) under controlled laboratory conditions. It is intended for ecotoxicological assessment and comparison of soil quality, not as a measure of long‑term crop productivity.

Keywords: EN ISO 11269-1:2012, soil quality, root growth inhibition, phytotoxicity test, ecotoxicological testing.

Key topics and technical requirements

• Test principle: Pregerminated seeds are grown in test and control substrates; root lengths are measured and statistically compared. Shoot height can be recorded as corroborative data.
• Applicable materials: Soils, soil materials, compost, sewage sludge, wastes, and chemicals added to soil.
• Test design elements:
  • Selection of test plants and pregermination procedures.
  • Use of reference soil, standard soil or site-specific control soil.
  • Preparation of pots/vessels and controlled growing conditions (environmental control to ensure reproducibility).
  • Methods for incorporating chemicals or materials into soil (annex guidance included).
• Data expression: Results can be reported as ECx (effective concentration reducing root elongation by x%) and NOEC (no-observed-effect concentration); mixtures often expressed as % dry mass.
• Validity and reporting: The standard defines validity criteria, measurement procedures, and required test report information.
• Supporting guidance: Annexes cover water‑holding capacity measurement, reference substance results, incorporation methods and example data.

Practical applications and target users

• Environmental testing laboratories performing soil quality and ecotoxicological assessments.
• Regulators and compliance bodies evaluating contaminated land, waste disposal on agricultural land, or sludge application.
• Remediation consultants and soil scientists assessing phytotoxic risks during brownfield reclamation, landfill capping or waste reuse.
• Research institutions studying pollutant impacts on terrestrial plant early growth.

Benefits: provides a reproducible, year‑round laboratory method for rapid screening of phytotoxic effects and for comparing soils or amendments.

Related standards

• ISO 11269-2 (emergence and early growth of higher plants)
• ISO 10381-6 (soil sampling for microbiological assessment)
• ISO 10390, ISO 10694, ISO 11260, ISO 11268-1/2, ISO 11277, ISO 11465, ISO/TS 20281 (referenced normative methods used alongside EN ISO 11269-1).

Use EN ISO 11269-1:2012 when you need a standardized, reproducible test for root growth inhibition to inform risk assessment, regulatory decisions or remediation planning for contaminated soils and soil amendments.

Standards Content (Sample)

SLOVENSKI STANDARD
01-maj-2013
1DGRPHãþD
SIST ISO 11269-1:2001
Kakovost tal - Vpliv onesnaževal na talno floro - 1. del: Metoda za merjenje
zaviranja rasti korenin (ISO 11269-1:2012)
Soil quality - Determination of the effects of pollutants on soil flora - Part 1: Method for
the measurement of inhibition of root growth (ISO 11269-1:2012)
Bodenbeschaffenheit - Bestimmung der Wirkungen von Schadstoffen auf die Bodenflora
- Teil 1: Verfahren zur Messung der Wurzelwachstumshemmmung (ISO 11269-1:2012)
Qualité du sol - Détermination des effets des polluants sur la flore du sol - Partie 1:
Méthode de mesurage de l'inhibition de la croissance des racines (ISO 11269-1:2012)
Ta slovenski standard je istoveten z: EN ISO 11269-1:2012
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN ISO 11269-1
NORME EUROPÉENNE
EUROPÄISCHE NORM
December 2012
ICS 13.080.30
English Version
Soil quality - Determination of the effects of pollutants on soil
flora - Part 1: Method for the measurement of inhibition of root
growth (ISO 11269-1:2012)
Qualité du sol - Détermination des effets des polluants sur Bodenbeschaffenheit - Bestimmung der Wirkungen von
la flore du sol - Partie 1: Méthode de mesurage de Schadstoffen auf die Bodenflora - Teil 1: Verfahren zur
l'inhibition de la croissance des racines (ISO 11269-1:2012) Messung der Wurzelwachstumshemmmung (ISO 11269-
1:2012)
This European Standard was approved by CEN on 11 November 2012.

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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2012 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 11269-1:2012: E
worldwide for CEN national Members.

Contents Page
Foreword .3

Foreword
The text of ISO 11269-1:2012 has been prepared by Technical Committee ISO/TC 190 “Soil quality” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 11269-1:2012 by
Technical Committee CEN/TC 345 “Characterization of soils” the secretariat of which is held by NEN.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by June 2013, and conflicting national standards shall be withdrawn at
the latest by June 2013.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN/CENELEC Internal Regulations, the national standards organisations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 11269-1:2012 has been approved by CEN as a EN ISO 11269-1:2012 without any
modification.
INTERNATIONAL ISO
STANDARD 11269-1
Second edition
2012-03-01
Soil quality — Determination of the
effects of pollutants on soil flora —
Part 1:
Method for the measurement of inhibition
of root growth
Qualité du sol — Détermination des effets des polluants sur la flore du
sol —
Partie 1: Méthode de mesurage de l’inhibition de la croissance des racines
Reference number
ISO 11269-1:2012(E)
©
ISO 2012
ISO 11269-1:2012(E)
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISO’s
member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail [e-mail]
Web www.iso.org
Published in Switzerland
ii © ISO 2012 – All rights reserved

ISO 11269-1:2012(E)
Contents Page
Foreword .iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Test plants . 3
6 Materials . 3
6.1 Test vessels . 3
6.2 Soil . 3
7 Equipment . 5
8 Reference substance . 5
9 Procedure . 5
9.1 Experimental design . 5
9.2 Preparation of pots . 6
9.3 Pregermination of the seeds . 6
9.4 Growing conditions . 6
9.5 Test duration . 6
9.6 Measurements . 7
10 Expression of results and data . 7
11 Validity criteria . 7
12 Test report . 7
Annex A (informative) Recommended method for measuring the water-holding capacity of the soil. 9
Annex B (informative) Results of tests performed on reference substances .10
Annex C (informative) Example of results obtained with boric acid using sand as the substrate . 11
Annex D (informative) Recommended methods for the incorporation of chemicals into soils .12
Annex E (informative) Recommended methods for the incorporation of compost, sludge or waste
into soils .13
Annex F (informative) Example of seedlings of winter barley collected at the end of the test after
removal from artificial soil .15
Bibliography .16
ISO 11269-1:2012(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International
Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 11269-1 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4,
Biological methods.
This second edition cancels and replaces the first edition (ISO 11269-1:1993), which has been technically revised.
ISO 11269 consists of the following parts, under the general title Soil quality — Determination of the effects of
pollutants on soil flora:
— Part 1: Method for the measurement of inhibition of root growth
— Part 2: Effects of contaminated soil on the emergence and early growth of higher plants
iv © ISO 2012 – All rights reserved

ISO 11269-1:2012(E)
Introduction
Chemical analysis of soil samples or waste materials to be disposed on soil, together with ecotoxicological
testing, provides substantial evidence of the suitability of the soil for arable production, or gives information
on the potential environmental risk resulting from the disposal of wastes such as sewage sludge on farmland.
There is also a need to assess the quality of the soil after reclamation of industrial sites and colliery tips or
when capping landfill sites. As the ability of the soil to grow crops is the main criterion, a rapid-growth test has
been developed, based on seedling growth in controlled environmental conditions.
Two major prerequisites of a phytotoxicity test are that it provides consistently reliable results and that it can be
used at any time of the year. It is therefore essential that seeds be grown in a controlled environment to ensure
optimal growing conditions which can be maintained for any number of tests, producing reproducible results
over a long period of time.
The test method described in this part of ISO 11269 can be used to compare soils, to monitor changes in their
activity or to determine the effect of added chemicals or materials (compost, sludge, waste).
INTERNATIONAL STANDARD ISO 11269-1:2012(E)
Soil quality — Determination of the effects of pollutants on soil
flora —
Part 1:
Method for the measurement of inhibition of root growth
1 Scope
This part of ISO 11269 describes a method for the determination of the effects of contaminated soils or
contaminated samples on the root elongation of terrestrial plants.
This method is applicable to soils, soil materials, compost, sludge, waste or chemical testing. It is applicable
to the comparison of soils of known and unknown quality and to the measurement of effects of materials
(compost, sludge, waste) or chemicals deliberately added to the soil.
The method is not intended to be used as a measure of the ability of the soil to support sustained plant growth.
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 10381-6, Soil quality — Sampling — Part 6: Guidance on the collection, handling and storage of soil under
aerobic conditions for the assessment of microbiological processes, biomass and diversity in the laboratory
ISO 10390, Soil quality — Determination of pH
ISO 10694, Soil quality — Determination of organic and total carbon after dry combustion (elementary analysis)
ISO 10930, Soil quality — Measurement of the stability of soil aggregates subjected to the action of water
ISO 11260, Soil Quality — Determination of effective cation exchange capacity and base saturation level using
barium chloride solution
ISO 11268-1, Soil quality — Effects of pollutants on earthworms — Part 1: Determination of acute toxicity to
Eisenia fetida/Eisenia andrei
ISO 11268-2, Soil quality — Effects of pollutants on earthworms — Part 2: Determination of effects on
reproduction to Eisenia fetida/Eisenia andrei
ISO 11277, Soil quality — Determination of particle size distribution in mineral soil material — Method by
sieving and sedimentation
ISO 11465, Soil quality — Determination of dry matter and water content on a mass basis — Gravimetric method
ISO/TS 20281, Water quality — Guidance on statistical interpretation of ecotoxicity data
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
contaminant
substance or agent present in the soil as a result of human activity
ISO 11269-1:2012(E)
3.2
test mixture
mixture of test soil or test material (compost, sludge, waste or chemical) with control soil
3.3
radicle
portion of the plant embryo which develops into the primary root
3.4
hypocotyl
portion of the axis of an embryo or seedling situated between the cotyledons (seed leaves) and the radicle
3.5
reference soil
uncontaminated site-specific soil (e.g. collected in the vicinity of a contaminated site) with similar properties
(nutrient concentrations, pH, organic carbon content and texture) as the test soil
3.6
standard soil
field-collected soil or artificial soil whose main properties (e.g. pH, texture, organic matter content) are within
a known range
[1] [2] 1)
EXAMPLE Euro-soils , artificial soil , LUFA soil.
NOTE The properties of standard soils can differ from the test soil.
3.7
control soil
reference or standard soil used as a control and as a medium for preparing dilution series with test soil or test
material (e.g. compost, sludge, waste, chemical)
NOTE Both the effective concentration (EC ) and the no-observed-effect concentration (NOEC) are expressed in
x
milligrams of test substance per kilogram (dry mass) of the test substrate. Soil mixtures are given in percent based on
soil dry mass.
3.8
effective concentration
ECx
effective concentration (dilution) of the test soil or test material (e.g. compost, sludge, waste, chemical) at which
root elongation is reduced by x % compared to the control
4 Principle
This method compares the root elongation of terrestrial plants in a test soil and/or a series of dilutions with a
control soil. This method may also be used for the testing of compost, sludge, waste or chemicals by applying
various concentrations of the material under investigation to a control soil.
Pregerminated seeds are exposed to the test material under controlled conditions. After the growth period,
the lengths of the roots of the test plants are compared with those of the control plants. Statistically significant
differences in the root lengths of seedlings grown in any test medium compared to the controls are indicative
of an effect.
NOTE Shoot height is also a useful parameter, and this can be measured in conjunction with root length to provide
additional or corroborative data.
1) Euro-soils, artificial soil and LUFA soil are examples of suitable products available commercially. This information is
given for the convenience of users of this document and does not constitute an endorsement by ISO of these products.
2 © ISO 2012 – All rights reserved

ISO 11269-1:2012(E)
5 Test plants
Winter barley (Hordeum vulgare L.), oat (Avena sativa L.) and wheat (Triticum aestivum L.) are the recommended
species. Other monocotyledonous plant species might be selected, e.g. plants with ecological or economic
significance in certain regions of the world, provided that the roots of these plants grow unhindered in sand and
in control soil under the conditions specified. Only plants that tolerate the properties of the test soils and test
conditions (besides their chemical contamination) should be selected. For example, a species sensitive to low
pH values should not be used for testing forest soils with low pH-values.
Seeds coated with insecticides and/or fungicides should not be used.
NOTE The methodology of this test can also be adapted for use with dicotyledonous species with straight roots,
which are easily measurable.
6 Materials
6.1 Test vessels
The test vessels shall be cylindrical, at least 8 cm in diameter and 11 cm in height, and shall have parallel sides
to ensure that the roots of seedlings are not restricted and do not encounter tapering side walls. The base of
the pots shall be perforated and covered with filter paper.
NOTE When filled to a height of 10 cm, the pots contain approximately 500 g of sand, 400 g of air-dried soil and 250
g of artificial soil.
6.2 Soil
6.2.1 Test soil
Some physical characteristics of the test soil can induce disturbances in root elongation such as heterogeneous
soil with big particles or clayey soil with a high water content. Therefore, the soil to be tested shall be passed
through a sieve with a 2 mm square mesh to remove coarse fragments. Furthermore, fine particles (<20 µm
according to ISO 11277) should not exceed 20 % of the dry mass.
Before the test, the soil is
...