SIST EN ISO 10993-10:2003

SLOVENSKI STANDARD
SIST EN ISO 10993-10:2003
01-marec-2003
1DGRPHãþD
SIST EN ISO 10993-10:2000
%LRORãNRRYUHGQRWHQMHPHGLFLQVNLKSULSRPRþNRYGHO3UHVNXVLGUDåHQMDLQ
]DNDVQMHQHSUHREþXWOMLYRVWL,62
Biological evaluation of medical devices - Part 10: Tests for irritation and delayed-type
hypersensitivity (ISO 10993-10:2002)
Biologische Beurteilung von Medizinprodukten - Teil 10: Prüfung auf Irritation und
Allergien vom verzögerten Typ (ISO 10993-10:2002)
Evaluation biologique des dispositifs médicaux - Partie 10: Essais d'irritation et
d'hypersensibilité retardée (ISO 10993-10:2002)
Ta slovenski standard je istoveten z: EN ISO 10993-10:2002
ICS:
11.100.20 %LRORãNRRYUHGQRWHQMH Biological evaluation of
PHGLFLQVNLKSULSRPRþNRY medical devices
SIST EN ISO 10993-10:2003 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------

EUROPEAN STANDARD
EN ISO 10993-10
NORME EUROPÉENNE
EUROPÄISCHE NORM
September 2002
ICS 11.100 Supersedes EN ISO 10993-10:1995
English version
Biological evaluation of medical devices - Part 10: Tests for
irritation and delayed-type hypersensitivity (ISO 10993-10:2002)
Evaluation biologique des dispositifs médicaux - Partie 10: Biologische Beurteilung von Medizinprodukten - Teil 10:
Essais d'irritation et d'hypersensibilité retardée (ISO 10993- Prüfung auf Irritation und Allergien vom verzögerten Typ
10:2002) (ISO 10993-10:2002)
This European Standard was approved by CEN on 5 August 2002.
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 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 Management Centre has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,
Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, 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
© 2002 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 10993-10:2002 E
worldwide for CEN national Members.

---------------------- Page: 2 ----------------------

EN ISO 10993-10:2002 (E)
CORRECTED 2002-11-27
Foreword
This document (EN ISO 10993-10:2002) has been prepared by Technical Committee ISO/TC
194 "Biological evaluation of medical devices" in collaboration with Technical Committee
CEN/TC 206 "Biocompatibility of medical and dental materials and devices", the secretariat of
which is held by NEN.
This document supersedes EN ISO 10993-10:1995.
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 March 2003, and conflicting national
standards shall be withdrawn at the latest by March 2003.
This document has been prepared under a mandate given to CEN by the European Commission
and the European Free Trade Association, and supports essential requirements of EU
Directive(s).
For relationship with EU Directive(s), see informative Annex ZB, which is an integral part of this
document.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of
the following countries are bound to implement this European Standard: Austria, Belgium, Czech
Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg,
Malta, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 10993-10:2002 has been approved by CEN as EN ISO 10993-10:2002 without
any modifications.
NOTE Normative references to International Standards are listed in Annex ZA (normative).
2

---------------------- Page: 3 ----------------------

EN ISO 10993-10:2002 (E)
Annex ZA
(normative)
Normative references to international publications
with their relevant European publications
This European Standard incorporates by dated or undated reference, provisions from other
publications. These normative references are cited at the appropriate places in the text and the
publications are listed hereafter. For dated references, subsequent amendments to or revisions of
any of these publications apply to this European Standard only when incorporated in it by
amendment or revision. For undated references the latest edition of the publication referred to
applies (including amendments).
NOTE Where an International Publication has been modified by common modifications, indicated by (mod.),
the relevant EN/HD applies.
Publication Year Title EN/HD Year
ISO 10993-1 1997 Biological evaluation of medical devices - EN ISO 10993-1 1997
Part 1: Evaluation and testing
ISO 10993-2 1992 Biological evaluation of medical devices - EN ISO 10993-2 1998
Part 2: Animal welfare requirements
ISO 10993-9 1999 Biological evaluation of medical devices - EN ISO 10993-9 1999
Part 9: Framework for identification and
quantification of potential degradation
products
ISO 10993-12 1996 Biological evaluation of medical devices - EN ISO 10993-12 1996
Part 12: Sample preparation and reference
materials
ISO 10993-13 1998 Biological evaluation of medical devices - EN ISO 10993-13 1998
Part 13: Identification and quantification of
degradation products from polymeric
medical devices
ISO 10993-14 2001 Biological evaluation of medical devices - EN ISO 10993-14 2001
Part 14: Identification and quantification of
degradation products from ceramics
ISO 10993-15 2000 Biological evaluation of medical devices - EN ISO 10993-15 2000
Part 15: Identification and quantification of
degradation products from metals and
alloys
3

---------------------- Page: 4 ----------------------

EN ISO 10993-10:2002 (E)
Annex ZB
(informative)
Clauses of this European Standard addressing essential requirements or
other provisions of EU Directives
This European Standard has been prepared under a mandate given to CEN by the European
Commission and the European Free Trade Association and supports essential requirements of
EU Directive 93/42/EEC.
WARNING Other requirements and other EU Directives may be applicable to the product(s)
falling within the scope of this standard.
The following clauses of this standard are likely to support requirements of Directive 93/42/EEC.
Compliance with these clauses of this standard provides one means of conforming with the
specific essential requirements of the Directive concerned and associated EFTA regulations.
Table ZB.1— Correspondence between this European Standard and EU Directive 93/42/EEC
Clause/subclause of this Corresponding Essential Comments
European Standard Requirement of Directive
93/42/EEC
6 7.1 of Annex I
7
B.2 7.1 of Annex I
B.3
B.4
B.5 7.2 of Annex I
B.6
B.7
4

---------------------- Page: 5 ----------------------

INTERNATIONAL ISO
STANDARD 10993-10
Second edition
2002-09-01


Biological evaluation of medical devices —
Part 10:
Tests for irritation and delayed-type
hypersensitivity
Évaluation biologique des dispositifs médicaux —
Partie 10: Essais d'irritation et d'hypersensibilité retardée




Reference number
ISO 10993-10:2002(E)
©
 ISO 2002

---------------------- Page: 6 ----------------------

ISO 10993-10:2002(E)
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 2002
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.ch
Web www.iso.ch
Printed in Switzerland

ii © ISO 2002 – All rights reserved

---------------------- Page: 7 ----------------------

ISO 10993-10:2002(E)
Contents Page
Foreword . iv
Introduction. vi
1 Scope. 1
2 Normative references. 1
3 Terms and definitions. 2
4 General principles — Step-wise approach . 3
5 Pretest considerations. 4
5.1 General . 4
5.2 Types of material. 4
5.3 Information on chemical composition . 4
5.4 Material characterization . 5
6 Irritation tests . 5
6.1 In vitro irritation tests . 5
6.2 Factors to be considered in design and selection of in vivo tests . 5
6.3 Animal skin irritation test . 6
6.4 Human skin irritation test. 10
7 Delayed hypersensitivity tests. 14
7.1 Choice of test. 14
7.2 Choice of test sample concentrations . 14
7.3 Other important factors affecting the outcome of the test . 14
7.4 Maximization test for delayed hypersensitivity . 15
7.5 Closed-patch test for delayed hypersensitivity . 18
8 Key factors in interpretation of test results. 20
Annex A (normative) Preparation of materials for irritation/sensitization testing. 21
Annex B (informative) Additional irritation tests.23
Annex C (informative) Background information. 41
Bibliography. 45

© ISO 2002 – All rights reserved iii

---------------------- Page: 8 ----------------------

ISO 10993-10:2002(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 3.
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 part of ISO 10993 may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 10993-10 was prepared by Technical Committee ISO/TC 194, Biological evaluation of medical devices.
This second edition cancels and replaces the first edition (ISO 10993-10:1995), which has been technically revised.
ISO 10993 consists of the following parts, under the general title Biological evaluation of medical devices:
 Part 1: Evaluation and testing
 Part 2: Animal welfare requirements
 Part 3: Tests for genotoxicity, carcinogenicity and reproductive toxicity
 Part 4: Selection of tests for interactions with blood
 Part 5: Tests for in vitro cytotoxicity
 Part 6: Tests for local effects after implantation
 Part 7: Ethylene oxide sterilization residuals
 Part 8: Selection and qualification of reference materials for biological tests
 Part 9: Framework for identification and quantification of potential degradation products
 Part 10: Tests for irritation and delayed-type hypersensitivity
 Part 11: Tests for systemic toxicity
 Part 12: Sample preparation and reference materials
 Part 13: Identification and quantification of degradation products from polymeric medical devices
 Part 14: Identification and quantification of degradation products from ceramics
 Part 15: Identification and quantification of degradation products from metals and alloys
iv © ISO 2002 – All rights reserved

---------------------- Page: 9 ----------------------

ISO 10993-10:2002(E)
 Part 16: Toxicokinetic study design for degradation products and leachables
 Part 17: Establishment of allowable limits for leachable substances
 Part 18: Chemical characterization of materials
Future parts will deal with other relevant aspects of biological testing.
This part of ISO 10993 is a harmonization of numerous standards and guidelines, including BS 5736, OECD
Guidelines, U.S. Pharmacopoeia and the European Pharmacopoeia. It is intended to be the basic document for the
selection and conduct of tests enabling evaluation of irritation and dermal sensitization responses relevant to safety
of medical materials and devices.
Annex A forms a normative part of this part of ISO 10993. Annexes B and C are for information only.

© ISO 2002 – All rights reserved v

---------------------- Page: 10 ----------------------

ISO 10993-10:2002(E)
Introduction
This part of ISO 10993 assesses possible contact hazards from chemicals released from medical devices that may
produce skin and mucosal irritation, eye irritation and delayed contact hypersensitivity
Some materials that are included in medical devices have been tested, and their skin or mucosal irritation or
sensitization potential has been documented. Other materials and their chemical components have not been tested
and may induce adverse effects when in contact with biological tissues. The manufacturer is thus obliged to
evaluate each device for potential adverse effects prior to marketing.
Traditionally, small animal tests are performed prior to testing on humans to help predict human response. More
recently, in vitro tests as well as human tests have been added as alternatives. Despite progress and considerable
effort in this direction, a review of findings suggests that currently no satisfactory in vitro test has been devised to
eliminate the requirement for in vivo testing. Where appropriate, the preliminary use of in vitro methods is
encouraged for screening purposes prior to animal testing. In order to reduce the number of animals used, this part
of ISO 10993 presents a step-wise approach, with review and analysis of test results at each stage. An animal test
is usually required prior to human testing.
It is intended that these studies be conducted using Good Laboratory Practice and comply with regulations related
to animal welfare. Statistical analysis of data is recommended and should be used whenever appropriate.
The tests included in this part of ISO 10993 are important tools for the development of safe products, provided that
these are executed and interpreted by trained personnel.

vi © ISO 2002 – All rights reserved

---------------------- Page: 11 ----------------------

INTERNATIONAL STANDARD ISO 10993-10:2002(E)

Biological evaluation of medical devices —
Part 10:
Tests for irritation and delayed-type hypersensitivity
1 Scope
This part of ISO 10993 describes the procedure for the assessment of medical devices and their constituent
materials with regard to their potential to produce irritation and delayed-type hypersensitivity.
This part of ISO 10993 includes
a) pretest considerations,
b) details of the test procedures, and
c) key factors for the interpretation of the results.
Instructions are given in annex A for the preparation of materials specifically in relation to the above tests.
Supplementary tests which are required specifically for devices used intradermally in the ocular, oral, rectal, penile
and vaginal areas are given in annex B.
2 Normative references
The following normative documents contain provisions which, through reference in this text, constitute provisions of
this part of ISO 10993. For dated references, subsequent amendments to, or revisions of, any of these publications
do not apply. However, parties to agreements based on this part of ISO 10993 are encouraged to investigate the
possibility of applying the most recent editions of the normative documents indicated below. For undated
references, the latest edition of the normative document referred to applies. Members of ISO and IEC maintain
registers of currently valid International Standards.
ISO 10993-1:1997, Biological evaluation of medical devices — Part 1: Evaluation and testing
ISO 10993-2, Biological evaluation of medical devices — Part 2: Animal welfare requirements
ISO 10993-9, Biological evaluation of medical devices — Part 9: Framework for identification and quantification of
potential degradation products
ISO 10993-12, Biological evaluation of medical devices — Part 12: Sample preparation and reference materials
ISO 10993-13, Biological evaluation of medical devices — Part 13: Identification and quantification of degradation
products from polymeric medical devices
ISO 10993-14 Biological evaluation of medical devices — Part 14: Identification and quantification of degradation
products from ceramics
ISO 10993-15, Biological evaluation of medical devices — Part 15: Identification and quantification of degradation
products from metals and alloys
ISO 10993-18, Biological evaluation of medical devices — Part 18: Chemical characterization of materials
© ISO 2002 – All rights reserved 1

---------------------- Page: 12 ----------------------

ISO 10993-10:2002(E)
ISO 14155-1, Clinical investigation of medical devices for human subjects — Part 1: General requirements
ISO 14155-2, Clinical investigation of medical devices for human subjects — Part 2: Clinical investigation plans
3 Terms and definitions
For the purposes of this part of ISO 10993, the terms and definitions given in ISO 10993-1 and the following apply.
3.1
allergen
sensitizer
substance/material which is capable of inducing specific hypersensitivity such that, on subsequent exposure to the
same substance/material characteristic, allergic effects are produced
3.2
blank liquid
solvent portion treated in the same manner as the identical solvent used for the preparation of test samples but
without test material, and which is intended for the determination of a background response of the solvent
3.3
challenge
elicitation
process following the induction phase in which the immunological effects of subsequent exposures in an individual
to the inducing material are examined
3.4
corrosion
slow destruction of the texture or material of a tissue
EXAMPLE The action of a strong irritant.
3.5
delayed-type hypersensitization
induction of specific T-cell mediated immunological memory for an allergen to which an individual is exposed,
resulting in a delayed-type hypersensitivity reaction after secondary contact with the allergen
3.6
dose
quantity to be administered to the test system at one time
3.7
erythema
reddening of the skin or mucous membrane
3.8
eschar
scab or discoloured slough of skin
3.9
induction
process that leads to the de novo generation of an altered state of immunological reactivity in an individual to a
specific material
3.10
irritant
agent that produces irritation
2 © ISO 2002 – All rights reserved

---------------------- Page: 13 ----------------------

ISO 10993-10:2002(E)
3.11
irritation
localized non-specific inflammatory response to single, repeated or continuous application of a substance/material
3.12
necrosis
death of one or more cells, or portion of tissue or organ, resulting in irreversible damage
3.13
negative control
material or substance which, when tested by the procedure described, demonstrates the suitability of the procedure
to yield a reproducible, appropriate negative, nonreactive or background response in the test system
3.14
oedema
swelling due to abnormal infiltration of fluid into the tissues
3.15
positive control
material or substance which, when tested by the procedure described, demonstrates the suitability of the procedure
to yield a reproducible, appropriate positive or reactive response in the test system
3.16
solvent
material or substance used to moisten, dilute, suspend, extract or dissolve the test substance material
EXAMPLES Chemical, vehicle, medium, etc.
3.17
test material
material, device, device portion or component thereof that is sampled for biological or chemical testing
3.18
test sample
extract or portion of the test material that is subjected to biological or chemical testing
3.19
ulceration
open sore representing loss of superficial tissue
4 General principles — Step-wise approach
The available methods for testing irritation and sensitization were developed specifically to detect skin irritation and
sensitization potential. Other types of adverse affect are generally not predicted by these tests.
This part of ISO 10993 requires a step-wise approach, which shall include one or more of the following:
a) characterization of test material, involving chemical characterization and analysis of the test sample according
to the general principles described in ISO 10993-9, ISO 10993-13, ISO 10993-14, ISO 10993-15 and
ISO 10993-18;
b) literature review, including an evaluation of chemical and physical properties, and information on the irritation
and sensitization potential of any product constituent as well as structurally related chemicals and materials;
c) consideration of in vitro tests in preference to in vivo tests, and replacement of the latter as new in vitro
methods become available and validated. At the present time there are no validated in vitro tests (other than
simple screens) to detect irritants or sensitizers.
d) in vivo animal tests;
© ISO 2002 – All rights reserved 3

---------------------- Page: 14 ----------------------

ISO 10993-10:2002(E)
NOTE Acute in vivo animal studies are undertaken to test for materials not already classified as severe irritants or
strong sensitizers by step a) or b). Materials that do not demonstrate an acute dermal irritation at single exposure may then
be further evaluated following repeated exposure.
A test of a positive-control substance for skin sensitization [7] shall be run at least every six months by the
testing laboratory to validate the test system and demonstrate a positive response.
e) non-invasive human tests/clinical trials.
If the material has been demonstrated not to be an irritant, a sensitizer or toxic in animals, studies on skin
irritation may then be considered in humans.
5 Pretest considerations
5.1 General
It is important to emphasise that pretest considerations may result in the conclusion that testing for irritation and/or
sensitization is not necessary.
The requirements given in clause 5 of 10993-1:1997 and in the subclauses below apply.
5.2 Types of material
5.2.1 Initial considerations
It shall be taken into consideration that, during manufacture and assembly of medical devices, additional chemical
components may be used as processing aids, e.g. lubricants or mould-release agents. In addition to the chemical
components of the starting material and manufacturing process aids, adhesive/solvent residues from assembly and
also sterilant residues or reaction products resulting from the sterilization process may be present in a finished
product. Whether these compounds pose a health hazard/risk depends on the leakage or degradation
characteristics of the finished products.
5.2.2 Ceramics, metals and alloys
These materials are normally less complex than polymers and biologically derived materials in terms of the number
of chemical constituents.
5.2.3 Polymers
These materials are normally chemically more complex than those in 5.2.1 in terms of composition. A number of
additives may be present and the completeness of polymerization may vary.
5.2.4 Biologically derived materials
These materials are inherently complex in their composition. They often also contain process residues, e.g.cross-
linkers and anti-microbial agents. Biological materials may not be consistent from sample to sample.
The methods in this part of ISO 10993 have not been designed for testing of biologically derived materials and may
therefore be less adequate. For example, the tests in this part of ISO 10993 do not consider cross-species
sensitization.
5.3 Information on chemical composition
5.3.1 General
Full qualitative data on the chemical constituents of the material shall be established. Where relevant to biological
safety, quantitative data shall also be obtained. If quantitative data are not obtained, the rationale shall be
documented and justified.
4 © ISO 2002 – All rights reserved

---------------------- Page: 15 ----------------------

ISO 10993-10:2002(E)
5.3.2 Existing data sources
Qualitative and quantitative information on the composition shall be obtained where possible from the supplier of
the starting material.
For polymers this often requires access to proprietary information; provision should be made for the transfer and
use of such confidential information.
Qualitative information about any additional processing additives (for example, mould-release agents) shall also be
obtained from appropriate members of the manufacturing chain, including converters and component
manufacturers.
In the absence of any data on composition, a literature study to establish the likely nature of the starting material
and any additives is recommended to assist in the selection of the most appropriate methods of analysis for the
material concerned.
NOTE The composition of ceramics, metals and alloys may be in accordance with ISO or American Society of Testing
Materials (ASTM) standards and/or may be specified by the user. Howe
...