EN ISO 10808:2010

Nanotechnologies - Characterization of nanoparticles in inhalation exposure chambers for inhalation toxicity testing (ISO 10808:2010)

ISO 10808:2010 specifies requirements for, and gives guidance on, the characterization of airborne nanoparticles in inhalation exposure chambers for the purpose of inhalation toxicity studies in terms of particle mass, size distribution, number concentration and composition.

Nanotechnologien - Charakterisierung von Nanopartikeln in Inhalationskammern zur Prüfung auf Toxizität nach Inhalation (ISO 10808:2010)

Diese Internationale Norm legt Anforderungen an die für Inhalationstoxizitätsuntersuchungen dienende Charakterisierung von luftgetragenen Nanopartikeln in Inhalationskammern hinsichtlich Partikelmasse, -größenverteilung, -anzahlkonzentration und -zusammensetzung fest und liefert Hilfestellung dafür.

Nanotechnologies - Caractérisation des nanoparticules dans les chambres d'inhalation par exposition pour les essais de toxicité par inhalation (ISO 10808:2010)

L'ISO 10808:2010 spécifie les exigences et donne les directives en termes de caractérisation des nanoparticules en suspension dans des chambres d'exposition par inhalation pour des études de toxicité par inhalation orientées vers la concentration massique, la distribution de taille, la concentration en nombre et la composition des particules.

Nanotehnologija - Karakterizacija nanodelcev v inhalacijskih komorah za preskušanje toksičnosti pri vdihovanju (ISO 10808:2010)

Ta mednarodni standard določa zahteve in svetuje v zvezi s karakterizacijo nanodelcev v zraku v inhalacijskih komorah za preučevanje toksičnosti pri vdihovanju, kar zadeva maso delcev, porazdelitev velikosti, koncentracijo števila in sestavo.

General Information

• Status: Published
• Publication Date: 14-Dec-2010
• Withdrawal Date: 29-Jun-2011
• ICS: 07.030 - Physics. Chemistry
• ICS: 07.120 - Nanotechnologies
• ICS: 71.100.01 - Products of the chemical industry in general
• Technical Committee: CEN/TC 352 - Nanotechnologies
• Drafting Committee: CEN/TC 352 - Nanotechnologies
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 15-Dec-2010
• Completion Date: 15-Dec-2010
• Ref Project: SIST EN ISO 10808:2011 - Nanotechnologies - Characterization of nanoparticles in inhalation exposure chambers for inhalation toxicity testing (ISO 10808:2010)

Overview

EN ISO 10808:2010 (ISO 10808:2010) - Nanotechnologies - provides requirements and guidance for the characterization of airborne nanoparticles in inhalation exposure chambers used in inhalation toxicity testing. The standard focuses on monitoring and reporting particle mass, particle size distribution, particle number concentration and composition to support reliable inhalation toxicology studies and exposure assessment for nanomaterials.

Key topics and requirements

• Scope and purpose
  • Specifies how to characterize nanoparticles in inhalation exposure chambers (nose-only, whole-body) for toxicity testing.
• Particle measurement systems
  • Recommends use of systems such as Differential Mobility Analyzers (DEMC/DEMS), Condensation Particle Counters (CPC) and integrated DMAS for number‑based size distribution and concentration measurement.
  • Advises morphological and compositional analysis using TEM/SEM with EDXA.
• Measured parameters
  • Number concentration, number-based size distribution (e.g., CMD, GMD, GSD), mass concentration, surface area (where relevant) and chemical composition.
• Monitoring locations
  • Emphasizes sampling from the animal’s breathing zone to ensure representative exposure data.
• Data assessment and reporting
  • Defines how to calculate and report geometric mean diameter (GMD), geometric standard deviation (GSD) and other distribution statistics.
  • Includes requirements for test reports to document chamber performance and nanoparticle characterization.
• References and harmonization
  • Normative references include ISO 15900, ISO/TS 27687 and relevant OECD inhalation test guidelines (TG 403, 412, 413).

Applications and users

Who uses EN ISO 10808:2010:

• Inhalation toxicologists and academic researchers conducting nano‑inhalation studies.
• Contract research organizations (CROs) and laboratories running animal inhalation exposure chambers.
• Regulatory bodies and safety assessors evaluating inhalation risk of nanomaterials.
• Occupational hygienists and industrial R&D teams designing exposure controls and risk mitigation.
• Instrument manufacturers and QA personnel validating chamber monitoring systems.

Practical uses:

• Designing and validating inhalation exposure studies for nanoparticles (e.g., TiO2, silica, carbon-based).
• Generating standardized exposure metrics for regulatory submissions, risk assessments and reproducible research.
• Ensuring representative sampling (breathing zone) and multi-parameter characterization (number, size, mass, composition).

Related standards

• ISO 15900 - Differential electrical mobility analysis for aerosol particles
• ISO/TS 27687 - Nanotechnologies terminology and definitions
• OECD TG 403, 412, 413 and GD 39 - Inhalation toxicity guidance

Keywords: EN ISO 10808:2010, ISO 10808:2010, nanotechnologies, nanoparticles, inhalation exposure chambers, inhalation toxicity testing, DMAS, particle size distribution, number concentration, TEM, SEM.

Standards Content (Sample)

SLOVENSKI STANDARD
01-november-2011
1DQRWHKQRORJLMD.DUDNWHUL]DFLMDQDQRGHOFHYYLQKDODFLMVNLKNRPRUDK]D
SUHVNXãDQMHWRNVLþQRVWLSULYGLKRYDQMX,62
Nanotechnologies - Characterization of nanoparticles in inhalation exposure chambers
for inhalation toxicity testing (ISO 10808:2010)
Nanotechnologien - Charakterisierung von Nanopartikeln in Inhalationskammern zur
Prüfung auf Toxizität nach Inhalation (ISO 10808:2010)
Nanotechnologies - Caractérisation des nanoparticules dans les chambres d'inhalation
par exposition pour les essais de toxicité par inhalation (ISO 10808:2010)
Ta slovenski standard je istoveten z: EN ISO 10808:2010
ICS:
07.120 Nanotehnologije Nanotechnologies
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN ISO 10808
NORME EUROPÉENNE
EUROPÄISCHE NORM
December 2010
ICS 07.030
English Version
Nanotechnologies - Characterization of nanoparticles in
inhalation exposure chambers for inhalation toxicity testing (ISO
10808:2010)
Nanotechnologies - Caractérisation des nanoparticules Nanotechnologien - Charakterisierung von Nanopartikeln in
dans les chambres d'inhalation par exposition pour les Inhalationskammern zur Prüfung auf Toxizität nach
essais de toxicité par inhalation (ISO 10808:2010) Inhalation (ISO 10808:2010)
This European Standard was approved by CEN on 10 December 2010.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same
status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.

EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2010 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 10808:2010: E
worldwide for CEN national Members.

Contents Page
Foreword .3

Foreword
This document (EN ISO 10808:2010) has been prepared by Technical Committee ISO/TC 229
“Nanotechnologies” in collaboration with Technical Committee CEN/TC 352 “Nanotechnologies” the
secretariat of which is held by BSI.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by June 2011, and conflicting national standards shall be withdrawn at
the latest by June 2011.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: 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, Romania, Slovakia, Slovenia, Spain,
Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 10808:2010 has been approved by CEN as a EN ISO 10808:2010 without any modification.

INTERNATIONAL ISO
STANDARD 10808
First edition
2010-12-15
Nanotechnologies — Characterization of
nanoparticles in inhalation exposure
chambers for inhalation toxicity testing
Nanotechnologies — Caractérisation des nanoparticules dans les
chambres d'inhalation par exposition pour les essais de toxicité par
inhalation
Reference number
ISO 10808:2010(E)
©
ISO 2010
ISO 10808:2010(E)
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Published in Switzerland
ii © ISO 2010 – All rights reserved

ISO 10808:2010(E)
Contents Page
Foreword .iv
Introduction.v
1 Scope.1
2 Normative references.1
3 Terms and definitions .1
3.1 Particle measuring systems.2
4 Test substance monitoring method .4
4.1 Principle .4
4.1.1 Exposure .4
4.1.2 Particle properties.4
4.2 Preparation of system.4
4.3 Study.5
5 Specific monitoring method.5
5.1 Requirements for number-based particle size distribution and mass concentration .5
5.2 Measurement of number-based particle size distribution .5
5.3 Mass concentration measurement .6
5.4 Inhalation exposure chamber .6
6 Assessment of results .7
7 Test report.7
Annex A (informative) Example of nanoparticle characterization for inhalation toxicity testing.9
Bibliography.17

ISO 10808:2010(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the
International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 10808 was prepared by Technical Committee ISO/TC 229, Nanotechnologies.

iv © ISO 2010 – All rights reserved

ISO 10808:2010(E)
Introduction
The number of nanotechnology-based consumer products containing silver, gold, carbon, zinc oxide, titanium
dioxide and silica nanoparticles is growing very rapidly. The population at risk of exposure to nanoparticles
continues to increase as the applications expand. In particular, workers in nanotechnology-based industries
are at risk of being exposed to nanoparticles. If nanoparticles are liberated from products, the public could be
exposed as well. Although toxicity screening using instillation of nanomaterials provides important information,
it does not reflect the actual scenario of inhalation exposure and does not provide the data required for
inhalation exposure risk assessment. In addition, while inhalation toxicology using rats is the norm at this time,
[10]
it is desirable to replace this antiquated method with a human-relevant assay .
The inhalation toxicity of nanoparticles is of particular concern in ensuring the health of workers and
consumers. In order to conduct inhalation toxicity studies of nano-sized particles, the monitoring of
concentration, size and distribution of nano-sized particles in the inhalation chamber is necessary. The
conventional methods of fine or coarse particle monitoring, such as weight-based mass dose monitoring, are
considered insufficient for nanoparticles, since nano-specific parameters (particle surface area, particle
number, etc.) might be critical determinants, and if so, should also be monitored.
This International Standard proposes a battery of inhalation toxicity testing chamber monitoring, including a
differential mobility analyzing system (DMAS), for measuring particle number, size, distribution, surface area
and estimated mass dose, as well as morphological examination using transmission electron microscopy
(TEM) or scanning electron microscopy (SEM) equipped with an energy dispersive X-ray analyzer
(TEM-EDXA) for chemical composition.
This International Standard also includes conventional mass dose monitoring and other physicochemical
monitoring, for use when deemed a necessary parameter for toxicity determination. This method evaluates
nano-sized particle surface area, mass dose, particle distribution, composition and dispersion to support
[13][17][18]
effective analysis of inhalation toxicity testing results .

INTERNATIONAL STANDARD ISO 10808:2010(E)

Nanotechnologies — Characterization of nanoparticles in
inhalation exposure chambers for inhalation toxicity testing
1 Scope
This International Standard specifies requirements for, and gives guidance on, the characterization of airborne
nanoparticles in inhalation exposure chambers for the purpose of inhalation toxicity studies in terms of particle
mass, size distribution, number concentration and composition.
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 10312, Ambient air —Determination of asbestos fibres — Direct transfer transmission electron
microscopy method
ISO 15900, Determination of particle size distribution — Differential electrical mobility analysis for aerosol
particles
ISO/TS 27687, Nanotechnologies — Terminology and definitions for nano-objects — Nanoparticle, nanofibre
and nanoplate
1)
OECD Test Guideline 403 (TG 403), Acute Inhalation Toxicity
1)
OECD Test Guideline 412 (TG 412), Subacute Inhalation Toxicity: 28-Day Study
1)
OECD Test Guideline 413 (TG 413), Subchronic Inhalation Toxicity: 90-Day Study
1)
OECD Guidance Document 39 (GD 39), Acute Inhalation Toxicity Testing
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 15900 and ISO/TS 27687 and the
following apply.
1) Organization for Economic Cooperation and Development (OECD) publication.
ISO 10808:2010(E)
3.1 Particle measuring systems
3.1.1
differential electrical mobility classifier
DEMC
differential electrical mobility spectrometer
DEMS
classifier that is able to select aerosol particle sizes from a distribution that enters it and pass only selected
sizes to the exit
NOTE 1 A DEMC classifies aerosol particle sizes by balancing the electrical force on each particle in an electrical field
with its aerodynamic drag force. Classified particles have different sizes due to their number of electrical charges and a
narrow range of electrical mobility determined by the operating conditions and physical dimensions of the DEMC.
NOTE 2 Adapted from ISO 15900:2009, definition 2.7.
3.1.2
differential mobility analyzing system
DMAS
system used to measure the size distribution of submicrometre aerosol particles consisting of a DEMC,
a particle charge conditioner, flow meters, a particle detector, interconnecting plumbing, a computer and
suitable software
NOTE Adapted from ISO 15900:2009, definition 2.8.
3.1.3
condensation particle counter
CPC
instrument that detects particles and that can be used to calculate particle number concentration given the
known flow rates into the detector
NOTE 1 The range of particles detected are usually smaller than several hundred nanometers and larger than a few
nanometers. A CPC is one possible detector for use with a DEMC.
NOTE 2 In some cases, a condensation particle counter may be called a condensation nucleus counter (CNC).
NOTE 3 Adapted from ISO 15900:2009, definition 2.5.
3.2
inhalation exposure chamber
inhalation chamber
exposure chamber
system prepared to expose experimental animals to an inhaled test substance of predetermined duration and
dose by either the nose-only or whole-body method
NOTE 1 The term “nose-only” is synonymous with “head-only” or “snout-only”.
NOTE 2 Adapted from OECD TG 403, 412, 413.
3.3
nanoparticle generation system
device used to make nanoparticle aerosol with controlled size distribution and concentration
3.4
breathing zone
location from which the experimental animal breathes
NOTE 1 For an unrestrained, non-caged animal, this will be the entire volume of the inhalation chamber. For a
restrained or caged animal, this will be the range of motion for the animal's nose. For a masked animal, this will be the
small volume in front of the nostrils.
2 © ISO 2010 – All rights reserved

ISO 10808:2010(E)
NOTE 2 The term “breathing zone” is used to ensure test atmosphere samples are obtained from the same location as
that in which the animal breathes. An undesirable sampling approach would be one where concentration measurements
are obtained at the top of the inhalation chamber while the animal is exposed at the bottom.
3.5
geometric mean diameter
GMD
measure of central tendency of particle size distribution using the logarithm of particle diameters, computed
for the DMAS by
n
ΔNdln
()
∑ ii
im=
ln(GMD)=
N
where
d is the midpoint diameter for the size channel, i;
i
N is the total concentration;
ΔN is the concentration within the size channel, i;
i
m is the first channel;
n is the last channel.
NOTE The GMD is normally computed from particle counts and when noted may be based on surface area or
particle volume with appropriate weighting.
3.6
geometric standard deviation
GSD
measure of width or spread of particle sizes, computed for the DMAS by
n 2
⎡⎤
Ndln − ln GMD
()
∑ ii
⎣⎦
im=
ln(GSD)=
N−1
3.7
count median diameter
CMD
diameter equal to GMD for particle counts assuming a logarithmic normal distribution
NOTE The general form of the relationship as described in ISO 9276-5 is
rp− s
()
CMD==xx e
50,rp50,
where
e is the base of natural logarithms, e = 2,718 28;
p is the dimensionality (type of quantity) of a distribution, where
p = 0 is the number,
p = 1 is the length,
p = 2 is the area, and
p = 3 is the volume or mass;
ISO 10808:2010(E)
r is the dimensionality (type of quantity) of a distribution, where
r = 0 is the number,
r = 1 is the length,
r = 2 is the area, and
r = 3 is the volume or mass;
s is the standard deviation of the density distribution;
x is the median particle size of a cumulative distribution of dimensionality, r.
50,r
4 Test substance monitoring method
4.1 Principle
4.1.1 Exposure
Precise characterization of the test substance exposure is essential for an inhalation toxicology study. The
objective in nanoparticle inhalation toxicology is to establish a quantitative relationship between the observed
toxicological outcome and the dose metrics used in terms of test substance physical and chemical properties.
4.1.2 Particle properties
The specific chemical and physical properties of the nanoparticle should be determined to the extent possible;
however, because these may not be known a priori, as many parameters as possible should be determined.
Nanoparticle composition, number and mass concentrations, median and mean size and size distribution,
surface area, electrical charge, surface character, hygroscopicity and shape might be important parameters
for dosimetry.
4.2 Preparation of system
4.2.1 During development of the nanoparticle generating system and prior to interfacing with the exposure
chamber(s), measurements should be performed to verify aerosol particle composition and purity and to
establish the stability. During exposure tests, analysis should be conducted continuously and/or intermittently
depending on the method of analysis to determine the consistency of particle size distribution without
disrupting the inhalation exposure.
[3]
NOTE A nanoparticle generating system for silver and other metals is described in ISO 10801 .
4.2.2 Inhalation chambers and supporting equipment shall be prepared in accordance with OECD TG 403,
OECD TG 412 and OECD TG 413.
4.2.3 Inhalation chambers and supporting equipment shall be prepared for nanoparticle exposure studies.
NOTE 1 Aerosolized nanoparticles can be deposited to walls by Brownian diffusion and particle size change due to
aggregation/agglomeration. This deposition process depends on the particle size, electrostatic charge, particle number
concentration and residence time. See standard texts on aerosol science, References [11], [19] and [20].
NOTE 2 Charge neutralization might be required, depending on the purpose of the study.
If charge distribution is considered a characterization requirement, this shall be specified and measured in the
study.
NOTE 3 To reduce deposition losses, conductive tubing of the minimum length practical to use with the tubing diameter
is selected to interface with instrumentation.
4 © ISO 2010 – All rights reserved

ISO 10808:2010(E)
4.2.4 An inhalation chamber or chambers and supporting equipment, such as sampling probes and
manifolds, shall be characterized to ensure compliance with OECD TG 403, OECD TG 412 and
OECD TG 413, for determining any sampling bias.
NOTE Sampling manifold consists of tubing, solenoid valves and/or other elements required for routing samples from
each chamber to online monitoring equipment.
4.2.5 Measurement instruments used in inhalation testing should be calibrated and/or tested in accordance
with ISO 15900.
The DMAS is usually calibrated at the factory and this should be documented in the report.
NOTE In addition, in the course of using the DMAS, it must be routinely calibrated as well.
4.3 Study
4.3.1 The study sh
...