ISO 14442:2006
(Main)Water quality — Guidelines for algal growth inhibition tests with poorly soluble materials, volatile compounds, metals and waste water
Water quality — Guidelines for algal growth inhibition tests with poorly soluble materials, volatile compounds, metals and waste water
ISO 14442:2006 provides procedures, not covered by the methods described in ISO 8692 and ISO 10253, for testing difficult substances for inhibition of algal growth. The main subjects covered by ISO 14442:2006 are the methods for preparing the test substance for testing and the procedures needed to carry out an appropriate test. The following test substances are covered by this guideline: poorly soluble pure organic compounds; poorly soluble mixtures of organic substances; poorly soluble inorganic materials; volatile substances; waste waters and environmental samples containing water and sediments; coloured and/or turbid samples; compounds of heavy metals. The following methods of addition are covered: direct; dispersion; water-soluble and water-accommodated fractions. Some guidelines related to the analytical procedures and to the interpretation of the results have been included.
Qualité de l'eau — Lignes directrices pour essais d'inhibition de la croissance algale avec des matières peu solubles, des composés volatils, des métaux et des eaux résiduaires
L'ISO 14442:2006 fournit des modes opératoires, non couverts par les méthodes décrites dans l'ISO 8692 et l'ISO 10253, applicables aux essais d'inhibition de la croissance algale impliquant des substances difficiles. Les principaux sujets traités dans l'ISO 14442:2006 sont les méthodes de préparation de la substance d'essai ainsi que les modes opératoires nécessaires pour effectuer ces essais de façon appropriée. Ces lignes directrices sont applicables aux substances d'essai suivantes: les composés organiques purs peu solubles; les mélanges de substances organiques peu solubles; les matériaux inorganiques peu solubles; les substances volatiles; les eaux résiduaires et les échantillons environnementaux contenant de l'eau et des sédiments; les échantillons colorés et/ou turbides; les composés de métaux lourds. Les méthodes d'addition suivantes sont couvertes: directe; dispersion; fractions en suspension dans l'eau et fractions solubles dans l'eau. Certaines lignes directrices traitant des modes opératoires d'analyse et de l'interprétation des résultats sont également incluses.
Kakovost vode - Smernice za preskuse zaviranja rasti alg s slabo topnimi materiali, hlapnimi spojinami, kovinami in odpadno vodo
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-februar-2007
Kakovost vode - Smernice za preskuse zaviranja rasti alg s slabo topnimi
materiali, hlapnimi spojinami, kovinami in odpadno vodo
Water quality -- Guidelines for algal growth inhibition tests with poorly soluble materials,
volatile compounds, metals and waste water
Qualité de l'eau -- Lignes directrices pour essais d'inhibition de la croissance algale avec
des matières peu solubles, des composés volatils, des métaux et des eaux résiduaires
Ta slovenski standard je istoveten z: ISO 14442:2006
ICS:
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
INTERNATIONAL ISO
STANDARD 14442
Second edition
2006-04-01
Water quality — Guidelines for algal
growth inhibition tests with poorly
soluble materials, volatile compounds,
metals and waste water
Qualité de l'eau — Lignes directrices pour essais d'inhibition de la
croissance algale avec des matières peu solubles, des composés
volatils, des métaux et des eaux résiduaires
Reference number
©
ISO 2006
PDF disclaimer
This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but
shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
accepts no liability in this area.
Adobe is a trademark of Adobe Systems Incorporated.
Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation
parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In
the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
© ISO 2006
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 copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2006 – All rights reserved
Contents Page
Foreword. iv
1 Scope . 1
2 Normative references . 2
3 Analytical characterization of test materials and confirmation of concentrations and
stability . 2
4 Poorly soluble organic substances . 3
4.1 General. 3
4.2 Preparation of saturated and supersaturated solutions. 3
4.3 Solvent addition. 4
4.4 Dispersion using an emulsifying agent. 5
4.5 Interference with algal growth and its measurement. 5
5 Poorly soluble mixtures of organic substances. 6
5.1 General. 6
5.2 Preparation of test media. 6
5.3 Test performance. 7
6 Poorly soluble inorganic materials. 7
7 Volatile substances . 8
7.1 General. 8
7.2 Test system and growth medium. 8
7.3 Test performance. 9
7.4 Interference with algal growth. 9
8 Waste waters and environmental aqueous samples . 9
9 Coloured and/or turbid samples . 10
10 Metals and metal compounds . 11
10.1 Introduction . 11
10.2 Modification of algal growth inhibition test procedures for testing materials containing
heavy metals . 11
11 pH buffering. 12
12 Interpretation of the results . 13
Bibliography . 14
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 14442 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological
methods.
This second edition cancels and replaces the first edition (ISO 14442:1999), which has been technically
revised.
iv © ISO 2006 – All rights reserved
INTERNATIONAL STANDARD ISO 14442:2006(E)
Water quality — Guidelines for algal growth inhibition tests with
poorly soluble materials, volatile compounds, metals and waste
water
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This International Standard does not purport to address all of the safety problems, if any,
associated with its use. It is the responsibility of the user to establish appropriate safety and health
practices and to ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted according to this standard be carried
out by suitably trained staff.
1 Scope
This International Standard provides procedures, not covered by the methods described in ISO 8692 and
ISO 10253, for testing difficult substances for inhibition of algal growth.
The main subjects covered by the guideline are the methods for preparing the test substance for testing and
the procedures needed to carry out an appropriate test. The following test substances are covered by this
guideline:
a) poorly soluble pure organic compounds;
b) poorly soluble mixtures of organic substances;
c) poorly soluble inorganic materials;
d) volatile substances;
e) waste waters and environmental samples containing water and sediments;
f) coloured and/or turbid samples;
g) compounds of heavy metals.
The following methods of addition are covered:
⎯ direct;
⎯ dispersion;
⎯ water-soluble and water-accommodated fractions.
Some guidelines related to the analytical procedures and to the interpretation of the results have been
included.
References to documents describing the background for the testing of difficult substances are given in the
Bibliography.
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 5667-16, Water quality — Sampling — Part 16: Guidance on biotesting of samples
ISO 8692, Water quality — Freshwater algal growth inhibition test with unicellular green algae
ISO 10253, Water quality — Marine algal growth inhibition test with Skeletonema costatum and
Phaeodactylum tricornutum
3 Analytical characterization of test materials and confirmation of concentrations
and stability
Analytical characterization of test substances and materials and the confirmation of their concentrations and
stability in the testing environment is of major concern of regulatory authorities. Such activities are usually not
an integral part of this International Standard algal growth inhibition test methods.
However, there may be situations where analysis may assist in defining the appropriate exposure conditions
of test materials and chemicals and/or in the interpretation of the results.
The relevant properties of substances and materials can be assessed from basic properties such as solubility
in water, partition coefficient (lg P ), Henry's constant, photochemical and hydrolytic stability and
ow
biodegradability.
Analytical confirmation is strongly recommended in order to confirm test substance concentrations and is
required for the calculation of effective concentration (EC) values of volatile substances (Clause 7). If losses
due to adsorption on the test vessels or during transfer of test solutions and media occur, then analytical
confirmation are of particular importance. This aspect is also specified in ISO 5667-16.
Due to the batch test system used for algal growth inhibition tests, loss of substances due to biodegradation
(nearly all algal cultures contain bacteria), photodegradation, hydrolysis and/or adsorption cannot always be
avoided. A decrease in measured concentrations is difficult to prevent by technical means, and is therefore
considered acceptable for algal growth inhibition tests.
The following precautions are suggested for maintaining test substance concentrations in algal growth
inhibition tests:
a) sterilization of media and equipment to reduce the effect of bacterial growth;
b) change of the light quality to prevent photodegradation of test substances;
c) avoidance of contact of test substance with water prior to testing to reduce hydrolytic decomposition;
d) treatment of glassware (e.g. silanization); the effectiveness of such a treatment varies from one chemical
to the other;
e) pre-conditioning of the glassware, before addition of the test media, with the test substance at
concentrations to be used in the test.
The effect of such technical measures is, if relevant and if possible, monitored by chemical analysis.
Water, waste water and organic/inorganic solids/liquids may contain components that may modify the
composition of the algal growth medium (by precipitation of a limiting nutrient, complexation of essential
elements, addition of nutrients), and subsequently may cause effects on algal growth not related to toxic
2 © ISO 2006 – All rights reserved
components. If such problems occur, it may be advisable to determine the content of key components of the
test material. Some relevant components are: calcium, magnesium, sodium, potassium, sulfate, chloride,
ammonium, nitrate, phosphate, copper, cobalt, nickel, zinc, cadmium, organic matter [i.e. measured as
Chemical Oxygen Demand (COD) and/or Total Organic Carbon (TOC)].
If the material contains a high concentration of readily degradable organic material, the subsequent bact
...
INTERNATIONAL ISO
STANDARD 14442
Second edition
2006-04-01
Water quality — Guidelines for algal
growth inhibition tests with poorly
soluble materials, volatile compounds,
metals and waste water
Qualité de l'eau — Lignes directrices pour essais d'inhibition de la
croissance algale avec des matières peu solubles, des composés
volatils, des métaux et des eaux résiduaires
Reference number
©
ISO 2006
PDF disclaimer
This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but
shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
accepts no liability in this area.
Adobe is a trademark of Adobe Systems Incorporated.
Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation
parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In
the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
© ISO 2006
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 copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2006 – All rights reserved
Contents Page
Foreword. iv
1 Scope . 1
2 Normative references . 2
3 Analytical characterization of test materials and confirmation of concentrations and
stability . 2
4 Poorly soluble organic substances . 3
4.1 General. 3
4.2 Preparation of saturated and supersaturated solutions. 3
4.3 Solvent addition. 4
4.4 Dispersion using an emulsifying agent. 5
4.5 Interference with algal growth and its measurement. 5
5 Poorly soluble mixtures of organic substances. 6
5.1 General. 6
5.2 Preparation of test media. 6
5.3 Test performance. 7
6 Poorly soluble inorganic materials. 7
7 Volatile substances . 8
7.1 General. 8
7.2 Test system and growth medium. 8
7.3 Test performance. 9
7.4 Interference with algal growth. 9
8 Waste waters and environmental aqueous samples . 9
9 Coloured and/or turbid samples . 10
10 Metals and metal compounds . 11
10.1 Introduction . 11
10.2 Modification of algal growth inhibition test procedures for testing materials containing
heavy metals . 11
11 pH buffering. 12
12 Interpretation of the results . 13
Bibliography . 14
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 14442 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological
methods.
This second edition cancels and replaces the first edition (ISO 14442:1999), which has been technically
revised.
iv © ISO 2006 – All rights reserved
INTERNATIONAL STANDARD ISO 14442:2006(E)
Water quality — Guidelines for algal growth inhibition tests with
poorly soluble materials, volatile compounds, metals and waste
water
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This International Standard does not purport to address all of the safety problems, if any,
associated with its use. It is the responsibility of the user to establish appropriate safety and health
practices and to ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted according to this standard be carried
out by suitably trained staff.
1 Scope
This International Standard provides procedures, not covered by the methods described in ISO 8692 and
ISO 10253, for testing difficult substances for inhibition of algal growth.
The main subjects covered by the guideline are the methods for preparing the test substance for testing and
the procedures needed to carry out an appropriate test. The following test substances are covered by this
guideline:
a) poorly soluble pure organic compounds;
b) poorly soluble mixtures of organic substances;
c) poorly soluble inorganic materials;
d) volatile substances;
e) waste waters and environmental samples containing water and sediments;
f) coloured and/or turbid samples;
g) compounds of heavy metals.
The following methods of addition are covered:
⎯ direct;
⎯ dispersion;
⎯ water-soluble and water-accommodated fractions.
Some guidelines related to the analytical procedures and to the interpretation of the results have been
included.
References to documents describing the background for the testing of difficult substances are given in the
Bibliography.
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 5667-16, Water quality — Sampling — Part 16: Guidance on biotesting of samples
ISO 8692, Water quality — Freshwater algal growth inhibition test with unicellular green algae
ISO 10253, Water quality — Marine algal growth inhibition test with Skeletonema costatum and
Phaeodactylum tricornutum
3 Analytical characterization of test materials and confirmation of concentrations
and stability
Analytical characterization of test substances and materials and the confirmation of their concentrations and
stability in the testing environment is of major concern of regulatory authorities. Such activities are usually not
an integral part of this International Standard algal growth inhibition test methods.
However, there may be situations where analysis may assist in defining the appropriate exposure conditions
of test materials and chemicals and/or in the interpretation of the results.
The relevant properties of substances and materials can be assessed from basic properties such as solubility
in water, partition coefficient (lg P ), Henry's constant, photochemical and hydrolytic stability and
ow
biodegradability.
Analytical confirmation is strongly recommended in order to confirm test substance concentrations and is
required for the calculation of effective concentration (EC) values of volatile substances (Clause 7). If losses
due to adsorption on the test vessels or during transfer of test solutions and media occur, then analytical
confirmation are of particular importance. This aspect is also specified in ISO 5667-16.
Due to the batch test system used for algal growth inhibition tests, loss of substances due to biodegradation
(nearly all algal cultures contain bacteria), photodegradation, hydrolysis and/or adsorption cannot always be
avoided. A decrease in measured concentrations is difficult to prevent by technical means, and is therefore
considered acceptable for algal growth inhibition tests.
The following precautions are suggested for maintaining test substance concentrations in algal growth
inhibition tests:
a) sterilization of media and equipment to reduce the effect of bacterial growth;
b) change of the light quality to prevent photodegradation of test substances;
c) avoidance of contact of test substance with water prior to testing to reduce hydrolytic decomposition;
d) treatment of glassware (e.g. silanization); the effectiveness of such a treatment varies from one chemical
to the other;
e) pre-conditioning of the glassware, before addition of the test media, with the test substance at
concentrations to be used in the test.
The effect of such technical measures is, if relevant and if possible, monitored by chemical analysis.
Water, waste water and organic/inorganic solids/liquids may contain components that may modify the
composition of the algal growth medium (by precipitation of a limiting nutrient, complexation of essential
elements, addition of nutrients), and subsequently may cause effects on algal growth not related to toxic
2 © ISO 2006 – All rights reserved
components. If such problems occur, it may be advisable to determine the content of key components of the
test material. Some relevant components are: calcium, magnesium, sodium, potassium, sulfate, chloride,
ammonium, nitrate, phosphate, copper, cobalt, nickel, zinc, cadmium, organic matter [i.e. measured as
Chemical Oxygen Demand (COD) and/or Total Organic Carbon (TOC)].
If the material contains a high concentration of readily degradable organic material, the subsequent bacterial
growth may disturb the algal growth measurement. When untreated (not filtered or centrifuged) waste water is
tested, contamination with other algal species may occur.
4 Poorly soluble organic substances
4.1 General
A pure substance is a substance with one major component containing minor components as impurities.
Poorly soluble substances are those with solubility limits below 100 mg/l in water. If, however, growth
inhibition occurs at concentrations much lower than the solubility limits in water or algal growth medium (the
limit in the medium may be different), then the poorly soluble substance can be tested as a water soluble
substance (added via a stock solution in test medium). This approach is usually not applicable to substances
with a water solubility below 1 mg/l to 10 mg/l (substances with a very low solubility).
The methods described in this clause therefore refer to testing of substances causing effects on algal growth
at concentrations at or around the solubility limit in water and to very low solubility substances.
Testing of no
...
NORME ISO
INTERNATIONALE 14442
Deuxième édition
2006-04-01
Qualité de l'eau — Lignes directrices
pour essais d'inhibition de la croissance
algale avec des matières peu solubles,
des composés volatils, des métaux et des
eaux résiduaires
Water quality — Guidelines for algal growth inhibition tests with poorly
soluble materials, volatile compounds, metals and waste water
Numéro de référence
©
ISO 2006
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Web www.iso.org
Publié en Suisse
ii © ISO 2006 – Tous droits réservés
Sommaire Page
Avant-propos. iv
1 Domaine d'application. 1
2 Références normatives . 2
3 Caractérisation analytique des matériaux d'essai et confirmation des concentrations et
de la stabilité . 2
4 Substances organiques peu solubles . 3
4.1 Généralités . 3
4.2 Préparation de solutions saturées et sursaturées . 4
4.3 Addition de solvants. 4
4.4 Dispersion à l'aide d'un agent émulsifiant . 5
4.5 Interférence avec la croissance algale et son mesurage . 6
5 Mélanges peu solubles de substances organiques. 7
5.1 Généralités . 7
5.2 Préparation des milieux d'essai . 7
5.3 Réalisation de l'essai. 8
6 Matériaux solides inorganiques peu solubles. 8
7 Substances volatiles . 9
7.1 Généralités . 9
7.2 Système d'essai et milieu de culture . 10
7.3 Rendement de l'essai . 10
7.4 Interférences avec la croissance algale . 10
8 Eaux résiduaires et échantillons environnementaux aqueux. 11
9 Échantillons colorés et/ou turbides. 12
10 Métaux et composés métalliques. 12
10.1 Introduction . 12
10.2 Modification des modes opératoires d'essai d'inhibition de la croissance algale pour
l'essai de matériaux contenant des métaux lourds. 13
11 Tamponnage du pH . 14
12 Interprétation des résultats . 15
Bibliographie . 16
Avant-propos
L'ISO (Organisation internationale de normalisation) est une fédération mondiale d'organismes nationaux de
normalisation (comités membres de l'ISO). L'élaboration des Normes internationales est en général confiée
aux comités techniques de l'ISO. Chaque comité membre intéressé par une étude a le droit de faire partie du
comité technique créé à cet effet. Les organisations internationales, gouvernementales et non
gouvernementales, en liaison avec l'ISO participent également aux travaux. L'ISO collabore étroitement avec
la Commission électrotechnique internationale (CEI) en ce qui concerne la normalisation électrotechnique.
Les Normes internationales sont rédigées conformément aux règles données dans les Directives ISO/CEI,
Partie 2.
La tâche principale des comités techniques est d'élaborer les Normes internationales. Les projets de Normes
internationales adoptés par les comités techniques sont soumis aux comités membres pour vote. Leur
publication comme Normes internationales requiert l'approbation de 75 % au moins des comités membres
votants.
L'attention est appelée sur le fait que certains des éléments du présent document peuvent faire l'objet de
droits de propriété intellectuelle ou de droits analogues. L'ISO ne saurait être tenue pour responsable de ne
pas avoir identifié de tels droits de propriété et averti de leur existence.
L'ISO 14442 a été élaborée par le comité technique ISO/TC 147, Qualité de l'eau, sous-comité SC 5,
Méthodes biologiques.
Cette deuxième édition annule et remplace la première édition (ISO 14442:1999), qui a fait l'objet d'une
révision technique.
iv © ISO 2006 – Tous droits réservés
NORME INTERNATIONALE ISO 14442:2006(F)
Qualité de l'eau — Lignes directrices pour essais d'inhibition de
la croissance algale avec des matières peu solubles, des
composés volatils, des métaux et des eaux résiduaires
AVERTISSEMENT — Il convient que l'utilisateur de la présente Norme internationale connaisse bien
les pratiques courantes de laboratoire. La présente Norme internationale n'a pas pour but de traiter
tous les problèmes de sécurité qui sont, le cas échéant, liés à son utilisation. Il incombe à l'utilisateur
de mettre en place des pratiques appropriées en matière d'hygiène et de sécurité, et de s'assurer de la
conformité à la réglementation nationale en vigueur.
IMPORTANT — Il est absolument essentiel que les essais conduits conformément à la présente norme
soient effectués par un personnel ayant reçu une formation adéquate.
1 Domaine d'application
La présente Norme internationale a pour objectif de fournir des modes opératoires, non couverts par les
méthodes décrites dans l'ISO 8692 et l'ISO 10253, applicables aux essais d'inhibition de la croissance algale
impliquant des substances difficiles.
Les principaux sujets traités dans ces lignes directrices sont les méthodes de préparation de la substance
d'essai ainsi que les modes opératoires nécessaires pour effectuer ces essais de façon appropriée. Ces
lignes directrices sont applicables aux substances d'essai suivantes:
a) composés organiques purs peu solubles;
b) mélanges de substances organiques peu solubles;
c) matériaux inorganiques peu solubles;
d) substances volatiles;
e) eaux résiduaires et échantillons environnementaux contenant de l'eau et des sédiments;
f) échantillons colorés et/ou turbides;
g) composés de métaux lourds.
Les méthodes d'addition suivantes sont couvertes:
⎯ directe;
⎯ dispersion;
⎯ fractions en suspension dans l'eau et fractions solubles dans l'eau.
Certaines lignes directrices traitant des modes opératoires d'analyse et de l'interprétation des résultats sont
également incluses.
Les références faites à des documents décrivant le contexte relatif aux essais sur des substances difficiles
sont récapitulées dans la Bibliographie.
2 Références normatives
Les documents de référence suivants sont indispensables pour l'application du présent document. Pour les
références datées, seule l'édition citée s'applique. Pour les références non datées, la dernière édition du
document de référence s'applique (y compris les éventuels amendements).
ISO 5667-16, Qualité de l'eau — Échantillonnage — Partie 16: Lignes directrices pour les essais biologiques
des échantillons
ISO 8692, Qualité de l'eau — Essai d'inhibition de la croissance des algues d'eau douce avec des algues
vertes unicellulaires
ISO 10253, Qualité de l'eau — Essai d'inhibition de la croissance des algues marines avec Skeletonema
costatum et Phaeodactylum tricornutum
3 Caractérisation analytique des matériaux d'essai et confirmation des
concentrations et de la stabilité
La caractérisation analytique des substances et des matériaux soumis à essai ainsi que la confirmation de
leur concentration et de leur stabilité dans l'environnement d'essai est une préoccupation majeure pour les
autorités réglementaires. Ces activités ne font normalement pas partie intégrante des méthodes d'essai
d'inhibition de la croissance algale développées dans la présente Norme internationale.
Cependant, il peut exister des situations dans lesquelles l'analyse peut contribuer à mieux définir les
conditions d'exposition des matériaux et des substances chimiques d'essai et/ou à faciliter l'interprétation des
résultats.
Les caractéristiques importantes des substances et matériaux peuvent être évaluées à partir des
caractéristiques de base telles que la solubilité dans l'eau, le coefficient de partage (lg P ), la constante de
ow
Henry, la stabilité photochimique et hydrolytique ainsi que la biodégradabilité.
Il est fortement conseillé de procéder à une analyse des concentrations de la substance chimique d'essai,
concentrations qui sont exigées pour le calcul des valeurs de concentration effective (CE) des substances
volatiles (Article 7). Si des pertes de substance ont lieu par adsorption sur les parois des récipients d'essai ou
pendant le transfert des solutions et du milieu, cette confirmation analytique sera essentielle. Cet aspect est
également spécifié dans l'ISO 5667-16.
En raison du système d'essai sous forme de lots utilisé pour les essais d'inhibition de la croissance algale, il
n'est pas toujours possible d'éviter les pertes de substance dues à la biodégradation (la plupart des cultures
algales contiennent des bactéries), la photodégradation, l'hydrolyse et/ou l'adsorption. La diminution des
concentrations mesurées étant difficile à empêcher par des moyens techniques, elle est considérée comme
acceptable dans le cadre des essais d'inhibition de la croissance algale.
Dans le cadre d'essais d'inhibition de la croissance algale, il est suggéré de prendre les précautions suivantes
pour maintenir à un niveau constant les concentrations de la substance d'essai:
a) stérilisation des milieux de culture et des matériels utilisés pour réduire les effets de la croissance
bactérienne;
b) changement de la qualité de la lumière pour empêcher une photodégradation des substances d'essai;
c) absence de tout contact de la substance d'essai avec l'eau avant l'essai pour réduire les risques liés à la
décomposition par hydrolyse;
d) traitement de la verrerie (par exemple par silanisation); ce type de traitement est plus ou moins efficace
selon la substance chimique concernée;
2 © ISO 2006 – Tous droits réservés
e) préconditionnement de la verrerie avec la substance d'essai aux concentrations utilisées au cours de
l'essai avant l'ajout du milieu d'essai.
Si nécessaire et si possible, contrôler par des analyses chimiques l'effet de ces mesures techniques.
L'eau, les eaux résiduaires, les liquides/solides organiques/inorganiques peuvent contenir des composés
susceptibles de modifier la composition du milieu de culture des algues (par précipitation d'un nutriment
limitant, complexation d'éléments essentiels, addition de nutriments) et par voie de conséquence, d'affecter de
façon non négligeable la croissance algale sans implication de composés toxiques. En pr
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