prEN ISO 11138-6

SLOVENSKI STANDARD
01-julij-2026
Sterilizacija zdravstvenih izdelkov - Biološki indikatorji - 6. del: Biološki indikatorji
za sterilizacijske postopke z uparjenim vodikovim peroksidom (ISO/DIS 11138-
6:2026)
Sterilization of health care products - Biological indicators - Part 6: Biological indicators
for vaporized hydrogen peroxide sterilization processes (ISO/DIS 11138-6:2026)
Sterilisation von Produkten für die Gesundheitsfürsorge - Biologische Indikatoren - Teil 6:
Biologische Indikatoren für Sterilisationsverfahren mit Wasserstoffperoxid (ISO/DIS
11138-6:2026)
Stérilisation des produits de santé - Indicateurs biologiques - Partie 6: Indicateurs
biologiques pour la stérilisation au peroxyde d'hydrogène (ISO/DIS 11138-6:2026)
Ta slovenski standard je istoveten z: prEN ISO 11138-6
ICS:
11.080.01 Sterilizacija in dezinfekcija na Sterilization and disinfection
splošno in general
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

DRAFT
International
Standard
ISO/DIS 11138-6
ISO/TC 198
Sterilization of health care
Secretariat: ANSI
products — Biological indicators —
Voting begins on:
Part 6: 2026-05-11
Biological indicators for vaporized
Voting terminates on:
2026-08-03
hydrogen peroxide sterilization
processes
ICS: 11.080.01
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
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BEING ACCEPTABLE FOR INDUSTRIAL,
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PROVIDE SUPPORTING DOCUMENTATION.
Reference number
ISO/DIS 11138-6:2026(en)
DRAFT
ISO/DIS 11138-6:2026(en)
International
Standard
ISO/DIS 11138-6
ISO/TC 198
Sterilization of health care
Secretariat: ANSI
products — Biological indicators —
Voting begins on:
Part 6:
Biological indicators for vaporized
Voting terminates on:
hydrogen peroxide sterilization
processes
ICS: 11.080.01
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
© ISO 2026
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
STANDARDS MAY ON OCCASION HAVE TO
ISO/CEN PARALLEL PROCESSING
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BE CONSIDERED IN THE LIGHT OF THEIR
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NOTIFICATION OF ANY RELEVANT PATENT
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Website: www.iso.org
Published in Switzerland Reference number
ISO/DIS 11138-6:2026(en)
ii
ISO/DIS 11138-6:2026(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 General manufacturing requirements . 1
5 Specific manufacturing requirements . 1
6 Test organism, population and resistance . 2
6.1 Test organism .2
6.2 Population .2
6.3 Resistance .2
6.3.1 Determination of resistance .2
6.3.2 All Survive test .2
6.3.3 All Kill test . .2
6.3.4 Additional concentration levels .3
6.3.5 Testing for D-value.3
7 Culture conditions . 3
8 Carrier and packaging . 3
Annex A (normative) Method for determination of resistance to VH2O2 sterilization . 4
Annex B (informative) Summary of method development for the determination of biological
indicator resistance to vaporized hydrogen peroxide sterilization . 6
Annex C (informative) Quantity of H O solution to inject based on chamber volume .13
2 2
Annex D (normative) VH2O2 resistometer performance requirement specific to the VH2O2 BI .15
Bibliography . 17

iii
ISO/DIS 11138-6:2026(en)
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).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
patent(s) which may be required to implement this document. However, implementers are cautioned that
this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of 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 www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 198, Sterilization of health care products, in
collaboration with the European Committee for Standardization (CEN) Technical Committee CEN/TC 102,
Sterilizers and associated equipment for processing of medical devices, in accordance with the Agreement on
technical cooperation between ISO and CEN (Vienna Agreement).
A list of all parts in the ISO 11138 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
ISO/DIS 11138-6:2026(en)
Introduction
ISO 11138-1 specifies production, labelling, test methods and performance requirements for the
manufacture of biological indicators including inoculated carriers and suspensions intended for use in
validation and monitoring of sterilization processes. This document gives specific requirements for those
biological indicators intended for use in vaporized hydrogen peroxide (VH2O2) sterilization processes.
The ISO 11138 series represents the current “state-of-the-art” according to the experts representing
manufacturers, users and regulatory authorities involved in developing these documents. The intent is not
to promote the use of biological indicators where such use is not advised, but rather to provide common
requirements for the production of biological indicators that are known to be in use today.
Requirements for the validation and control of VH2O2 sterilization are provided in ISO 22441.
NOTE There can be other national or regional standards covering requirements for sterilization or biological
indicators.
Guidance on selection, use and interpretation of results when using biological indicators can be found in
ISO 11138-7.
v
DRAFT International Standard ISO/DIS 11138-6:2026(en)
Sterilization of health care products — Biological
indicators —
Part 6:
Biological indicators for vaporized hydrogen peroxide
sterilization processes
1 Scope
This document specifies requirements for test organisms, inoculated carriers, biological indicators and test
methods intended for use in assessing the performance of sterilizers and sterilization processes employing
vaporized hydrogen peroxide (VH2O2) as the sterilizing agent.
NOTE Requirements for validation and control of VH2O2 sterilization processes are provided in ISO 22441.
This document does not cover biological indicators used for hydrogen peroxide (H O ) biodecontamination
2 2
systems such as those used for rooms, enclosures or environmental spaces.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes
requirements 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 11138-1:2017, Sterilization of health care products — Biological indicators — Part 1: General requirements
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 11138-1 apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
4 General manufacturing requirements
The general manufacturing requirements of ISO 11138-1:2017, Clause 4 shall apply.
5 Specific manufacturing requirements
The specific manufacturing requirements of ISO 11138-1:2017, Clause 5 shall apply.

ISO/DIS 11138-6:2026(en)
6 Test organism, population and resistance
6.1 Test organism
6.1.1 The preferred test organisms shall be spores of Geobacillus stearothermophilus.
6.1.2 If a test organism other than Geobacillus stearothermophilus is used, the suitability of the resistance
of that test organism shall be determined.
NOTE Annex A of ISO 22441:2022 provides factors for consideration in selecting microorganisms.
6.2 Population
6.2.1 For inoculated carriers and biological indicators, the population shall be greater than or equal to 1,0
× 10 .
n
6.2.2 The population shall be stated with increments less than or equal to 0,1×10 per unit (e.g. per ml of
suspension, per inoculated carrier or per biological indicator).
6.2.3 The population shall be determined in accordance with ISO 11138-1:2017, Annex A.
6.3 Resistance
6.3.1 Determination of resistance
Resistance shall be determined per the procedure defined in Annex A.
NOTE See ISO 11138-7 for information on use and applications of BIs.
6.3.2 All Survive test
6.3.2.1 All Survive means a result from the culture of a group of inoculated carriers or biological indicators
where every test article is found to contain surviving microorganisms after exposure to the test cycle.
6.3.2.2 Meeting the All Survive test result criteria is a requirement of this document. All Survive test
results must be reported by the biological indicator manufacturer for each lot.
6.3.2.3 Not less than 50 replicates shall be used for the All Survive test. All replicates must be positive to
meet the requirement (except as noted in 6.3.2.4).
NOTE The number of units run per exposure depends on both the capacity and the operating characteristics of
the resistometer being used. It can be necessary to run several exposures at the survival time in order to test the total
number of units required.
6.3.2.4 For lots with a single negative biological indicator in a Survival Test, an additional 100 samples
(minimum) may be tested. If no additional unexpected results are obtained, the All Survive requirement is
confirmed.
6.3.3 All Kill test
6.3.3.1 All Kill means a result from the culture of a group of inoculated carriers or biological indicators
where the population of microorganisms from each test article is found to have been completely inactivated
after exposure to the test cycle.

ISO/DIS 11138-6:2026(en)
6.3.3.2 Completing the All Kill test is a requirement of this standard. Meeting the All Kill criteria (all units
negative) is not required. All Kill test results must be reported by the biological indicator manufacturer for
each lot.
6.3.3.3 Not less than 50 replicates shall be used for the All Kill test.
NOTE The number of units run per exposure depends on both the capacity and the operating characteristics of
the resistometer being used. It can be necessary to run several exposures at the kill time in order to test the total
number of units required.
6.3.3.4 If there are >10 positives, verification that the resistometer is working according to specification
shall be performed.
6.3.4 Additional concentration levels
Additional concentration levels that fall between the levels defined in Annex A and particularly A.3.13 and
A.3.14 can be tested and reported by the BI manufacturer.
6.3.5 Testing for D-value
In addition to the resistance determination requirements in 6.3.1, testing for D-value can be performed
according to the BI manufacturer’s recommended method. ISO 11138-1 provides further information on
methods.
7 Culture conditions
The culture condition requirements of ISO 11138-1:2017, Clause 7 apply.
8 Carrier and packaging
The suitability of the carrier and primary packaging materials for biological indicators for use in VH2O2
sterilization processes shall be demonstrated in accordance with the requirements of ISO 11138-1:2017, 5.2
and Annex B.
ISO/DIS 11138-6:2026(en)
Annex A
(normative)
Method for determination of resistance to VH2O2 sterilization
A.1 Principle
This method is intended to define reproducible test conditions for the evaluation of the resistance of
biological indicators (inoculated carriers) for assessing the performance of VH2O2 processes. This method
requires the use of a resistometer or other test sterilizer that is capable of precise measurement and process
control.
A.2 Materials
Hydrogen peroxide (H O ) solution: nominal 59 % weight/weight +/- 1 % by weight.
2 2
A.3 Test procedure
A.3.1 Chamber pre-conditioning: before initiating a test cycle, the sample holder and the sterilizer chamber
shall be equilibrated to the required operating temperature (50 °C ± 1 °C).
A.3.2 Pre-condition the biological indicators to 23 °C ± 2 °C.
A.3.3 Calculate the amount of nominal 59 % H O by weight solution to be injected into the exposure
2 2
chamber based on the volume of the sterilizer chamber.
NOTE See Annex C for information on the quantity of H O sterilant solution to be injected
2 2
A.3.4 Conditioning cycle: if applicable run one or more warm up cycles to bring the test system to steady
state including the sample holder before the first test cycle. No BI test samples or data collection required.
A.3.5 Load the BI test samples into the chamber in a manner which minimizes temperature loss.
A.3.6 Immediately after loading the BI test samples initiate the test cycle according to Table A.1.
Table A.1 — Test cycle conditions
Variable Value
Starting liquid H O concentration Target 59 % w/w ± 1 % w/w
2 2
Chamber operating temperature 50 °C ± 1 °C
Pre-exposure evacuation time + Hold Time <8 min
Evacuation pressure 0,07 kPa (±0,01 kPa)
0,5 Torr (±0,10 Torr)
Time to achieve set point pressure or concentration of ≤10 s
VH2O2 (come-up period)
VH2O2 exposure pressure – All Survive (see NOTE) Minimum 0,27 kPa ± 0,05 kPa
(Minimum 2 Torr ± 0,4 Torr)
VH2O2 exposure pressure – All Kill (See NOTE) Minimum 1,47 kPa ± 0,08 kPa
(Minimum 11 Torr ± 0,6 Torr)
ISO
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