SIST EN ISO 14161:2010

Sterilization of health care products - Biological indicators - Guidance for the selection, use and interpretation of results (ISO 14161:2009)

This International Standard provides guidance for the selection, use and interpretation of results from application of biological indicators when used in the development, validation and routine monitoring of sterilization processes. This International Standard applies to biological indicators for which International Standards exist.

Sterilisation von Produkten für die Gesundheitsfürsorge - Biologische Indikatoren - Leitfaden für die Auswahl, Verwendung und Interpretation von Ergebnissen (ISO 14161:2009)

Diese Internationale Norm enthält einen Leitfaden für die Auswahl, Verwendung und Interpretation
von Ergebnissen, die durch die Anwendung von biologischen Indikatoren bei der Entwicklung, Validierung und
Routineüberwachung von Sterilisationsverfahren erzielt werden. Diese Internationale Norm gilt für biologische
Indikatoren, für die Internationale Normen vorliegen.
ANMERKUNG 1 Siehe z. B. Normen der Reihe ISO 11138.
ANMERKUNG 2 Die allgemeinen Informationen dieser Internationalen Norm könnten auch nutzbringend auf Verfahren
und biologische Indikatoren anwendbar sein, für die zur Zeit keine Internationalen Normen zur Verfügung stehen, wie z. B.
neue und in Entwicklung befindliche Sterilisationsverfahren.
1.2 Diese Internationale Norm umfasst keine Verfahren, die ausschließlich auf physikalischer Entfernung
von Mikroorganismen, z. B. durch Filtration, beruhen.
1.3 Diese Internationale Norm gilt nicht für Kombinationsverfahren, die z. B. Reinigungs-/Desinfektionsgeräte
verwenden oder das Durchspülen und die Dampfbehandlung von Rohrleitungen.
1.4 Diese Internationale Norm gilt nicht für Sterilisationsverfahren mit Flüssigkeiten.

Stérilisation des produits de santé - Indicateurs biologiques - Directives générales pour la sélection, l'utilisation et l'interprétation des résultats (ISO 14161:2009)

L'ISO 14161:2009 fournit des directives pour la sélection, l'utilisation et l'interprétation des résultats issus de l'application des indicateurs biologiques dans le développement, la validation et le contrôle périodique des procédés de stérilisation.
L'ISO 14161:2009 s'applique aux indicateurs biologiques pour lesquels il existe des Normes internationales.

Sterilizacija izdelkov za zdravstveno nego - Biološki indikatorji - Navodilo za izbiro, uporabo in predstavitev rezultatov (ISO 14161:2009)

Ta mednarodni standard zagotavlja navodilo za izbiro, uporabo in predstavitev rezultatov delovanja bioloških indikatorjev, kadar se uporabljajo pri razvoju, validaciji in rutinskem spremljanju sterilizacijskih postopkov. Ta mednarodni standard velja za biološke indikatorje, za katere obstajajo mednarodni standardi.

General Information

• Status: Withdrawn
• Publication Date: 15-Dec-2009
• Withdrawal Date: 10-Jun-2019
• ICS: 11.080.01 - Sterilization and disinfection in general
• Technical Committee: VAZ - Healthcare
• Current Stage: 9900 - Withdrawal (Adopted Project)
• Start Date: 11-Jun-2019
• Due Date: 04-Jul-2019
• Completion Date: 11-Jun-2019
• Directive: 93/42/EEC - Council Directive 93/42/EEC of 14 June 1993 concerning medical devices
• Ref Project: EN ISO 14161:2009 - Sterilization of health care products - Biological indicators - Guidance for the selection, use and interpretation of results (ISO 14161:2009)

Relations

• Replaced By: SIST EN ISO 11138-7:2019 - Sterilization of health care products - Biological indicators - Part 7: Guidance for the selection, use and interpretation of results (ISO 11138-7:2019)
• Effective Date: 01-Jul-2019
• Replaces: SIST EN ISO 14161:2001 - Sterilization of health care products - Biological indicators - Guidance for the selection, use and interpretation of results (ISO 14161:2000)
• Effective Date: 01-Jan-2010

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 14161:2010
01-januar-2010
1DGRPHãþD
SIST EN ISO 14161:2001
Sterilizacija izdelkov za zdravstveno nego - Biološki indikatorji - Navodilo za izbiro,
uporabo in predstavitev rezultatov (ISO 14161:2009)
Sterilization of health care products - Biological indicators - Guidance for the selection,
use and interpretation of results (ISO 14161:2009)
Sterilisation von Produkten für die Gesundheitsfürsorge - Biologische Indikatoren -
Leitfaden für die Auswahl, Verwendung und Interpretation von Ergebnissen (ISO
14161:2009)
Stérilisation des produits de santé - Indicateurs biologiques - Directives générales pour la
sélection, l'utilisation et l'interprétation des résultats (ISO 14161:2009)
Ta slovenski standard je istoveten z: EN ISO 14161:2009
ICS:
11.080.01 Sterilizacija in dezinfekcija na Sterilization and disinfection
splošno in general
SIST EN ISO 14161:2010 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 14161:2010

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SIST EN ISO 14161:2010


EUROPEAN STANDARD
EN ISO 14161

NORME EUROPÉENNE

EUROPÄISCHE NORM
September 2009
ICS 11.080.01 Supersedes EN ISO 14161:2000
English Version
Sterilization of health care products - Biological indicators -
Guidance for the selection, use and interpretation of results (ISO
14161:2009)
Stérilisation des produits de santé - Indicateurs biologiques Sterilisation von Produkten für die Gesundheitsfürsorge -
- Directives générales pour la sélection, l'utilisation et Biologische Indikatoren - Leitfaden für die Auswahl,
l'interprétation des résultats (ISO 14161:2009) Verwendung und Interpretation von Ergebnissen (ISO
14161:2009)
This European Standard was approved by CEN on 31 July 2009.

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 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 Management Centre has the same status as the
official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, 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
© 2009 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 14161:2009: E
worldwide for CEN national Members.

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SIST EN ISO 14161:2010
EN ISO 14161:2009 (E)
Contents Page
Foreword .3

2

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SIST EN ISO 14161:2010
EN ISO 14161:2009 (E)
Foreword
This document (EN ISO 14161:2009) has been prepared by Technical Committee ISO/TC 198 "Sterilization of
health care products" in collaboration with Technical Committee CEN/TC 102 “Sterilizers for medical
purposes” the secretariat of which is held by DIN.
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 2010, and conflicting national standards shall be withdrawn at
the latest by March 2010.
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.
This document supersedes EN ISO 14161:2000.
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, 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 14161:2009 has been approved by CEN as a EN ISO 14161:2009 without any modification.

3

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SIST EN ISO 14161:2010

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SIST EN ISO 14161:2010

INTERNATIONAL ISO
STANDARD 14161
Second edition
2009-09-15

Sterilization of health care products —
Biological indicators — Guidance for the
selection, use and interpretation of
results
Stérilisation des produits de santé — Indicateurs biologiques —
Directives générales pour la sélection, l'utilisation et l'interprétation des
résultats




Reference number
ISO 14161:2009(E)
©
ISO 2009

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SIST EN ISO 14161:2010
ISO 14161:2009(E)
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ii © ISO 2009 – All rights reserved

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SIST EN ISO 14161:2010
ISO 14161:2009(E)
Contents Page
Foreword .v
Introduction.vi
1 Scope.1
2 Normative references.1
3 Terms and definitions .2
4 General .5
5 Characteristics of biological indicators .7
5.1 General .7
5.2 Test organism suspension for direct inoculation of products.7
5.3 Inoculated carriers .8
5.4 Self-contained biological indicators.8
5.5 Other biological indicators.9
6 Selection of supplier .9
6.1 General .9
6.2 Documentation .10
7 Biological indicators in process development.11
7.1 General .11
7.2 Overkill approach .12
7.3 Combined biological indicator and bioburden method.12
7.4 Bioburden method.13
8 Biological indicators in sterilization validation.14
8.1 General .14
8.2 Placement and handling of biological indicators .14
8.3 Sterilizer qualification .14
8.4 Performance qualification .14
8.5 Review and approval of validation .15
8.6 Requalification.15
9 Biological indicators in routine monitoring.15
9.1 General .15
9.2 Placement and handling of biological indicators .15
9.3 Process challenge device (PCD).16
10 Results.16
10.1 General .16
10.2 Interpretation of results .16
11 Application of biological indicator standards .17
11.1 General assessment of biological indicator performance by the user.17
11.2 Nominal population of test organism.17
11.3 Resistance determination.18
11.4 z value determination .20
11.5 F equivalent sterilization value determination .22
(T, z)
11.6 Establishing spore-log-reduction (SLR) .23
11.7 Sterility assurance level (SAL) calculation.23
11.8 Test equipment .24
12 Culture conditions.24
12.1 General .24
12.2 Incubation temperature.24
© ISO 2009 – All rights reserved iii

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SIST EN ISO 14161:2010
ISO 14161:2009(E)
12.3 Incubation period.25
12.4 Choice of growth medium.25
13 Third-party requirements .26
13.1 General.26
13.2 Minimum requirements for replicates and total number of biological indicators .26
13.3 Test equipment .26
14 Personnel training .27
15 Storage and handling .27
16 Disposal of biological indicators .27
Annex A (informative) Microbiological inactivation kinetics and enumeration techniques.28
Annex B (informative) Process challenge devices .34
Annex C (informative) Formulae for fraction negative methods for D value calculations .35
Annex D (informative) Examples of documentation for biological indicators prepared by the user .50
Annex E (informative) Calculation of z value.54
Annex F (informative) D value determination by survivor curve method.57
Annex G (informative) Survival-kill response characteristics .61
Bibliography .62

iv © ISO 2009 – All rights reserved

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SIST EN ISO 14161:2010
ISO 14161:2009(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 14161 was prepared by Technical Committee ISO/TC 198, Sterilization of health care products.
This second edition cancels and replaces the first edition (ISO 14161:2000), which has been technically
revised.
© ISO 2009 – All rights reserved v

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SIST EN ISO 14161:2010
ISO 14161:2009(E)
Introduction
This International Standard provides guidance regarding the selection, use and interpretation of results of
biological indicators when used to develop, validate and monitor sterilization processes. The procedures
described in this International Standard are of a general nature and do not, of themselves, constitute a
comprehensive development, validation or monitoring programme with regard to the sterilization of health care
products. The intent of this International Standard is not to mandate the use of biological indicators in a
process but, if they are used, to provide guidance for their proper selection and use in order to obviate
misleading results.
In this International Standard, users will find guidance on selection of the correct biological indicator for their
particular sterilization process and critical parameters as well as guidance on its appropriate use.
The user should select a biological indicator that is appropriate for the particular process to be used. There is
a wide variety of sterilization processes in common use, and biological indicator manufacturers are not able to
foresee all possible uses of their product. Manufacturers, therefore, label biological indicators according to
their intended use. It is the responsibility of the users of biological indicators to select, use, recover and
interpret the results as appropriate for the particular sterilization process used.
The certified performance of a biological indicator can be adversely affected by the conditions of storage and
transport prior to its use, by the use of the biological indicator or by the sterilizer process parameters. In
addition, the incubation procedure used after exposure to the process, including outgrowth temperature and
culture medium type, supplier and specific lot, can affect measured resistance as a function of recovery and
growth. For these reasons, the recommendations of the biological indicator manufacturer for storage and use
should be followed. After exposure, biological indicators should be aseptically transferred (if applicable) and
incubated as specified by the biological indicator manufacturer.
It should be noted that biological indicators are not intended to indicate that the products in the load being
sterilized are sterile. Biological indicators are utilized to test the effectiveness of a given sterilization process
and the equipment used, by assessing microbial lethality according to the concept of sterility assurance level.
Suitably trained personnel should conduct these studies.

vi © ISO 2009 – All rights reserved

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SIST EN ISO 14161:2010
INTERNATIONAL STANDARD ISO 14161:2009(E)

Sterilization of health care products — Biological indicators —
Guidance for the selection, use and interpretation of results
1 Scope
This International Standard provides guidance for the selection, use and interpretation of results from
application of biological indicators when used in the development, validation and routine monitoring of
sterilization processes. This International Standard applies to biological indicators for which International
Standards exist.
NOTE 1 See, for example, the ISO 11138 series.
NOTE 2 The general information provided in this International Standard can have useful application for processes and
biological indicators not currently addressed by existing International Standards, e.g., new and developing sterilization
processes.
This International Standard does not consider those processes that rely solely on physical removal of
microorganisms, e.g., filtration.
This International Standard is not intended to apply to combination processes using, for example, washer
disinfectors or flushing and steaming of pipelines.
This International Standard is not intended to apply to liquid sterilization processes.
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 11135-1, Sterilization of health care products — Ethylene oxide — Part 1: Requirements for development,
validation and routine control of a sterilization process for medical devices
ISO 11138-1:2006, Sterilization of health care products — Biological indicators — Part 1: General
requirements
ISO 11138-2, Sterilization of health care products — Biological indicators — Part 2: Biological indicators for
ethylene oxide sterilization processes
ISO 11138-3, Sterilization of health care products — Biological indicators — Part 3: Biological indicators for
moist heat sterilization processes
ISO 11138-4, Sterilization of health care products — Biological indicators — Part 4: Biological indicators for
dry heat sterilization processes
ISO 11138-5, Sterilization of health care products — Biological indicators — Part 5: Biological indicators for
low-temperature steam and formaldehyde sterilization processes
ISO 11737-1, Sterilization of medical devices — Microbiological methods — Part 1: Determination of a
population of microorganisms on products
© ISO 2009 – All rights reserved 1

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SIST EN ISO 14161:2010
ISO 14161:2009(E)
ISO 14937, Sterilization of health care products — General requirements for characterization of a sterilizing
agent and the development, validation and routine control of a sterilization process for medical devices
ISO 17665-1, Sterilization of health care products — Moist heat — Part 1: Requirements for the development,
validation and routine control of a sterilization process for medical devices
ISO 18472:2006, Sterilization of health care products — Biological and chemical indicators — Test equipment
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
accreditation
procedure by which an authoritative body gives formal recognition that a body or person is competent to carry
out specific tasks
[3]
NOTE 1 See ISO/IEC 17011 .
NOTE 2 Accreditation does not itself qualify the laboratory to approve any particular product. However, accreditation
can be relevant to approval and certification authorities when they decide whether or not to accept data produced by a
given laboratory in connection with their own activities.
3.2
aseptic technique
conditions and procedures used to exclude the introduction of microbial contamination
3.3
bioburden
population of viable microorganisms on or in a product and/or sterile barrier system
[ISO/TS 11139, definition 2.2]
3.4
biological indicator
BI
test system containing viable microorganisms providing a defined resistance to a specified sterilization
process
[ISO/TS 11139, definition 2.3]
3.5
D value
D value
10
time or dose required to achieve inactivation of 90 % of a population of the test microorganism under stated
conditions
[ISO/TS 11139, definition 2.11]
3.6
holding time
period for which the sterilization variable within the sterilizer and at all points within the load are continuously
within the limits specified for the sterilization stage
3.7
inoculated carrier
supporting material on or in which a defined number of viable test organisms have been deposited
NOTE 1 See ISO 11138-1.
NOTE 2 The test organism is a microorganism used for the manufacture of inoculated carriers.
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SIST EN ISO 14161:2010
ISO 14161:2009(E)
3.8
installation qualification
IQ
process of obtaining and documenting evidence that equipment has been provided and installed in
accordance with its specification
[ISO/TS 11139, definition 2.22]
3.9
inoculation
addition of a defined amount of a characterized microbial entity into or on to an item
3.10
log reduction
LR
reduction in number of viable microorganisms, expressed in log units
3.11
operational qualification
OQ
process of obtaining and documenting evidence that installed equipment operates within predetermined limits
when used in accordance with its operational procedures
[ISO/TS 11139, definition 2.27]
3.12
performance qualification
PQ
process of obtaining and documenting evidence that the equipment, as installed and operated in accordance
with operational procedures, consistently performs in accordance with predetermined criteria and thereby
yields product meeting its specification
[ISO/TS 11139, definition 2.30]
3.13
process challenge device
PCD
item designed to constitute a defined resistance to a sterilization process and used to assess performance of
the process
[ISO/TS 11139, definition 2.33]
3.14
process challenge location
PCL
site that simulates “worst case” conditions as they are given for sterilizing agent(s) in the goods to be sterilized
3.15
process parameter
specified value for a process variable
NOTE The specification for a sterilization process includes the process parameters and their tolerances.
[ISO/TS 11139, definition 2.34]
3.16
process variable
condition within a sterilization process, changes in which alter microbicidal effectiveness
EXAMPLE Time, temperature, pressure, concentration, humidity, wavelength.
[ISO/TS 11139, definition 2.35]
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SIST EN ISO 14161:2010
ISO 14161:2009(E)
3.17
reference microorganism
microbial strain obtained from a recognized culture collection
[ISO/TS 11139, definition 2.39]
3.18
resistometer
test equipment designed to create defined reference combinations of the physical and/or chemical variables of
a sterilization process
NOTE 1 Adapted from ISO 11138-1, definition 3.15 and ISO 18472:2006, definition 3.11.
NOTE 2 Also referred to as Biological Indicator Evaluator Resistometer (BIER).
3.19
spore-log-reduction
SLR
log of initial spore population, N , minus the log of the final population, N
0 F
3.20
sterile
free from viable microorganisms
[ISO/TS 11139, definition 2.43]
3.21
sterility assurance level
SAL
probability of a single viable microorganism occurring on an item after sterilization
−6 −3
NOTE The term SAL takes a quantitative value, generally 10 or 10 . When applying this quantitative value to
−6 −3
assurance of sterility, an SAL of 10 has a lower value but provides a greater assurance of sterility than an SAL of 10 .
[ISO/TS 11139, definition 2.46]
3.22
sterilization
validated process used to render product free from viable microorganisms
NOTE In a sterilization process, the nature of microbial inactivation is exponential and thus the survival of a
microorganism on an individual item can be expressed in terms of probability. While this probability can be reduced to a
very low number, it can never be reduced to zero.
[ISO/TS 11139, definition 2.47]
3.23
sterilization cycle development
procedure for determination of the appropriate processing parameters that are consistent with attaining the
desired specifications and label claims for a given product or group of products
3.24
sterilization cycle validation
documented procedure for obtaining, recording and interpreting the results required to establish that a process
will consistently yield product complying with predetermined specifications
3.25
supplier
organization or person that provides a product
EXAMPLE Producer, distributer, retailer or vendor of a product, or provider of a service or information.
4 © I
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