Characterization of waste - Preparation of waste samples for ecotoxicity tests

This European Standard describes the necessary steps to be performed before carrying out ecotoxicity tests on wastes. The purpose of this European Standard 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 European Standard 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 European Standard.
This European Standard is applicable to solid and liquid wastes.

Charakterisierung von Abfällen - Herstellung von Abfallproben für ökotoxikologische Untersuchungen

Diese Europäischen Norm beschreibt die erforderlichen Arbeitsschritte, die durchzuführen sind, bevor
Ökotoxizitätsprüfungen an Abfällen durchgeführt werden. Der Zweck dieser Europäischen Norm ist, eine
Anleitung für Probenahme, Transport und Lagerung von Abfällen zu geben und die Vorbereitungen für die
Bestimmung ökotoxischer Eigenschaften von Abfällen festzulegen; diese Bestimmung erfolgt unter den in
dieser Europäischen Norm festgelegten Bedingungen durch biologische Prüfungen an Rohabfällen oder
wässrigen Extrakten aus Abfällen. Die Probenvorbereitung für andere Anwendungsweisen (z. B. Beurteilung
der Wirkungen von Abfällen auf Boden- und Wasserorganismen im Rahmen einer Entsorgung) wird nicht
berücksichtigt.
Die Festlegung einer Prüfbatterie zur Charakterisierung der ökotoxischen Eigenschaften von Abfällen liegt
nicht im Aufgabenbereich dieser Europäischen Norm.
Diese Europäische Norm ist auf feste und flüssige Abfälle anwendbar.

Caractérisation des déchets - Caractérisation des déchets - Préparation des échantillons de déchets en vue d'essais écotoxicologiques

Cette Norme européenne décrit toutes les étapes nécessaires a effectuer avant de réaliser des essais d’écotoxicité sur les déchets. Elle vise a fournir des directives pour l’échantillonnage, le transport et le stockage des déchets et a définir la préparation pour la détermination des propriétés écotoxicologiques des déchets, dans les conditions décrites dans cette Norme européenne, en utilisant des essais biologiques soit sur déchets bruts soit sur extraits aqueux de ces déchets. La préparation des échantillons pour d’autres applications (par exemple, l’évaluation des effets des déchets sur les organismes aquatiques ou terrestres dans un scénario d’élimination) n’est pas prise en considération.
La définition d’une batterie d’essais pour la caractérisation des propriétés écotoxicologiques des déchets n’entre pas dans le domaine d’application de cette Norme européenne.
La présente Norme européenne s’applique aux déchets solides et liquides.

Karakterizacija odpadkov - Priprava vzorcev odpadka za ekotoksikološke preskuse

General Information

Status
Withdrawn
Publication Date
30-Sep-2005
Withdrawal Date
13-Dec-2021
Technical Committee
Current Stage
9900 - Withdrawal (Adopted Project)
Start Date
03-Dec-2021
Due Date
26-Dec-2021
Completion Date
14-Dec-2021

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SLOVENSKI STANDARD
SIST EN 14735:2005
01-oktober-2005
Karakterizacija odpadkov - Priprava vzorcev odpadka za ekotoksikološke preskuse
Characterization of waste - Preparation of waste samples for ecotoxicity tests

Charakterisierung von Abfällen - Herstellung von Abfallproben für ökotoxikologische

Untersuchungen

Caractérisation des déchets - Caractérisation des déchets - Préparation des échantillons

de déchets en vue d'essais écotoxicologiques
Ta slovenski standard je istoveten z: EN 14735:2005
ICS:
13.030.01 Odpadki na splošno Wastes in general
SIST EN 14735:2005 en,fr,de

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

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SIST EN 14735:2005
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SIST EN 14735:2005
EUROPEAN STANDARD
EN 14735
NORME EUROPÉENNE
EUROPÄISCHE NORM
August 2005
ICS 13.030.01
English Version
Characterization of waste - Preparation of waste samples for
ecotoxicity tests

Caractérisation des déchets - Préparation des échantillons Charakterisierung von Abfällen - Herstellung von

de déchets en vue d'essais écotoxicologiques Abfallproben für ökotoxikologische Untersuchungen

This European Standard was approved by CEN on 27 June 2005.

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 Central Secretariat 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 Central Secretariat has the same status as the official

versions.

CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,

Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia,

Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels

© 2005 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 14735:2005: E

worldwide for CEN national Members.
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SIST EN 14735:2005
EN 14735:2005 (E)
Contents page

Foreword ..........................................................................................................................................................3

0 Introduction.........................................................................................................................................4

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

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

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

4 Equipment and reagents ....................................................................................................................7

5 Taking of laboratory sample ..............................................................................................................8

6 Transport .............................................................................................................................................8

7 Storage ................................................................................................................................................9

8 Waste characterization.....................................................................................................................10

9 Waste pre-treatment : particle size reduction (granular waste, monolithic waste,

paste-like waste and sludge)............................................................................................................10

10 Tests performed on terrestrial organisms ......................................................................................11

11 Tests performed on aquatic organisms ..........................................................................................14

12 Test report .........................................................................................................................................17

Annex A (normative) Preparation of test mixtures according to the ecotoxicity tests to be

performed ..........................................................................................................................................18

Annex B (informative) Ecotoxicity tests considered to establish this European Standard....................20

Bibliography...................................................................................................................................................44

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SIST EN 14735:2005
EN 14735:2005 (E)
Foreword

This European Standard (EN 14735:2005) has been prepared by Technical Committee CEN/TC 292

“Characterization of waste”, 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 February 2006, and conflicting national standards

shall be withdrawn at the latest by February 2006.

According to the CEN/CENELEC Internal Regulations, the national standards organizations of the

following countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Czech

Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy,

Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia, Slovenia,

Spain, Sweden, Switzerland and United Kingdom.
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SIST EN 14735:2005
EN 14735:2005 (E)
0 Introduction

Ecotoxicity can be estimated using two approaches: a chemical-specific approach and a toxicity-based

approach. Chemical analyses are compared, in the first case, to quality criteria or threshold values to

estimate ecotoxicity. In the second one, ecotoxicity is measured directly using biological tests. These

two approaches complement each other, however, determination of pollutants in complex mixtures of

unknown composition, that is a characteristic of many wastes, does not allow a relevant estimation of

ecotoxicity. For such samples, the toxicity based approach is usually recognised to be appropriate to

assess potential toxicity. Bioassays integrate, indeed, the effects of all contaminants including

additive, synergistic and antagonistic effects. They are sensitive to the bioavailable fraction of the

contaminants only. Finally, bioassays integrate the effects of all contaminants, including those, not

considered or detected by chemical analyses.

Ecotoxicity tests can be applied to wastes to identify their potential hazardous properties with respect

to the environment for classification purposes or to assess the risk related to a site-specific exposure

scenario.

0.1 Identification of properties potentially hazardous to the environment for classification

purposes

A classification system, based on the assessment of intrinsic properties, should be independent of an

exposure scenario. The main requirement, in order to establish a relevant system for classifying

wastes and for assessment of hazard properties, is to obtain comparable test results. This can only be

obtained if the ecotoxicity tests on wastes are carried out according to a unique procedure describing

more or less conventional test conditions (an exclusive dilution medium for terrestrial tests, a unique

L/S ratio for preparation of water extracts, a unique liquid / solid separation step etc). This procedure

should be applicable to a very wide range of waste materials whatever their physical properties are.

Any strategy for the assessment of properties potentially hazardous to the environment used in a

classification system should include test organisms representing the terrestrial and the aquatic

compartment. Both types of tests should be considered because they expand the range of effect

expression due to differences in species sensitivity and exposure. For this specific purpose, the water

extracts preparations for toxicity testing do not simulate leaching from wastes under environmental

conditions but measure the water available fraction of the toxic components of the wastes.

0.2 Site-specific exposure scenario

The second application of ecotoxicity tests to wastes refers to a risk assessment approach. In this

particular case, the test strategy should model site specific exposure conditions and should take into

account the transfer of contaminants via the food chain and to surface and ground water by run-off or

leaching. This application concerns firstly the definition of generic scenarios frequently encountered

(e.g. wastes deposit in stockpiles, re-use of wastes) and focus on the relevant way of exposure to

terrestrial and aquatic organisms.

This European Standard describes the necessary steps to be performed before carrying out

ecotoxicity tests on wastes within the context of assessment of ecotoxic properties used in a

classification system.
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SIST EN 14735:2005
EN 14735:2005 (E)
1 Scope

This European Standard describes the necessary steps to be performed before carrying out

ecotoxicity tests on wastes. The purpose of this European Standard 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 European Standard 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

European Standard.
This European Standard is applicable to solid and liquid wastes.
2 Normative references

The following referenced documents are indispensable for the application of this European Standard.

For dated references, only the edition cited applies. For undated references, the latest edition of the

referenced document (including any amendments) applies.

prEN 14899, Characterization of waste - Sampling of waste materials - Framework for the preparation

and application of a Sampling Plan

EN 12457-2:2002, Characterization of waste – Leaching – Compliance test for leaching of granular

waste materials and sludges – Part 2: One stage batch test at a liquid to solid ratio of 10 l/kg for

materials with particle size below 4 mm (without or with size reduction)

EN ISO 5667-3, Water quality - Sampling - Part 3: Guidance on the preservation and handling of water

samples (ISO 5667-3:2003)
ISO 10390, Soil quality – Determination of pH

ISO 11268-1, Soil quality – Effects of pollutants on earthworms (Eisenia fetida) – Part 1: Determination

of acute toxicity using artificial soil substrate

ISO 11465, Soil quality – Determination of dry matter and water content on a mass basis – Gravimetric

method

ISO 14238:1997, Soil quality – Biological methods – Determination of nitrogen mineralization and

nitrification in soils and the influence of chemicals on these processes
1) Definition of soil substrate.
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SIST EN 14735:2005
EN 14735:2005 (E)
3 Terms and definitions

For the purposes of this European Standard, the following terms and definitions apply.

3.1
dilution medium

liquid or solid used for the preparation of control vessels and the preparation of test mixture

3.2
ecotoxicological properties

potential adverse effects to biological systems which a waste has an inherent capacity to cause

3.3
eluate
solution recovered from a leaching test
[EN 12457-2:2002]
3.4
granular waste
waste not being monolithic, nor a liquid, a gas or a sludge
[EN 12457-2:2002]
3.5
laboratory sample
sample or subsample(s) sent to or received by the laboratory (IUPAC definition)

NOTE 1 When the laboratory sample is further prepared (reduced) by subdividing, mixing, grinding or by

combinations of these operations, the result is the test sample. When no preparation of the laboratory sample is

required, the laboratory sample is the test sample. A test portion is removed from the test sample for the

performance of the test or for analysis. The laboratory sample is the final sample from the point of view of sample

collection but it is the initial sample from the point of view of the laboratory.

NOTE 2 Several laboratory samples can be prepared and sent to different laboratories or to the same

laboratory for different purposes. When sent to the same laboratory, the set is generally considered as a single

laboratory sample and is documented as a single sample.
3.6
leachant
liquid used in a leaching test

NOTE For the purpose of this European Standard the leachant is water as specified in Clause 4.

3.7
leaching test

test during which a material is put into contact with a leachant and some constituents of the material

are extracted
3.8
liquid sludge

sludge that has the characteristic of a liquid as specified in the definition of a liquid waste

3.9
liquid waste

waste that completely flows out of a calibrated opening, down to the upper level of the opening within

a limited period of time (see Annex B of EN 12457-2:2002)
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SIST EN 14735:2005
EN 14735:2005 (E)
3.10
monolithic waste

material which has dimensional, physical and mechanical properties that comply with the criteria

defined in an on going document
3.11
paste-like material
material of soft plastic or wet cement consistency – usually smooth
3.12
sludge

mixture of liquid and solid separated from various types of liquid as a result of natural or artificial

processes
[EN 12457-2:2002]
3.13
test mixture
mixture of the test portion (waste or water extract) with the dilution medium
3.14
test portion

amount or volume of the test sample taken for measurement of ecotoxicological properties by

biological testing and/or other properties of interest, usually of known weight or volume (adapted from

IUPAC definition)

NOTE 1 The test portion can be taken from the laboratory sample directly if no preparation of sample is

required (e.g. with liquids), but usually it is taken from the prepared test sample.

NOTE 2 A unit or increment of proper homogeneity, size and fineness, needing no further preparation, can be

a test portion.
3.15
test sample

sample, prepared from the laboratory sample, from which test portions are removed for biological

testing or analysis (adapted from IUPAC definition)

NOTE The preparation of the test sample can include particle size reduction, preparation of water extract etc.

3.16
water extract

solution obtained from a leaching test, a liquid/liquid extraction and a liquid/solid separation

(centrifugation)
4 Equipment and reagents
Usual laboratory equipment and the following.
4.1 Sieving equipment with sieves of 4 mm square mesh.

NOTE Due to sieving, contamination of the sample may occur to an extent that affects the leaching of some

constituents of concern e.g. cobalt and tungsten from tungsten carbide equipment or chromium, nickel and

molybdenum from stainless steel equipment.
4.2 Crushing equipment: jaw crusher or cutting device.

NOTE Due to crushing, contamination of the sample may occur to an extent that affects the leaching of

some constituents of concern e.g. cobalt and tungsten from tungsten carbide equipment or chromium, nickel and

molybdenum from stainless steel equipment.
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SIST EN 14735:2005
EN 14735:2005 (E)
4.3 Balance of accuracy of at least 0,1 g.
4.4 End-over tumbler (5 rpm to 10 rpm) or rollertable rotating at about 10 rpm.

NOTE Other shaking or mixing devices can be used provided that they are proven to be equivalent.

4.5 Centrifuge operating at 2 500 g.

4.6 Filtering apparatus, either a vacuum filtration device (between 30 kPa and 70 kPa) (300 mbars

to 700 mbars) or a high pressure filtration apparatus (< 0,5 MPa) (5 bars).
4.7 Mixer.
4.8 pH meter.

4.9 Glass or high density polyethylene(HDPE)/polypropylene(PP) bottles in accordance with

EN ISO 5667-3, glass bottles having caps of inert material, for example polytetrafluoroethylene.

Rinsing is compulsory.

NOTE For inorganic constituents HDPE/PP bottles are preferred, except for samples tested for mercury.

4.10 Distilled water, demineralized water or deionized water with a conductivity < 5 µS/cm.

5 Taking of laboratory sample

Obtain a laboratory sample of a quantity sufficient for the number of tests to be performed and in

accordance with the requirements of biological standardised methods to be used.

The laboratory sample should be obtained according to the guide to the preparation of a sampling plan

for liquid and granular waste materials including paste-like materials and sludge, under development

by CEN/TC 292/WG 1.

Special precautions should be taken to avoid any contamination of laboratory samples by material of

sampling devices and/or storage equipment, according to prEN 14899.

NOTE Sampling devices are described in a standard under development by prCEN/TR 15310-2.

Some wastes are subject to chemical, physical and biological changes as soon as they are collected

(e.g. wastes that are fermentable, subject to oxidation or carbonation and wastes that contain volatile

substances). Possible changes shall be considered and sampling conditions shall be designed

accordingly in order to limit the effects of such changes on the results of ecotoxicity tests.

However, addition of preservatives (e.g. acids, basic solutions, biocides) in order to retard chemical

and biological activity is prohibited.
6 Transport

Transport of laboratory samples shall be performed in the dark, in tight containers fully filled with the

waste to be tested. However, special precautions should be taken for transport and storage of sludge

or other microbial active wastes. Containers can become pressurised due to gas production and

explosions may occur. For such laboratory samples, containers should not be completely filled.

Nevertheless, headspace shall not exceed 10 % of the total capacity of the container. Manual release

of pressure during and after transport may be necessary.

The container material shall be appropriate. The container, in which the waste laboratory sample is

transported, and the stopper shall not react with the constituents of the sample and shall not be a

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SIST EN 14735:2005
EN 14735:2005 (E)

cause of contamination. Wastes shall be stored in polyethylene, polypropylene, polytetrafluoroethene

(PTFE) or glass containers. However, security aspects shall be considered, including the risk of

explosion due to gas generation (for example glass vessels are not suitable for sludge samples).

Transport of waste laboratory samples should be as short as possible. Possible changes shall be

considered and transport conditions shall be designed accordingly in order to limit the effects of such

changes on the results of ecotoxicity tests. Transport time shall be regarded as part of storage time.

A transport time of less than 48 h and/or a low temperature conditions shall be (4 ± 2) °C in order to

appropriately to maintain the properties of laboratory samples.
7 Storage
7.1 General

Storage should be carried out in the containers defined in Clause 6. Possible changes shall be

considered and storage conditions shall be designed accordingly in order to limit the effects of such

changes on the results of ecotoxicity tests.
7.2 Waste sample

Storage time starting from reception of laboratory sample and ending with the start of definitive tests

should be as short as possible.

A storage time of less than two months and low temperature conditions shall be (4 ± 2) °C in order to

appropriately to maintain the properties of waste samples.
NOTE Freezing may induce changes of characteristics of the waste sample.
7.3 Water extracts

Water extracts should be stored at (4 ± 2) °C in polyethylene, polypropylene, polytetrafluoroethene

(PTFE) or glass containers. Before testing, the containers shall be filled with a headspace less than

5 %.
NOTE 1 Freezing may induce changes of characteristics of water extracts.

It is recommended to minimise the time between the start of the different tests to be performed on the

same laboratory sample in order to minimise its changes.

Ecotoxicological tests shall start immediately after production of water extract as specified in the

applicable standard for the considered ecotoxicity test and in no case later than 72 h after production

of water extract. If a range-finding test and a definitive test are performed, the definitive test shall be

completed within 10 days after production of the water extract.

For longer tests (e.g. semi-static chronic tests), several water extracts shall be produced and used

within 10 days after production.

If definitive test results are not in accordance with the range-finding test, water extraction shall be

repeated and the test shall be performed on the new water extract.

NOTE 2 It could be possible to extend the duration of storage if it has been proved that no modification of

toxicity occurred within the storage period (e.g. carry out the same ecotoxicity test immediately after extraction

period and at the end of storage period).
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SIST EN 14735:2005
EN 14735:2005 (E)
8 Waste characterization

The following characteristics shall be determined prior to the performance of ecotoxicity tests:

 pH, according to ISO 10390 for granular waste, monolithic waste, paste-like waste and sludge;

 dry matter content, according to ISO 11465 for granular waste, monolithic waste, paste-like waste

and sludge (see Clause 9);

 water holding capacity, according to Annex A of ISO 14238:1997 for granular waste, monolithic

waste, paste-like waste and sludge (see note).

NOTE The method described in Annex A of ISO 14238:1997 has been found to be appropriate for most of

the different kinds of waste.
9 Waste pre-treatment : particle size reduction (granular waste, monolithic
waste, paste-like waste and sludge)

Both ecotoxicological and leaching tests are performed on material which originally and after

pretreatment has a particle size less than 4 mm.

The tests shall be made on material with a grain size of at least 95 % (mass) less than 4 mm.

Therefore, the laboratory sample shall be sieved (4.1). If oversized material exceeds 5 % (mass) the

entire oversized fraction shall be crushed with crushing equipment (4.2). On no account shall the

material be finely ground. Non-crushable material (e.g. metallic parts such as nuts, bolts, scrap) in the

sample shall be separated and the weight and nature of the material shall be recorded. The method of

size-reduction applied shall be documented and recorded in the test report. Irrespective of any

necessary size reduction, the separate fractions with the exception of the non-crushable material shall

be mixed to constitute the test sample. If the laboratory sample cannot be crushed or sieved because

of its moisture content, it is allowed, only in this case, to dry the laboratory sample. The drying

temperature shall not exceed 40 °C.

NOTE 1 Fibrous materials, paste-like waste and plastics can often be size-reduced after cryogenic treatment.

The sample is usually plunged into liquid nitrogen (- 196 °C) just before crushing to make it fragile and brittle. It

also limits the overheating during crushing. As a result, the sample obtained is fine and perfectly homogeneous.

NOTE 2 In order to minimise the possible contamination during the sieving, fragmentation and splitting, it is

recommended, before preparing the test sample, to process a portion of the laboratory sample through the

devices for sieving, fragmentation and splitting and to discard such material thereafter. This recommendation

does not cover the situation described in the notes under 4.1 and 4.2.

For this European Standard, any other waste pre-treatment is excluded ; especially, the test sample

which shall not be further dried. The determination of the dry matter content ratio and the moisture

content ratio shall be determined on a dedicated test portion. The moisture content of the test sample

shall be determined at (105 ± 5) °C. It shall be taken into account when adjusting the L/S ratio in

leaching test. The dry mass of the sample shall be determined at (105 ± 5) °C according to ISO 11465

and the dry matter content ratio is calculated as follows:
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EN 14735:2005 (E)
DR = 100 × (1)
where
DR is the dry matter content ratio (%);
M is the mass of the dried test portion (kg);
M is the mass of undried test portion (kg).
The moisture content ratio is calculated as follows:
(M − M )
W D
MC = 100 × (2)
where
MC is the moisture content ratio (%).

NOTE 3 The basis for the calculation of the moisture content is the mass of the moisture content of the dry

residue in this European Standard, as specified in ISO 11465 (for the determination of the water content of soil). It

should be noted that in EN 12880 (for the determination of water content of sludge), the water content is

calculated on the basis of the raw mass.

NOTE 4 The above moisture content determination could be not accurate enough in some cases (e.g. large

amount of volatile or unstable compounds). In such cases a direct determination of the true water content should

be performed and the moisture content calculated accordingly.
10 Tests performed on terrestrial organisms
10.1 General considerations

The determination of ecotoxicological properties of wastes under conventional conditions requires

using a dilution medium as inert as possible. This dilution medium shall allow the survival and the

good development of organisms during the test period. Both requirements may be difficult to reconcile

particularly considering plant growth inhibition tests and microbial tests (tests that required indigenous

population of micro-organisms).

In order to fulfil theses requirements, the dilution medium called "artificial soil" shall be used unless

otherwise specified in the standardized terrestrial test methods. The same medium shall be used for

both control and dilution.

Moreover, preparation of a medium should be reproducible to allow comparison of ecotoxicity tests

results.

Several standardized ecotoxicity tests were considered to establish the following conditions for testing

wastes on terrestrial organisms. This compilation of tests is given in Annex B.

Preparation of test mixtures may differ according to the type of waste and according to the ecotoxicity

tests to be performed. Preparation of the different test mixtures is summarized in Annex A.

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EN 14735:2005 (E)
10.2 Dilution medium

The dilution medium shall have the following composition (such as defined in ISO 11268-1):

 sphagnum peat finely ground and with no visible plant remains: 10 % (percentage expressed on

dry mass basis);

 kaolinite clay containing not less than 30 % kaolinite: 20 % (percentage expressed on dry mass

basis);

 industrial quartz sand (more than 50 % of particle size from 0,05 mm to 0,20 mm): 70 %

(percentage expressed on dry mass basis).

Calcium carbonate (CaCO ), pulverised and of recognised analytical grade is added to bring the pH of

the wetted substrate to 6,0 ± 0,5 (generally between 0,5 % and 1 % of the mass of the dry ingredients).

Water (4.10) or mixture of water extract with water is added to the dilution medium to reach the

percentage of the total water holding capacity recommended for each test organism.

10.3 Introduction of waste into the dilution medium
10.3.1 General

Preparation of test mixtures differs according to the waste to be tested (see Annex A). The different

methods of preparation are described below.
10.3.2 Monolithic waste, granular waste, paste-like waste and sludge

Different methods can be applied to introduce the test portion into the dilution medium. Several

parameters influence the selection of introduction method such as physical properties of waste or

amounts to be tested. The following methods are recommended:

 for small amounts, introduce the test portion in the water (or in part of it) necessary to wet the

dilution medium, then mix this suspension thoroughly with the dilution medium;

 for large amounts, mix the test portion thoroughly with the already hydrated dilution medium;

 for hydrophobic waste, mix the test portion thoroughly with the dilution medium, then add the

water necessary
...

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