ISO/TS 20787:2017
(Main)Nanotechnologies - Aquatic toxicity assessment of manufactured nanomaterials in saltwater lakes using Artemia sp. Nauplii
Nanotechnologies - Aquatic toxicity assessment of manufactured nanomaterials in saltwater lakes using Artemia sp. Nauplii
ISO/TS 20787:2017 specifies a test method, aiming to maximize repeatability and reliability of testing, to determine whether MNMs are toxic to aquatic organisms, specifically Artemia sp. nauplius. ISO/TS 20787:2017 is intended to be used by ecotoxicological laboratories that are capable in the hatching and culturing of Artemia sp. and the evaluation of toxicity of nanomaterials using Artemia sp. nauplius. This method uses Artemia sp. nauplii in a simulated environment, artificial seawater, to assess effects of nanomaterials. ISO/TS 20787:2017 is applicable to MNMs that consist of nano-objects such as nanoparticles, nanopowders, nanofibres, nanotubes, nanowires, as well as aggregates and agglomerates of such MNMs.
Nanotechnologies - Evaluation de la toxicité des nanomatériaux manufacturés dans les lacs d’eau salés par des Artemia sp. Nauplii
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TECHNICAL ISO/TS
SPECIFICATION 20787
First edition
2017-11
Nanotechnologies - Aquatic toxicity
assessment of manufactured
nanomaterials in saltwater lakes using
Artemia sp. Nauplii
Nanotechnologies - Evaluation de la toxicité des nanomatériaux en
milieu aquatique par des Artemia sp
Reference number
©
ISO 2017
© ISO 2017, Published in Switzerland
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of
the requester.
ISO copyright office
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2017 – All rights reserved
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Materials . 5
4.1 Test organism . 5
4.2 Chemicals . 5
5 Technical equipment . 5
6 Preparation and characterization of dispersion of nanomaterial .6
6.1 Dispersion preparation . 6
6.2 Dispersion characterization . 7
6.3 Dispersion stability in stock suspension . 7
6.4 Dispersion stability in artificial seawater . 7
6.5 Preparation of exposure media for toxicity tests . 7
7 Hatching procedure . 7
7.1 General . 7
7.2 Dilution water . 7
7.3 Storage of cysts . 8
7.4 Disinfection of Artemia sp. cysts . 8
7.5 Hatching method of Artemia sp. cysts . 8
7.6 Harvesting of nauplii . 8
7.7 Calculation of hatching percentage . 9
8 Effect of nanomaterial on Artemia sp. nauplii . 9
8.1 Test groups and controls . 9
8.2 Test concentrations . 9
8.3 Exposure condition .10
8.4 Duration .10
8.5 Observations .10
8.6 Analytical measurements .10
9 Data analysis .10
10 Test report .10
10.1 Test procedure .10
10.2 Information to include in the report .11
10.2.1 Test nanomaterial .11
10.2.2 Test species .11
10.2.3 Test conditions.11
10.2.4 Bioassay results .11
11 Results validity .11
Bibliography .13
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.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).
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. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to the
World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following
URL: www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 229, Nanotechnologies.
iv © ISO 2017 – All rights reserved
Introduction
With the increasing development and use of manufactured nanomaterials (MNMs) in consumer and
other products, concern about the possible impact of MNMs on human and environmental health is
increasing. Various aquatic organisms (such as fish, daphnia, algae, etc.) are currently used to predict
the possible adverse effects of chemicals, including nanomaterials, on the aquatic environment. Brine
[42]
shrimp (Artemia sp.) are found nearly worldwide in saline lakes and pools, and are one of the most
widespread euryhaline organisms that are suitable for ecotoxicity testing. Artemia sp. nauplii can
be used to assess the effects of nanomaterials in salt water ecosystems, primarily salt lakes. Artemia
sp. usually live in salt lakes, and are almost never found in an open sea. This species also adapts to a
wide range of salinities (5 g/L to 300 g/L) and temperatures (6 °C to 40 °C). In fact, the physiologically
optimal levels of salinity for Artemia sp. are about 30 g/L to 35 g/L. Due to predators at these salt
levels, however, Artemia sp. seldom occur in natural habitats at salinities of less than 45 g/L to 80 g/L.
Favoured for the absence of predators and food competitors in such places, Artemia sp. develop very
dense populations.
There are several advantages to using Artemia sp. as a biological model in salt water aquatic toxicology:
a) Less concern about animal welfare than for a vertebrate species;
b) There is good knowledge of Artemia sp. biology and ecology;
c) Artemia sp. have a wide geographic distribution in salt water lakes and pools;
d) Tests performed on Artemia sp. nauplii are simple and cost-effective;
e) Small body size allows accommodation of Artemia sp. nauplii in small beakers or plates;
f) Artemia sp. adapt to a wide range of water salinity and temperature;
g) Artemia sp. are simple to maintain in the laboratory;
h) The life cycle of Artemia sp. is short, so it is suitable for growth, reproduction and short-term
toxicity tests;
i) Artemia sp. cysts are commercially and readily available so that the tests can be carried out
worldwide. The cysts can be stored for years under cool and dry conditions without losing viability.
Upon immersion in sea water, the free swimming nauplii will hatch within approximately 24 h;
j) Hatching from cysts gives organisms of similar age, genotype and physiological condition.
In recent years, several researchers around the world have used Artemia sp. as a test organism in aquatic
nanotoxicology (see References [1] to [35]). The lack of a standardized protocol for testing Artemia sp.
for aquatic toxicity means that data from these studies are more likely to be non-repeatable and non-
[22]
reliable. The goal of this document is to provide a standard protocol intended to generate reliable
aquatic toxicity data by testing Artemia sp., which can be used for ecotoxicity evaluation of MNMs in salt
water lake ecosystems.
TECHNICAL SPECIFICATION ISO/TS 20787:2017(E)
Nanotechnologies - Aquatic toxicity assessment of
manufactured nanomaterials in saltwater lakes using
Artemia sp. Nauplii
1 Scope
This document specifies a test method, aiming to maximize repeatability and reliability of testing, to
determine whether MNMs are toxic to aquatic organisms, specifically Artemia sp. nauplius.
This document is intended to be used by ecotoxicological laboratories that are capable in the hatching
and culturing of Artemia sp. and the evaluation of toxicity of nanomaterials using Artemia sp. nauplius.
This method uses Artemia sp. nauplii in a simula
...
TECHNICAL ISO/TS
SPECIFICATION 20787
First edition
2017-11
Nanotechnologies - Aquatic toxicity
assessment of manufactured
nanomaterials in saltwater lakes using
Artemia sp. Nauplii
Nanotechnologies - Evaluation de la toxicité des nanomatériaux en
milieu aquatique par des Artemia sp
Reference number
©
ISO 2017
© ISO 2017, Published in Switzerland
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of
the requester.
ISO copyright office
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2017 – All rights reserved
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Materials . 5
4.1 Test organism . 5
4.2 Chemicals . 5
5 Technical equipment . 5
6 Preparation and characterization of dispersion of nanomaterial .6
6.1 Dispersion preparation . 6
6.2 Dispersion characterization . 7
6.3 Dispersion stability in stock suspension . 7
6.4 Dispersion stability in artificial seawater . 7
6.5 Preparation of exposure media for toxicity tests . 7
7 Hatching procedure . 7
7.1 General . 7
7.2 Dilution water . 7
7.3 Storage of cysts . 8
7.4 Disinfection of Artemia sp. cysts . 8
7.5 Hatching method of Artemia sp. cysts . 8
7.6 Harvesting of nauplii . 8
7.7 Calculation of hatching percentage . 9
8 Effect of nanomaterial on Artemia sp. nauplii . 9
8.1 Test groups and controls . 9
8.2 Test concentrations . 9
8.3 Exposure condition .10
8.4 Duration .10
8.5 Observations .10
8.6 Analytical measurements .10
9 Data analysis .10
10 Test report .10
10.1 Test procedure .10
10.2 Information to include in the report .11
10.2.1 Test nanomaterial .11
10.2.2 Test species .11
10.2.3 Test conditions.11
10.2.4 Bioassay results .11
11 Results validity .11
Bibliography .13
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.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).
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. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to the
World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following
URL: www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 229, Nanotechnologies.
iv © ISO 2017 – All rights reserved
Introduction
With the increasing development and use of manufactured nanomaterials (MNMs) in consumer and
other products, concern about the possible impact of MNMs on human and environmental health is
increasing. Various aquatic organisms (such as fish, daphnia, algae, etc.) are currently used to predict
the possible adverse effects of chemicals, including nanomaterials, on the aquatic environment. Brine
[42]
shrimp (Artemia sp.) are found nearly worldwide in saline lakes and pools, and are one of the most
widespread euryhaline organisms that are suitable for ecotoxicity testing. Artemia sp. nauplii can
be used to assess the effects of nanomaterials in salt water ecosystems, primarily salt lakes. Artemia
sp. usually live in salt lakes, and are almost never found in an open sea. This species also adapts to a
wide range of salinities (5 g/L to 300 g/L) and temperatures (6 °C to 40 °C). In fact, the physiologically
optimal levels of salinity for Artemia sp. are about 30 g/L to 35 g/L. Due to predators at these salt
levels, however, Artemia sp. seldom occur in natural habitats at salinities of less than 45 g/L to 80 g/L.
Favoured for the absence of predators and food competitors in such places, Artemia sp. develop very
dense populations.
There are several advantages to using Artemia sp. as a biological model in salt water aquatic toxicology:
a) Less concern about animal welfare than for a vertebrate species;
b) There is good knowledge of Artemia sp. biology and ecology;
c) Artemia sp. have a wide geographic distribution in salt water lakes and pools;
d) Tests performed on Artemia sp. nauplii are simple and cost-effective;
e) Small body size allows accommodation of Artemia sp. nauplii in small beakers or plates;
f) Artemia sp. adapt to a wide range of water salinity and temperature;
g) Artemia sp. are simple to maintain in the laboratory;
h) The life cycle of Artemia sp. is short, so it is suitable for growth, reproduction and short-term
toxicity tests;
i) Artemia sp. cysts are commercially and readily available so that the tests can be carried out
worldwide. The cysts can be stored for years under cool and dry conditions without losing viability.
Upon immersion in sea water, the free swimming nauplii will hatch within approximately 24 h;
j) Hatching from cysts gives organisms of similar age, genotype and physiological condition.
In recent years, several researchers around the world have used Artemia sp. as a test organism in aquatic
nanotoxicology (see References [1] to [35]). The lack of a standardized protocol for testing Artemia sp.
for aquatic toxicity means that data from these studies are more likely to be non-repeatable and non-
[22]
reliable. The goal of this document is to provide a standard protocol intended to generate reliable
aquatic toxicity data by testing Artemia sp., which can be used for ecotoxicity evaluation of MNMs in salt
water lake ecosystems.
TECHNICAL SPECIFICATION ISO/TS 20787:2017(E)
Nanotechnologies - Aquatic toxicity assessment of
manufactured nanomaterials in saltwater lakes using
Artemia sp. Nauplii
1 Scope
This document specifies a test method, aiming to maximize repeatability and reliability of testing, to
determine whether MNMs are toxic to aquatic organisms, specifically Artemia sp. nauplius.
This document is intended to be used by ecotoxicological laboratories that are capable in the hatching
and culturing of Artemia sp. and the evaluation of toxicity of nanomaterials using Artemia sp. nauplius.
This method uses Artemia sp. nauplii in a simula
...
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