kSIST-TS FprCEN/TS 17459:2020

Construction products - Assessment of release of dangerous substances - Determination of ecotoxicity of construction product eluates

This document specifies a test procedure that combines horizontal leaching tests with ecotoxicity tests for the assessment of eluates of construction products.
The test procedure in this document is established for construction products with an organic content of > 50 % by mass. Mineral-based construction products are covered in the following case: 1) the liquid or pasty product hardens in direct contact with soil or groundwater and 2) the used binder contains > 50 % organics by mass. For the covered products adequate validation data have been provided (see Annex A).
This document is not applicable for the assessment of terrestrial ecotoxicity of construction products.
NOTE   Terrestrial ecotoxicity tests for construction products are described in CEN/TR 17105.

Bauprodukte - Bewertung der Freisetzung von gefährlichen Stoffen - Bestimmung der Ökotoxizität von Eluaten aus Bauprodukten

Dieses Dokument legt ein Prüfverfahren fest, das horizontale Auslaugprüfungen mit Ökotoxizitätsprüfungen zur Bewertung der Eluate von Bauprodukten kombiniert.
Das Prüfverfahren in diesem Dokument wird für Bauprodukte mit einem organischen Massenanteil von > 50 % angegeben. Mineralische Bauprodukte sind im folgenden Fall abgedeckt: 1) Das flüssige oder pastöse Produkt erhärtet im direkten Kontakt mit dem Boden oder Grundwasser und 2) das verwendete Bindemittel enthält einen organischen Massenanteil von > 50 %. Für die abgedeckten Produkte wurden entsprechende Validierungsdaten bereitgestellt (siehe Anhang A).
Dieses Dokument ist nicht auf die Bewertung der terrestrischen Ökotoxizität von Bauprodukten anwendbar.
ANMERKUNG   Terrestrische Ökotoxizitätsprüfungen von Bauprodukten werden in CEN/TR 17105 beschrieben.

Gradbeni proizvodi - Ocenjevanje sproščanja nevarnih snovi - Določevanje ekotoksičnosti izlužkov gradbenih proizvodov

General Information

• Status: Not Published
• Public Enquiry End Date: 02-Feb-2020
• ICS: 13.020.99 - Other standards related to environmental protection
• ICS: 91.100.01 - Construction materials in general
• Technical Committee: NES - Dangerous substances
• Current Stage: 4020 - Public enquire (PE) (Adopted Project)
• Start Date: 04-Dec-2019
• Due Date: 22-Apr-2020
• Completion Date: 06-Feb-2020
• Directive: 305/2011 - Regulation (eu) No 305/2011 of the European Parliament and of the Council of 9 march 2011 laying down harmonised conditions for the marketing of construction products and repealing council directive 89/106/eec
• Mandate: M/366 - Concerning the execution of standardisation work for the development of horizontal standardised assessment methods for harmonized approcaches relating to dangerous substances under the construction products directive (CPD)
• Ref Project: FprCEN/TS 17459 - Construction products - Assessment of release of dangerous substances - Determination of ecotoxicity of construction product eluates

Standards Content (sample)

SLOVENSKI STANDARD
kSIST-TS FprCEN/TS 17459:2020
01-januar-2020
Gradbeni proizvodi - Ocenjevanje sproščanja nevarnih snovi - Določevanje
ekotoksičnosti izlužkov gradbenih proizvodov

Construction products - Assessment of release of dangerous substances - Determination

of ecotoxicity of construction product eluates

Bauprodukte - Bewertung der Freisetzung von gefährlichen Stoffen - Bestimmung der

Ökotoxizität von Eluaten aus Bauprodukten
Ta slovenski standard je istoveten z: FprCEN/TS 17459
ICS:
13.020.99 Drugi standardi v zvezi z Other standards related to
varstvom okolja environmental protection
91.100.01 Gradbeni materiali na Construction materials in
splošno general
kSIST-TS FprCEN/TS 17459:2020 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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kSIST-TS FprCEN/TS 17459:2020
FINAL DRAFT
TECHNICAL SPECIFICATION
prTS 17459
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
November 2019
ICS 91.100.01
English Version
Construction products - Assessment of release of
dangerous substances - Determination of ecotoxicity of
construction product eluates
Bauprodukte - Bewertung der Freisetzung von
gefährlichen Stoffen - Bestimmung der Ökotoxizität
von Eluaten aus Bauprodukten

This draft Technical Specification is submitted to CEN members for Vote. It has been drawn up by the Technical Committee

CEN/TC 351.

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, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and

United Kingdom.

Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are

aware and to provide supporting documentation.

Warning : This document is not a Technical Specification. It is distributed for review and comments. It is subject to change

without notice and shall not be referred to as a Technical Specification.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels

© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. prTS 17459:2019 E

worldwide for CEN national Members.
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Contents Page

European foreword ....................................................................................................................................................... 3

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms and definitions ................................................................................................................................... 6

4 Abbreviations ................................................................................................................................................... 8

5 Leaching procedures for ecotoxicological testing ............................................................................... 8

5.1 Suitable leaching tests and selection of fractions from leaching tests ........................................ 8

5.1.1 General ................................................................................................................................................................ 8

5.1.2 Dynamic Surface Leaching Test (DSLT) CEN/TS 16637-2 ................................................................ 9

5.1.3 Horizontal up-flow percolation test (CEN/TS 16637-3) ................................................................... 9

5.1.4 Alternative leaching tests............................................................................................................................. 9

5.2 Sampling, transport and storage of eluates ........................................................................................... 9

5.3 Pre-treatment of eluates............................................................................................................................ 10

6 Aquatic ecotoxicological testing ............................................................................................................. 11

6.1 Purpose of aquatic ecotoxicity testing .................................................................................................. 11

6.2 Selection of ecotoxicity tests and minimum test battery ............................................................... 11

6.3 Testing strategy and conditions ............................................................................................................. 12

6.4 Limitations of aquatic ecotoxicity tests ............................................................................................... 13

7 Assessment of biodegradability .............................................................................................................. 13

8 Test report and quality assurance procedure ................................................................................... 14

Annex A (informative) Results from a European round robin test “Ecotoxicological

characterisation of eluates from construction products” ............................................................. 15

Annex B (informative) Evaluation of test results............................................................................................. 19

Bibliography ................................................................................................................................................................. 21

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European foreword

This document (FprCEN/TS 17459:2019) has been prepared by Technical Committee CEN/TC 351

“Construction products: Assessment of release of dangerous substances”, the secretariat of which is held

by NEN.
This document is currently submitted to the Vote on TS.

This document has been prepared under a standardization request given to CEN by the European

Commission and the European Free Trade Association.
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Introduction

The regulatory and technical background for this FprCEN/TS 17459 is explained in CEN/TR 17105 [1].

General information on ecotoxicity assessment is also provided there. This Technical Specification

describes the assessment of ecotoxicological properties of eluates of construction products received from

horizontal leaching tests harmonized by CEN/TC 351.

Aquatic ecotoxicity tests can be applied to eluates of construction products which come into contact with

soil or water in their intended use as described in 3.2.2.2 Products relevant for soil, surface water or

groundwater of CEN/TR 16098:2010 [2] and in CEN/TS 16637-1. The test procedure specified in this

document covers construction products that fall under the scope of the leaching tests in the

CEN/TS 16637 series and have an organic content of > 50 % by mass. For the covered products adequate

validation data has been provided (see Annex A).

This document is recommended to Technical Committees for construction products (product TCs), if they

have been mandated to address ecotoxicity in their product standards or if they are interested to include

ecotoxicity in a dossier prepared in the context of qualifications for a “without testing” status. This

document is also recommended to EOTA in case there is a wish to include ecotoxicity testing in European

Assessment Documents.
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1 Scope

This document specifies a test procedure that combines horizontal leaching tests with ecotoxicity tests

for the assessment of eluates of construction products.

The test procedure in this document is established for construction products with an organic content

of > 50 % by mass. Mineral-based construction products are covered in the following case: 1) the liquid

or pasty product hardens in direct contact with soil or groundwater and 2) the used binder

contains > 50 % organics by mass. For the covered products adequate validation data have been provided

(see Annex A).

This document is not applicable for the assessment of terrestrial ecotoxicity of construction products.

NOTE Terrestrial ecotoxicity tests for construction products are described in CEN/TR 17105.

2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements 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.

EN 1484, Water analysis — Guidelines for the determination of total organic carbon (TOC) and dissolved

organic carbon (DOC)

EN 12457-1, Characterisation of waste — Leaching — Compliance test for leaching of granular waste

materials and sludges — Part 1: One stage batch test at a liquid to solid ratio of 2 l/kg for materials with

high solid content and with particle size below 4 mm (without or with size reduction)

EN ISO 5667-16:2017, Water quality — Sampling — Part 16: Guidance on biotesting of samples (ISO 5667-

16:2017)

EN ISO 6341, Water quality — Determination of the inhibition of the mobility of Daphnia magna Straus

(Cladocera, Crustacea) — Acute toxicity test (ISO 6341)

EN ISO 8692, Water quality — Fresh water algal growth inhibition test with unicellular green algae

(ISO 8692)

EN ISO 9408, Water quality — Evaluation of ultimate aerobic biodegradability of organic compounds in

aqueous medium by determination of oxygen demand in a closed respirometer (ISO 9408)

EN ISO 9439, Water quality — Evaluation of ultimate aerobic biodegradability of organic compounds in

aqueous medium — Carbon dioxide evolution test (ISO 9439)

EN ISO 11348-1, Water quality — Determination of the inhibitory effect of water samples on the light

emission of Vibrio fischeri (Luminescent bacteria test) — Part 1: Method using freshly prepared bacteria

(ISO 11348-1:2007)

EN ISO 15088:2008, Water quality — Determination of the acute toxicity of waste water to zebrafish eggs

(Danio rerio) (ISO 15088:2007)

EN ISO 20079:2006, Water quality — Determination of the toxic effect of water constituents and waste

water on duckweed (Lemna minor) — Duckweed growth inhibition test (ISO 20079:2005)

ISO 11350, Water quality — Determination of the genotoxicity of water and waste water —

Salmonella/microsome fluctuation test (Ames fluctuation test)
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ISO 13829, Water quality — Determination of the genotoxicity of water and waste water using the umu-

test

CEN/TS 16637-1:2018, Construction products: Assessment of release of dangerous substances — Part 1:

Guidance for the determination of leaching tests and additional testing steps

CEN/TS 16637-2, Construction products: Assessment of release of dangerous substances — Part 2:

Horizontal dynamic surface leaching test

CEN/TS 16637-3, Construction products: Assessment of release of dangerous substances — Part 3:

Horizontal up-flow percolation test
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at http://www.iso.org/obp
— IEC Electropedia: available at http://www.electropedia.org/
3.1
bioassay

technique for evaluating the biological effect, either qualitatively or quantitatively, of various substances

in water by means of changes in a specified biological activity

Note 1 to entry: Bioassay is a broader term than ecotoxicity test and covers also for instance biodegradability

tests.
[SOURCE: ISO 6107-8:1993/Amd 1:2001 [3], no number]
3.2
biodegradation

mineralization of organic compounds by bacteria and fungi to carbon dioxide, water and inorganic

compounds
3.3
control
mixture of control medium and organisms used in the test without test sample
3.4
control medium
combination of dilution water and/or nutrient medium used in the test
[SOURCE: EN ISO 20079:2005, 3.6]
3.5
dilution level

reciprocal value of the volume fraction of test sample in dilution water in which the test is conducted

EXAMPLE 250 ml of test sample in a total volume of 1 000 ml (volume fraction of 25 %) represents dilution

level D = 4.
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[SOURCE: EN ISO 15088:2008, 3.2, modified - “waste water” replaced by “test sample”]

3.6
dilution water
water added to the test sample to prepare a series of defined dilutions
Note 1 to entry: The composition of the water is specified in the specific test.
[SOURCE: EN ISO 20079:2006, 3.7]
3.7
ecotoxicity test

method for determining potential adverse effects to biological systems which a sample has an inherent

capacity to cause
3.8
effective concentration

concentration of the test sample at which an effect of x % is measured, if compared to the control

[SOURCE: ISO 20227:2017 [4], 3.1]
3.9
eluate
aqueous solution recovered from a leaching test
[SOURCE: CEN/TR 16110:2010 [5], 3.2]
3.10
leaching test

laboratory test during which a construction product is put into contact with a leachant under strictly

defined conditions for the determination of the release of substances into water
3.11
lowest ineffective dilution
dilution factor
LID

lowest ineffective dilution tested, expressed as dilution level D (3.5), at which no inhibition, or only effects

not exceeding the test-specific variability, are observed
[SOURCE: EN ISO 15088:2008, 3.5]
3.12
storage time

period of time between filling of the sample container and further treatment of the sample in the

laboratory, if stored under predefined conditions
[SOURCE: EN ISO 5667-3:2018 [6], 3.4]
3.13
test material
material to be tested
[SOURCE: ISO 17126:2005 [7], 3.3]
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4 Abbreviations
For the purposes of this document, the following abbreviations apply.
CEN/TC Technical Committee in CEN
CEN/TR Technical Report adopted by CEN
CEN/TS Technical Specification adopted by CEN
DOC dissolved organic carbon
DSLT dynamic surface leaching test
EC effective concentration
EOTA European Organization for Technical Assessment
LID lowest ineffective dilution
TOC total organic carbon
5 Leaching procedures for ecotoxicological testing
5.1 Suitable leaching tests and selection of fractions from leaching tests
5.1.1 General

This Technical Specification is part of a modular horizontal approach and belongs to the analytical step

(CEN/TS 16637-1:2018, Figure 2, number 6). This approach was adopted in CEN/TC 351. “Horizontal”

means that the methods can be used for products with certain properties as described in

CEN/TS 16637-1. “Modular” means that a test standard developed in this approach concerns a specific

step in assessing a property and not the whole “chain of measurement” (from sampling to analyses).

Differing from the modules for chemical analyses this Technical Specification also contains limited

modifications to the leaching step (CEN/TS 16637-1:2018, Figure 2, number 5) as detailed below.

A leaching method shall be selected using guidance given in CEN/TS 16637-1. Eluates from the DSLT

according to CEN/TS 16637-2 and column test according to CEN/TS 16637-3 are suitable for ecotoxicity

tests on the basis of current experience.

The selection of fractions from leaching tests for ecotoxicity tests depends on the purpose of the testing

approach. Generally, for many construction products the maximum effect is expected to occur with the

first elution steps. It is often also practicable to test the first two fractions, because organic substances

are not always stable during long immersion periods.

An alternative option for the selection of fractions is to define an indicative parameter (e.g. TOC or

substances of interest) which is analysed in each eluate fraction. The two fractions with the highest

amount of the indicative parameter are combined and tested.

For construction products with continuously releasing compounds, those fractions with the longest

contact time from an advanced stage of the leaching procedure can exhibit higher effects. By comparison

of effects from different fractions, the longer term behaviour of the construction products can be

assessed. The use of this option should be justified with an indicative parameter.

The results of ecotoxicity tests with the same product, but using a different leaching test are not

comparable due to the different test conditions.
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5.1.2 Dynamic Surface Leaching Test (DSLT) CEN/TS 16637-2

CEN/TS 16637-2 describes a tank test for monolithic construction products of > 40 mm edge length in all

directions and for plates or sheets with a surface area of > 100 cm exposed to the eluate. In the standard

procedure, this test is carried out for 64 days, while the eluate water is replaced at distinct time intervals

(after 6 h, 24 h, 2,25 d, 4 d, 9 d, 16 d, 36 d and 64 d).

In order to generate suitable eluates for ecotoxicity testing the DSLT shall be performed as follows:

— only the first two elution steps after 6 h and additional 18 h are carried out and both eluates are

unified for ecotoxicity testing;
2 2

— the lower limit of the liquid / surface area relation (L/A), corresponding to 20 l/m (2 ml/cm ), is

applied in order to maximize the concentration level in the eluates (smallest liquid to surface area).

NOTE These adjustments have been applied in the European round robin test for the ecotoxicological

characterization of construction products [8].

It is recommended to investigate eluates that represent other elution steps, if information on later stages

of leaching processes is desired.
5.1.3 Horizontal up-flow percolation test (CEN/TS 16637-3)

The horizontal up-flow percolation test is a method to determine the leaching behaviour of non-volatile

inorganic and organic substances from granular construction products (without or with size reduction).

The construction products 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.

Eluates from up-flow percolation tests according to CEN/TS 16637-3 shall be collected for ecotoxicity

testing until an L/S ratio of 2 l/kg is reached. Mass of test material and dimension of the applied columns

have to be adapted to obtain sufficient eluate volume for testing. Furthermore the relationship between

maximum grain size and minimal column dimension as specified in CEN/TS 16637-3 has to be fulfilled.

It is recommended to investigate eluates that represent higher L/S ratios, if information on later stages

of leaching processes is desired.
5.1.4 Alternative leaching tests

The conditions for using other leaching tests (indirect tests) are defined in CEN/TS 16637-1:2018,

Clause 7. From the alternative methods referred to in CEN/TS 16637-1 especially EN 12457-1 and

EN 16105 are known to generate suitable eluates for ecotoxicity tests on the basis of experience with

construction products.

EN 12457-1 is a one stage batch test with a liquid to solid ratio of 2 l/kg for materials with high solid

content and with particle size below 4 mm (without or with size reduction). Where CEN/TS 16637-3 is

used as the reference method, EN 12457-1 shall be used for quality assurance where applicable.

Products that come into contact with water only occasionally in their intended use may not be stable in a

test set up with long time immersion in water like foreseen in the reference method CEN/TS 16637-2.

For these products a leaching test with short immersion phases according to EN 16105 is recommended

instead. In this test the construction products are exposed to cycles of wetting and drying.

5.2 Sampling, transport and storage of eluates

The goal of sampling is to obtain a representative eluate and to supply it to the laboratory in the correct

manner. The eluates may be susceptible to changes as a result of physical, chemical or biological

reactions. The eluates should be preferably tested immediately after sampling to avoid changes in the

original composition as a result of physical and chemical reactions and/or biological processes. If this is

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not possible, e.g. because several bioassays are intended to be applied, samples shall be cooled down for

transport and storage in closed, dark glass bottles to temperatures between 2 °C and 8 °C and kept in the

dark or frozen down to ≤ −18 °C. The transport shall be realized in cool boxes with ice, frozen gel packs

or cooling elements. The time span between sampling and ecotoxicity tests (storage time) shall remain

within 48 h (cooled samples) or 2 months (frozen samples). For further information see EN ISO 5667-16

and related standards referred to there.

Eluates are not suitable for bioassays, if they have been treated to prevent biodegradation processes

during the leaching test. Preservatives such as biocides, acids or basic solutions that retard chemical and

biological activity shall not be added to the test samples. If samples are frozen, the DOC shall be

determined according to EN 1484 both before freezing and after thawing the sample.

The amount of eluates and number of replicate vessels depends on the test battery chosen and the test

design and should be decided beforehand. It is practicable to split the total volume of the eluate into

appropriate flasks. In this way, the different tests can be carried out independently from each other and

suitable test portions are available for potential repetitions. As a rough estimate about 100 ml to 200 ml

are needed for one test with one organism. For the algae test up to about 500 ml are required depending

on the size of the incubation vessels. Additional replicate samples for retesting the eluates in cases, where

the toxicity range has not been met, are required.

Frozen samples shall be thawed on the day of testing before use or overnight. When thawing on the day

of testing a warm water bath at a temperature not exceeding 25 °C, together with gentle shaking, shall be

used to avoid local overheating. When thawing overnight the sample shall be kept in the dark at a

temperature between 2 °C and 8 °C. A once thawed sample shall not be frozen again for subsequent

testing, but a replicate sample shall be used. An even distribution of all soluble and particulate

components shall be ensured (e.g. by gentle agitation or vigorous shaking).

Samples thawed with a microwave treatment are not suitable for biological tests as thawing in a

microwave oven modifies the test sample in an unacceptable way and is therefore forbidden.

NOTE It is possible to extend the duration of storage, if it has been proved that no modification of toxicity

occurred within the storage period (e.g. through comparison with the same ecotoxicity test immediately after

sampling and at the end of storage period).
5.3 Pre-treatment of eluates

In general, bioassays are carried out with the original sample. In accordance with EN ISO 5667-16 any

pre-treatment of the eluates shall be limited to such cases, where suspended solids resulting from

construction products disturb the performance of the bioassays.

When carrying out aquatic tests in the presence of particles causing severe problems, it is recommended

that the sample be allowed to settle for 30 min to 2 h. The required quantity of supernatant can be

sampled using a pipette. Only when large amounts of particulate matter interfere with the requirements

of test organisms, this shall be removed by centrifugation or filtration. Centrifugation (e.g. 10 min at

5 000 g) in general is preferred to filtration. Alternatively coarse particles may be separated by filtration

(>50 µm). The filter material shall be made of inert material (e.g. glass-fibre) and the filters shall be rinsed

with high-purity water prior to use to reduce the risk of contamination of the eluates. Filtration may be

carried out under pressure or with vacuum. Any method used for separation of particles shall be reported

in the test report.

When phase separation is restricted, e.g. due to the specific weight of the material, it may also be

supported by sieving the eluate through a (0,5 ± 0,2) mm stainless steel sieve and subsequent

centrifugation.

The eluate shall be allowed to settle for 30 min to 2 h after treatment. The required quantity of

supernatant can be sampled using a pipette.
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6 Aquatic ecotoxicological testing
6.1 Purpose of aquatic ecotoxicity testing

Aquatic ecotoxicity tests measure the water available fraction of the toxic components of the eluate.

When carrying out aquatic tests, attention should be focussed on possible confounding factors which

might interfere with the test results and make adaptations in the test schemes advisable. Differences in

the ionic composition might interfere with many of the test organisms; coloured or turbid samples might

interfere with algae as well as in the luminescent bacteria test and pH values might be outside the normal

range for many test organisms. In those cases tests should best be carried out twice: once according to

the standardized method to provide comparable results and once with an adaptation in the method to

provide more insight for the evaluation of results.

NOTE Guidance on suitable adaptations is given in EN ISO 5667-16:2017, Clause 9 “Impairment of test

performance”.
6.2 Selection of ecotoxicity tests and minimum test battery

It is generally accepted in environmental hazard assessment, that a test battery with organisms from

different trophic levels and combining different assessment end points, is required.

An appropriate test battery to be used in the aquatic ecotoxicity testing of construction product eluates

where applicable is detailed in the following tables (Table 1 and Table 2).

Table 1 — Minimum test battery for aquatic ecotoxicity tests on construction product eluates

Mandatory Expression of
Test species Standard End point
tests results
EC (30 min),
Luminescent bacteria (Vibrio
EN ISO 11348-1 Luminescence
fischeri)
LID
Algae
Minimum Pseudokirchneriella
EC (72 h), LID
EN ISO 8692 Growth
battery subcapitata or
Desmodesmus subspicatus
Crustacea
EC (48 h), LID
EN ISO 6341 Mobility
Daphnia magna
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Table 2 — Extended test battery in addition to the minimum test battery for aquatic ecotoxicity

tests on construction product eluates
Optional Expression of
Test species Standard End point
tests results
EC (48 h),
Danio rerio (zebrafish eggs) EN ISO 15088 Mortality
LID
umu-assay with Salmonella
typhimurium TA
Induction ratio
Extended
ISO 13829 Gene induction
1535/pSK1002
(24 h)
test battery
or,
Increase in the
increase of
Ames fluctuation test ISO 11350 number of
revertant wells
mutant colonies
6.3 Testing strategy and conditions
6.3.1 General

As a general guidance for testing strategy and conditions the guidance given in EN ISO 5667-16 should

be followed.
6.3.2 Additional blank samples

Additional blank vessels and eluates, which include all experimental steps (from dilution water over the

leaching vessels to the sampling vessels) shall be prepared in parallel and examined in the ecotoxicity

tests, in order to detect possible influence from the testing device such as the quality of theleachant,

contamination of the leaching vessels or the surrounding environment, the storage conditions etc.

Normally no ecotoxicity should be detectable in these additional blanks at a dilution level of 2 (50 % by

volume of the blank eluate and 50 % dilution water).
6.3.3 pH adjustment

pH of test mixtures (i.e. test sample diluted with control/dilution medium) may significantly differ from

pH of test sample according to the selected dilution range and according to buffer capacity of

control/dilution medium or test sample. A pH adjustment shall be performed according to the following

rules given in EN ISO 5667-16:
— samples with extreme pH values exceedi
...