Chemical disinfectants and antiseptics - Quantitative test for the evaluation of bactericidal and yeasticidal and/or fungicidal activity of chemical disinfectants in the medical area on non-porous surfaces without mechanical action - Test method and requirements (phase 2, step 2)

This European Standard specifies a test method and the minimum requirements for bactericidal and/or yeasticidal  and/or fungicidal activity of chemical disinfectant products that form a homogeneous, physically stable preparation when diluted with hard water – or in the case of ready-to-use products – with water.
This European Standard applies to products that are used in the medical area for disinfecting non-porous surfaces without mechanical action.
This European Standard applies to areas and situations where disinfection is medically indicated. Such indications occur in patient care, for example:
in hospitals, in community medical facilities and in dental institutions;
in clinics of schools, of kindergartens and of nursing homes;
and may occur in the workplace and in the home. It may also include services such as laundries and kitchens supplying products directly for the patients.
NOTE   This method corresponds to a phase 2, step 2. test.
EN 14885 specifies in detail the relationship of the various tests to one another and to “use recommendations.

Chemische Desinfektionsmittel und Antiseptika - Quantitativer Versuch zur Bestimmung der bakteriziden und levuroziden und/oder fungiziden Wirkung chemischer Desinfektionsmittel im humanmedizinischen Bereich auf nicht porösen Oberflächen ohne mechanische Einwirkung - Prüfverfahren und Anforderungen (Phase 2, Stufe 2)

Dieses Dokument legt ein Prüfverfahren und die Mindestanforderungen an die bakterizide und levurozide und zusätzlich auch fungizide Wirkung von chemischen Desinfektionsmitteln fest, die bei Verdünnung mit Wasser standardisierter Härte als homogene, physikalisch stabile Zubereitung vorliegen bzw. bei gebrauchs¬fertigen Produkten mit Wasser verdünnt werden.
ANMERKUNG   Verdünnungen sind erforderlich, da drei Konzentrationen im aktiven bis nichtaktiven Bereich geprüft werden.
Dieses Dokument gilt für Produkte, die ohne mechanische Einwirkung zur Desinfektion nichtporöser Oberflächen im humanmedizinischen Bereich verwendet werden.
Dieses Dokument gilt für Bereiche und unter Bedingungen, wo eine Desinfektion oder Antiseptik aus medizinischen Gründen angezeigt ist. Indikationen dieser Art liegen z. B. bei der Patientenbetreuung in:
-   Krankenhäusern, kommunalen medizinischen Einrichtungen und im Dentalbereich;
-   medizinischen Einrichtungen in Schulen, Kindergärten und Heimen;
vor und können auch am Arbeitsplatz und im häuslichen Bereich gegeben sein. Eingeschlossen sein können auch Einrichtungen wie Wäschereien und Küchen, die der direkten Versorgung der Patienten dienen.
EN 14885 legt im Einzelnen die Beziehungen der verschiedenen Prüfungen untereinander und zu den Anwendungsempfehlungen fest.
Mit diesem Dokument ist es möglich, die Wirkung von Produkten wie handelsüblichen Formulierungen oder Wirkstoffen unter den Bedingungen, unter denen sie verwendet werden, auf Bakterien und/oder Pilze zu bestimmen und entspricht somit einer Prüfung der Phase 2, Schritt 2.
Dieses Verfahren schließt die Bewertung der Wirkung von Produkten gegen Mykobakterien und bakterielle Sporen aus.

Antiseptiques et désinfectants chimiques - Essai quantitatif pour l’évaluation de l’activité bactéricide et levuricide et/ou fongicide des désinfectants chimiques utilisés en médecine sur des surfaces non poreuses sans action mécanique - Méthode d’essai et exigences (phase 2, étape 2)

Le présent document spécifie une méthode d’essai et les exigences minimales relatives à l’activité bactéricide, levuricide ainsi que fongicide des désinfectants chimiques qui forment une préparation homogène, physiquement stable, lorsqu’ils sont dilués dans l’eau dure ou - dans le cas de produits prêts à l’emploi - dans l’eau.
NOTE   Des dilutions sont nécessaires car trois concentrations dont une concentration active et une concentration non active sont soumises à essai.
Le présent document s’applique aux produits utilisés en médecine pour désinfecter les surfaces non poreuses sans action mécanique.
Le présent document s’applique aux zones et aux situations où la désinfection ou l’antisepsie est médicalement préconisée. De telles indications se rencontrent dans le cadre des soins apportés aux patients, par exemple :
-   dans des hôpitaux, centres de soins médicaux et cabinets dentaires ;
-   dans des infirmeries d’écoles, de jardins d’enfants et de maisons de retraite ;
et peuvent aussi se rencontrer sur les lieux de travail ou à domicile. Elles peuvent également concerner des services tels que des blanchisseries et des cuisines qui fournissent des produits directement aux patients.
L’EN 14885 précise de manière détaillée la relation entre les différents essais et les recommandations d’utilisation.
Il est possible, en utilisant le présent document, de déterminer l’activité de produits tels que les formulations ou substances actives commerciales sur les bactéries et/ou champignons dans leurs conditions d’utilisation, ce qui correspond à un essai de phase 2, étape 2.
Cette méthode exclut l’évaluation de l’activité de produits sur les mycobactéries et les spores de bactéries.

Kemična razkužila in antiseptiki - Kvantitativni preskus na neporoznih površinah brez mehanskega delovanja za vrednotenje baktericidnega in/ali fungicidnega delovanja in delovanja kemičnih razkužil na kvasovke v humani medicini - Preskusna metoda in zahteve (faza 2, stopnja 2)

General Information

Status
Published
Public Enquiry End Date
01-Aug-2019
Publication Date
12-Dec-2021
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
04-Mar-2021
Due Date
09-May-2021
Completion Date
13-Dec-2021

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SLOVENSKI STANDARD
SIST EN 17387:2022
01-januar-2022
Kemična razkužila in antiseptiki - Kvantitativni preskus na neporoznih površinah
brez mehanskega delovanja za vrednotenje baktericidnega in/ali fungicidnega
delovanja in delovanja kemičnih razkužil na kvasovke v humani medicini -
Preskusna metoda in zahteve (faza 2, stopnja 2)
Chemical disinfectants and antiseptics - Quantitative test for the evaluation of

bactericidal and yeasticidal and/or fungicidal activity of chemical disinfectants in the

medical area on non-porous surfaces without mechanical action - Test method and
requirements (phase 2, step 2)

Chemische Desinfektionsmittel und Antiseptika - Quantitativer Versuch zur Bestimmung

der bakteriziden und levuroziden und/oder fungiziden Wirkung chemischer

Desinfektionsmittel im humanmedizinischen Bereich auf nicht porösen Oberflächen ohne

mechanische Einwirkung - Prüfverfahren und Anforderungen (Phase 2, Stufe 2)

Antiseptiques et désinfectants chimiques - Essai quantitatif pour l’évaluation de l’activité

bactéricide et levuricide et/ou fongicide des désinfectants chimiques utilisés en

médecine sur des surfaces non poreuses sans action mécanique - Méthode d’essai et

exigences (phase 2, étape 2)
Ta slovenski standard je istoveten z: EN 17387:2021
ICS:
11.080.20 Dezinfektanti in antiseptiki Disinfectants and antiseptics
SIST EN 17387:2022 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN 17387:2022
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SIST EN 17387:2022
EN 17387
EUROPEAN STANDARD
NORME EUROPÉENNE
August 2021
EUROPÄISCHE NORM
ICS 11.080.20; 71.100.35
English Version
Chemical disinfectants and antiseptics - Quantitative test
for the evaluation of bactericidal and yeasticidal and/or
fungicidal activity of chemical disinfectants in the medical
area on non-porous surfaces without mechanical action -
Test method and requirements (phase 2, step 2)

Antiseptiques et désinfectants chimiques - Essai Chemische Desinfektionsmittel und Antiseptika -

quantitatif pour l'évaluation de l'activité bactéricide et Quantitativer Versuch zur Bestimmung der

levuricide et/ou fongicide des désinfectants chimiques bakteriziden und levuroziden und/oder fungiziden

utilisés en médecine sur des surfaces non poreuses Wirkung chemischer Desinfektionsmittel im

sans action mécanique - Méthode d'essai et exigences humanmedizinischen Bereich auf nicht porösen

(phase 2, étape 2) Oberflächen ohne mechanische Einwirkung -
Prüfverfahren und Anforderungen (Phase 2, Stufe 2)
This European Standard was approved by CEN on 25 January 2021.

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, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and

United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels

© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17387:2021 E

worldwide for CEN national Members.
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SIST EN 17387:2022
EN 17387:2021 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms, definitions and abbreviations ..................................................................................................... 5

3.1 Terms and definitions ................................................................................................................................... 5

3.2 Symbols and abbreviations ......................................................................................................................... 6

4 Requirements ................................................................................................................................................... 6

5 Test methods .................................................................................................................................................... 7

5.1 Principle ............................................................................................................................................................. 7

5.2 Materials and reagents .................................................................................................................................. 8

5.2.1 Test organisms ................................................................................................................................................. 8

5.2.2 Culture media and reagents ........................................................................................................................ 8

5.2.3 Test surface .................................................................................................................................................... 11

5.3 Apparatus and glassware .......................................................................................................................... 12

5.3.1 General ............................................................................................................................................................. 12

5.3.2 Usual microbiological laboratory equipment and, in particular, the following: .................. 12

5.4 Preparation of test organism suspensions and product test solutions .................................... 13

5.4.1 Test organism suspensions ...................................................................................................................... 13

5.4.2 Product test solutions ................................................................................................................................. 19

5.5 Procedure for assessing the bactericidal, yeasticidal and fungicidal activity of the

product ............................................................................................................................................................. 19

5.5.1 Choice of experimental conditions ........................................................................................................ 19

5.5.2 Test procedure .............................................................................................................................................. 20

5.5.3 Counting of the test mixtures ................................................................................................................... 23

5.6 Calculation and expression of results ................................................................................................... 24

5.6.1 Calculation of data ....................................................................................................................................... 24

5.6.2 Verification of methodology ..................................................................................................................... 25

5.6.3 Expression of results ................................................................................................................................... 25

5.6.4 Conclusion....................................................................................................................................................... 26

5.7 Test report ...................................................................................................................................................... 26

Annex A (informative) Corresponding referenced strains ........................................................................ 28

Annex B (informative) Neutralizers ................................................................................................................... 29

Annex C (informative) Expression of results with the dilution-neutralization method ................. 31

Annex D (informative) Bactericidal activity on surfaces under clean conditions ............................. 33

Annex E (informative) Vacuum drying by desiccator - Instruction of use ............................................ 35

Annex F (informative) Precision of the test result ........................................................................................ 37

Bibliography ................................................................................................................................................................. 38

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SIST EN 17387:2022
EN 17387:2021 (E)
European foreword

This document (EN 17387:2021) has been prepared by Technical Committee CEN/TC 216 “Chemical

disinfectants and antiseptics”, the secretariat of which is held by AFNOR.

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 February 2022, and conflicting national standards

shall be withdrawn at the latest by February 2022.

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. CEN shall not be held responsible for identifying any or all such patent rights.

Any feedback and questions on this document should be directed to the users’ national standards body.

A complete listing of these bodies can be found on the CEN website.

According to the CEN-CENELEC Internal Regulations, the national standards organisations 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, Republic of

North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the

United Kingdom.
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SIST EN 17387:2022
EN 17387:2021 (E)
Introduction

This document describes a surface test method for establishing whether a chemical disinfectant in the

area and fields described in the scope (Clause 1) has or does not have bactericidal and/or fungicidal or

yeasticidal activity on non-porous surfaces without mechanical action.

The laboratory test closely simulates practical conditions of application. Chosen conditions (contact

time, temperature, and microorganisms on surfaces...) reflect parameters which are found in practical

situations including conditions which can influence the action of disinfectants.

However, for some applications the recommendations of use of a product can differ and therefore

additional test conditions need to be used.

This document is not intended to be used when product is applied via an automatic airborne

disinfection method; in such cases, see EN 17272.
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SIST EN 17387:2022
EN 17387:2021 (E)
1 Scope

This document specifies a test method and the minimum requirements for bactericidal and yeasticidal

and additionally fungicidal activity of chemical disinfectant products that form a homogeneous,

physically stable preparation when diluted with hard water – or in the case of ready-to-use products –

with water.

NOTE Dilutions are necessary as three concentrations in the active to non-active range are tested.

This document applies to products that are used in the medical area for disinfecting non-porous

surfaces without mechanical action.

This document applies to areas and situations where disinfection or antisepsis is medically indicated.

Such indications occur in patient care, for example:
— in hospitals, in community medical facilities and in dental institutions;

— in clinics of schools, of kindergartens and of nursing homes; and can occur in the workplace and in

the home. It can also include services such as laundries and kitchens supplying products directly for

the patients.

EN 14885 specifies in detail the relationship of the various tests to one another and to use

recommendations.

Using this document, it is possible to determine the activity of products like commercial formulations or

active substances on bacteria and/or fungi in the conditions in which they are used and therefore it

corresponds to a phase 2, step 2 test.

This method excludes the evaluation of the activity of products against mycobacteria and bacterial

spores.
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.

EN 12353, Chemical disinfectants and antiseptics - Preservation of test organisms used for the

determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal

(including bacteriophages) activity

EN 14885, Chemical disinfectants and antiseptics - Application of European Standards for chemical

disinfectants and antiseptics
3 Terms, definitions and abbreviations
3.1 Terms and definitions

For the purposes of this document, the terms and definitions given in EN 14885 apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at https://www.iso.org/obp
— IEC Electropedia: available at http://www.electropedia.org/
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SIST EN 17387:2022
EN 17387:2021 (E)
3.2 Symbols and abbreviations
c is the sum of V -values taken into account
cfu colony forming units
d is the dilution taken into account, lower dilution factor
m, m’ are the two replicas at the lower dilution expressed as cfu;
n, n’ are the two replicas at the higher dilution expressed as cfu;
n is the number of V -values taken into account
N number of cells per ml in the test suspension
N lg number of cfu recovered from the test surface in the water control
N counting of the cfu in the neutralizer control
N lg number of cfu recovered from the test surface in the test
N counting of the cfu in the method validation
Nts number of colony forming units remaining on the test surface
R reduction
V is the volume of the inoculated into the plate expressed in ml
4 Requirements

The product shall demonstrate at least a 5 decimal log (lg) reduction for bacteria and at least a 4

decimal log (lg) reduction for fungi, when tested in accordance with Table 1 and 5.5.

NOTE The 5 lg reduction is deemed necessary for the medical area to minimize the risk for the patients.

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SIST EN 17387:2022
EN 17387:2021 (E)
Table 1 — Experimental conditions
Test conditions Bactericidal activity Yeasticidal activity Fungicidal activity
Staphylococcus aureus
Minimum
Aspergillus brasiliensis
spectrum of Enterococcus hirae Candida albicans
Candida albicans
test organisms
Pseudomonas aeruginosa
additional any relevant test organism
according to the manufacturer’s recommendation, but between
Test
(4 ± 1) °C to (30 ± 1) °C
temperature
For tests performed at room temperature, the range shall be (21,5 ± 3,5) °C
according to the manufacturer’s recommendation, but at minimum 1 min
Contact time

and no longer than 5 min or 60 min (from 1 min to 5 min at intervals of 1 min and

from 5 min to 60 min at intervals of 5 min)
Interfering substance
clean conditions 0,3 g/l bovine albumin
dirty conditions 3,0 g/l bovine albumin plus 3,0 ml/l erythrocytes
additional any relevant substance

The contact times for surface disinfectants stated in this table are chosen on the basis of the practical

conditions of the product. The recommended contact time for the use of the product is within the responsibility

of the manufacturer. Products intended to disinfect surfaces that are likely to come into contact with the patient

and / or the medical staff and surfaces, which are frequently touched by different people, leading to the

transmission of microorganisms to the patient, shall be tested with a contact time of maximum 5 min. The same

applies where the contact time of the product shall be limited for practical reasons. Products for other surfaces

than stated above may be tested with a contact time of maximum 60 min.

NOTE For the additional conditions, the concentration defined as a result can be lower than the one obtained

under the minimum test conditions.
5 Test methods
5.1 Principle

A test suspension of bacteria or fungi in a solution of interfering substances (5.2.2.8) is inoculated onto

a test stainless steel surface and dried. A sample of the product under test is applied in a manner that

completely covers the dried test organisms. The surface is maintained at the temperature and contact

time exemplified in Clause 4 and 5.5.1. At the end of the contact time the surface is transferred to a

previously validated neutralization medium to suppress any product activity. The number of surviving

test organisms which can be recovered from the surface is determined quantitatively.

The number of bacteria or fungi on a test surface treated with hard water (or water in case of ready to

use products) in place of the product under test is also determined and the reduction in viable counts

attributed to the product is calculated as the difference between the two test surfaces’ results.

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SIST EN 17387:2022
EN 17387:2021 (E)
5.2 Materials and reagents
5.2.1 Test organisms
The bactericidal activity shall be evaluated using the following strains :
— Staphylococcus aureus ATCC 6 538;
— Enterococcus hirae ATCC 10 541;
— Pseudomonas aeruginosa ATCC 15 442.
The yeasticidal activity shall be evaluated using the following strain :
— Candida albicans ATCC 10 231.
The fungicidal activity shall be evaluated using the following strains :
— Aspergillus brasiliensis ATCC 16 404;
— Candida albicans ATCC 10 231.
If required for specific applications, additional strains may be chosen.

NOTE See Annex A for corresponding strain numbers in some other culture collections.

If additional test organisms are used, they shall be incubated under optimum growth conditions

(temperature, time, atmosphere, media) noted in the test report. If the additional test organisms

selected do not correspond to the specified strains, their suitability for supplying the required inocula

shall be verified. If these additional test organisms are not classified at a reference centre, their

identification characteristics shall be stated. In addition, they shall be held by the testing laboratory or

national culture collection under a reference for five years.
5.2.2 Culture media and reagents
5.2.2.1 General

All weights of chemical substances given in this document refer to the anhydrous salts. Hydrated forms

may be used as an alternative, but the weights required shall be adjusted to allow for consequent

molecular weight differences.

The reagents shall be of analytical grade and/or appropriate for microbiological purposes. They shall be

free from substances that are toxic or inhibitory to the test organisms.

To improve reproducibility, it is recommended that commercially available dehydrated material is used

for the preparation of culture media. The manufacturer's instructions relating to the preparation of

these products should be rigorously followed.
For each culture medium and reagent, a time limitation for use should be fixed.
All specified pH values are measured at (20 ± 1) °C.

The ATCC numbers are the collection numbers of strains supplied by these culture collections. This

information is given for the convenience of users of this European Standard and does not constitute an

endorsement by CEN of the product named.
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SIST EN 17387:2022
EN 17387:2021 (E)
5.2.2.2 Water

The water shall be freshly glass-distilled water and not demineralized water. If distilled water of

adequate quality is not available, water for injections can be used.

Sterilize in the autoclave [5.3.2.1a)]. Sterilization is not necessary if the water is used e.g. for

preparation of culture media and subsequently sterilized.
NOTE See 5.2.2.7 for the procedure to prepare hard water.
5.2.2.3 Tryptone soya agar (TSA)
For maintenance of bacterial strains and performance of viable counts.
Tryptone, pancreatic digest of casein 15,0 g
Soya peptone, papaic digest of Soybean meal 5,0 g
Sodium chloride (NaCl) 5,0 g
Agar 15,0 g
Water (5.2.2.2) to 1 000,0 ml

Sterilize in the autoclave [5.3.2.1 a)]. After sterilization the pH (5.3.2.6) of the medium shall be

equivalent to 7,2 ± 0,2.
5.2.2.4 Malt extract agar (MEA)
For maintenance of fungal strains, sporulation and performance of viable counts.
Malt extract [food grade (e.g. Christomalt powder from Difal) or 30,0 g
an equivalent extract that is not highly purified and not only
based on maltose (e.g. Malt extract from OXOID)]
Agar 15,0 g
Water (5.2.2.2) to 1 000,0 ml
However, if there are problems producing at least 75 % spiny spores see 5.4.1.3.

Sterilize in the autoclave [5.3.2.1 a)]. After sterilization, the pH (5.3.2.6) of the medium shall be

equivalent to 5,6 ± 0,2.

In case of an encountering problems with neutralization (5.5.2.3 and 5.5.2.4), it may be necessary to add

neutralizer (5.2.2.6) to MEA (5.2.2.4). Annex B gives guidance on the neutralizers that can be used. It is

recommended not to use neutralizer that causes opalescence in the agar.
5.2.2.5 Diluent
Tryptone sodium chloride solution:
Tryptone, pancreatic digest of casein 1,0 g
Sodium chloride 8,5 g
Water (5.2.2.2) to 1 000,0 ml

This Malt extracts from Difal and OXOID are examples of a suitable product available commercially. This

information is given for the convenience of users of this European Standard and does not constitute an

endorsement by CEN of this product.
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SIST EN 17387:2022
EN 17387:2021 (E)

Sterilize in the autoclave [5.3.2.1 a)]. After sterilization, the pH (5.3.2.6) of the diluent shall be

equivalent to 7,0 ± 0,2.
5.2.2.6 Neutralizer

The neutralizer shall be validated for the product being tested in accordance with 5.5.2.3 and 5.5.2.4. It

shall be sterile.

NOTE Information on neutralizers that have been found to be suitable for some categories of products is

given in Annex B.
5.2.2.7 Hard water for dilution of products
For the preparation of 1 l of hard water, the procedure is as follows:

— Prepare solution A: dissolve 19,84 g magnesium chloride (MgCl ) and 46,24 g calcium chloride

(CaCl ) in water (5.2.2.2) and dilute to 1 000 ml. Sterilize by membrane filtration (5.3.2.19) or in the

autoclave [5.3.2.1 a)]. Autoclaving – if used - can cause a loss of liquid. In this case make up to

1 000 ml with water (5.2.2.2) under aseptic conditions. Store the solution in the refrigerator

(5.3.2.15) for no longer than one month.

— Prepare solution B: dissolve 35,02 g sodium bicarbonate (NaHCO ) in water (5.2.2.2) and dilute to

1000 ml. Sterilize by membrane filtration (5.3.2.19). Store the solution in the refrigerator (5.3.2.15)

for no longer than one week.

— Place 600 ml to 700 ml of water (5.2.2.2) in a 1 000 ml volumetric flask (5.3.2.13) and add 6,0 ml of

solution A, then 8,0 ml of solution B. Mix and dilute to 1 000 ml with water (5.2.2.2). The pH

(5.3.2.6) of the hard water shall be 7,0 ± 0,2. If necessary, adjust the pH by using a solution of

approximately 40 g/l (about 1 mol/l) of sodium hydroxide (NaOH) or approximately 36,5 g/l

(about 1 mol/l) of hydrochloric acid (HCl).

The hard water shall be freshly prepared under aseptic conditions and used within 12 h.

NOTE When preparing the product test solutions (5.4.2), the addition of the product to the hard water

produces different final water hardness in each test tube. In any case, the final hardness expressed as calcium

carbonate (CaCO ) is lower than 375 mg/l in the test tube.
5.2.2.8 Interfering substance
5.2.2.8.1 General

The interfering substance shall be chosen according to the conditions of use laid down for the product.

The interfering substance shall be sterile and prepared at 2 times its final concentration in the test.

For any additional interfering substance, the ionic composition (e.g. pH, calcium and/or magnesium

hardness) and chemical composition (e.g. mineral substances, protein, carbohydrates, lipids,

detergents) shall be fully defined.

NOTE The term “interfering substance” is used even if it contains more than one substance.

The methods of preparation and sterilization together with the composition shall be noted in the test

report (5.7).
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SIST EN 17387:2022
EN 17387:2021 (E)
5.2.2.8.2 Bovine albumin solution
Bovine albumin solutions for the test conditions shall be prepared as follows.
a) Preparation for clean conditions:

— Dissolve 0,3 g of bovine albumin fraction V (suitable for microbiological purposes) in 100 ml of

diluent (5.2.2.5).
— Sterilize by membrane filtration (5.3.2.19).
— Dilute to the 1/5th with diluent (5.2.2.5).
— Keep in the refrigerator and use within one month.

The final concentration of bovine albumin in the test procedure (5.5.2) is 0,3 g/l.

b) Preparation for dirty conditions:

— Dissolve 3,0 g of bovine albumin fraction V (suitable for microbiological purposes) in 97 ml of

diluent (5.2.2.5).
— Sterilize by membrane filtration (5.3.2.19).

— Prepare at least 8,0 ml fresh sterile defibrinated sheep blood. Centrifuge the sheep blood at 800

g (5.3.2.14) for 10 min. After discarding the supernatant, resuspend erythrocytes in diluent

(5.2.2.5).
— Repeat this procedure at least 3 times, until the supernatant is colourless.

— Resuspend 3 ml of the packed sheep erythrocytes in the 97 ml of sterilized bovine albumin

solution (above).
— Dilute to the 1/5th with diluent (5.2.2.5).

— To avoid contamination this mixture should be split in portions probably needed per day and

kept in separate containers for a maximum of 7 days in a refrigerator (5.3.2.15).

The final concentration of bovine albumin and sheep erythrocytes in the test procedure (5.5) shall be

3 g/l and 3 ml/l respectively.
5.2.3 Test surface

The test surface is a flat stainless steel 304 ([1],[2]) disc (2 cm diameter) with grade 2b finish on both

sides. Only one side of a disc shall be used. The discs shall be used only once and manipulated with

forceps (5.3.2.16).

For cleaning, the discs shall be placed in a beaker (minimum size: 50 ml) containing not less than 20 ml

of 5 % (V/V) Decon for 60 min. Immediately rinse the discs twice with water (5.2.2.2) for 10 s to

completely remove the cleanser. Do not let the discs dry.
3 ®

Decon is an example of a suitable product available commercially. This information is given for the

convenience of users of this standard and does not constitute an endorsement by CEN of this product.

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SIST EN 17387:2022
EN 17387:2021 (E)

NOTE To supply a satisfactory flow of water, a sterilized fluid dispensing pressure vessel with suitable hose

and connectors or other suitable method can be used and regulated to supply approximately 2 000 ml per min.

For decontamination, place the clean disc in a bath containing 70 % (V/V) iso-propanol for 15 min.

Remove the disc and dry by evaporation under laminar air flow (5.3.2.17).
5.3 Apparatus and glassware
5.3.1 General

Sterilize all glassware and parts of the apparatus that will come into contact with the culture media and

reagents or the sample, except those which are supplied sterile, by one of the following methods:

a) by moist heat, in the autoclave [5.3.2.1 a)];
b) by dry heat, in the hot air oven [5.3.2.1 b)].
5.3.2 Usual microbiological laboratory equipment and, in partic
...

SLOVENSKI STANDARD
oSIST prEN 17387:2019
01-julij-2019
Kemična razkužila in antiseptiki - Kvantitativni preskus na neporoznih površinah
za vrednotenje baktericidnega in/ali fungicidnega delovanja in/ali delovanja
kemičnih razkužil na kvasovke v humani medicini - Preskusna metoda in zahteve
brez mehanskega delovanja (faza 2, stopnja 2)

Chemical disinfectants and antiseptics - Quantitative non-porous surface test for the

evaluation of bactericidal and/or yeasticidal and/or fungicidal activity of chemical

disinfectants used in medical area - Test method and requirements without mechanical

action (phase 2, step 2)

Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflächen‐Versuch zur

Bestimmung der bakteriziden und/oder levuroziden und/oder fungiziden Wirkung

chemischer Desinfektionsmittel auf nicht porösen Oberflächen im humanmedizinischen

Bereich - Prüfverfahren und Anforderungen ohne mechanische Behandlung (Phase 2,
Stufe 2)
Ta slovenski standard je istoveten z: prEN 17387
ICS:
11.080.20 Dezinfektanti in antiseptiki Disinfectants and antiseptics
oSIST prEN 17387:2019 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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oSIST prEN 17387:2019
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oSIST prEN 17387:2019
DRAFT
EUROPEAN STANDARD
prEN 17387
NORME EUROPÉENNE
EUROPÄISCHE NORM
May 2019
ICS 11.080.20; 71.100.35
English Version
Chemical disinfectants and antiseptics - Quantitative non-
porous surface test for the evaluation of bactericidal
and/or yeasticidal and/or fungicidal activity of chemical
disinfectants used in medical area - Part rev: Test method
and requirements without mechanical action (phase 2,
step 2)
Chemische Desinfektionsmittel und Antiseptika -
Quantitativer Oberflächen-Versuch zur Bestimmung
der bakteriziden und/oder levuroziden und/oder
fungiziden Wirkung chemischer Desinfektionsmittel
auf nicht porösen Oberflächen im humanmedizinischen
Bereich - Prüfverfahren und Anforderungen ohne
mechanische Behandlung (Phase 2, Stufe 2)

This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee

CEN/TC 216.

If this draft becomes a European Standard, 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.

This draft European Standard was established by CEN 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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,

Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

Turkey and United Kingdom.

Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are

aware and to provide supporting documentation.

Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without

notice and shall not be referred to as a European Standard.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels

© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 17387:2019 E

worldwide for CEN national Members.
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oSIST prEN 17387:2019
prEN 13787:2019 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms and definitions ................................................................................................................................... 5

4 Requirements ................................................................................................................................................... 6

5 Test methods .................................................................................................................................................... 6

5.1 Principle ............................................................................................................................................................. 6

5.2 Materials and reagents .................................................................................................................................. 7

5.2.1 Test organisms ................................................................................................................................................. 7

5.2.2 Culture media and reagents ........................................................................................................................ 7

5.2.3 Test surface .................................................................................................................................................... 10

5.3 Apparatus and glassware .......................................................................................................................... 11

5.3.1 General ............................................................................................................................................................. 11

5.3.2 Usual microbiological laboratory equipment and, in particular, the following: .................. 11

5.4 Preparation of test organism suspensions and product test solutions .................................... 12

5.4.1 Test organism suspensions ...................................................................................................................... 12

5.4.2 Product test solutions ................................................................................................................................. 18

5.5 Procedure for assessing the bactericidal, yeasticidal and fungicidal activity of the

product ............................................................................................................................................................. 19

5.5.1 Choice of experimental conditions ........................................................................................................ 19

5.5.2 Test procedure .............................................................................................................................................. 19

5.5.3 Counting of the test mixtures ................................................................................................................... 23

5.6 Calculation and expression of results ................................................................................................... 23

5.6.1 Calculation of data ....................................................................................................................................... 23

5.6.2 Verification of methodology ..................................................................................................................... 25

5.6.3 Expression of results ................................................................................................................................... 25

5.6.4 Conclusion....................................................................................................................................................... 25

5.7 Test report ...................................................................................................................................................... 26

Annex A (informative) Corresponding referenced strains ........................................................................ 28

Annex B (informative) Neutralizers ................................................................................................................... 29

Annex C (informative) Expression of results with the dilution-neutralization method ................. 31

Annex D (informative) Bactericidal activity on surfaces for general purposes under clean

conditions ....................................................................................................................................................... 33

Annex E (informative) Vacuum drying by desiccator - Instruction of use ............................................ 35

E.1 Description ..................................................................................................................................................... 35

E.2 Procedure........................................................................................................................................................ 35

E.3 Other types of desiccators ......................................................................................................................... 36

Annex F (informative) Precision of the test result ........................................................................................ 37

Bibliography ................................................................................................................................................................. 38

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European foreword

This document (prEN 13787:2019) has been prepared by Technical Committee CEN/TC 216 “Chemical

disinfectants and antiseptics”, the secretariat of which is held by AFNOR.
This document is currently submitted to the CEN Enquiry.
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Introduction

This document describes a surface test method for establishing whether a chemical disinfectant in the

area and fields described in the scope (Clause 1) has or does not have bactericidal and/or fungicidal or

yeasticidal activity on non-porous surfaces without mechanical action.

The laboratory test closely simulates practical conditions of application. Chosen conditions (contact time,

temperature, and microorganisms on surfaces...) reflect parameters which are found in practical

situations including conditions which may influence the action of disinfectants.

The conditions are intended to cover general purposes and to allow reference between laboratories and

product types.

However, for some applications the recommendations of use of a product can differ and therefore

additional test conditions need to be used.
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1 Scope

This document specifies a test method and the minimum requirements for bactericidal and yeasticidal

and additionally fungicidal activity of chemical disinfectant products that form a homogeneous,

physically stable preparation when diluted with hard water – or in the case of ready-to-use products –

with water.

NOTE 1 Dilutions are necessary as three concentrations in the active to non-active range are tested.

This document applies to products that are used in the medical area for disinfecting non-porous surfaces

without mechanical action.

This document applies to areas and situations where disinfection or antisepsis is medically indicated.

Such indications occur in patient care, for example:
— in hospitals, in community medical facilities and in dental institutions;
— in clinics of schools, of kindergartens and of nursing homes;

and can occur in the workplace and in the home. It can also include services such as laundries and

kitchens supplying products directly for the patients.

EN 14885 specifies in detail the relationship of the various tests to one another and to use

recommendations.

Using this document, it is possible to determine the activity of products like commercial formulations or

active substances on bacteria and/or fungi in the conditions in which they are used and therefore it

corresponds to a phase 2, step 2 test.

This method excludes the evaluation of the activity of products against mycobacteria and bacteria spores.

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.

EN 12353, Chemical disinfectants and antiseptics - Preservation of test organisms used for the

determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal

(including bacteriophages) activity

EN 14885, Chemical disinfectants and antiseptics - Application of European Standards for chemical

disinfectants and antiseptics
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN 14885.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at http://www.iso.org/obp
— IEC Electropedia: available at http://www.electropedia.org/
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4 Requirements

The product shall demonstrate at least a 5 decimal log (lg) reduction for bacteria and at least a 4 decimal

log (lg) reduction for fungi, when tested in accordance with Table 1 and 5.5.
Table 1 — Experimental conditions
Test conditions Bactericidal activity Yeasticidal activity Fungicidal activity
Staphylococcus aureus
Minimum Aspergillus
spectrum of Enterococcus hirae Candida albicans brasiliensis
test organisms Candida albicans
Pseudomonas aeruginosa
additional any relevant test organism
according to the manufacturer’s recommendation, but between
Test
4 °C ± 1°C to 30 °C ± 1°C
temperature
For tests performed at room temperature, the range shall be 20 °C ± 2,5°C
according to the manufacturer’s recommendation, but at minimum 1 min
Contact time a
and no longer than 5 min or 60 min (from 1 min to 5 min at intervals of 1 min
and from 5 min to 60 min at intervals of 5 min)
Interfering substance
clean conditions 0,3 g/l bovine albumin
dirty conditions 3,0 g/l bovine albumin plus 3,0 ml erythrocytes
additional any relevant substance

The contact times for surface disinfectants stated in this table are chosen on the basis of the practical

conditions of the product. The recommended contact time for the use of the product is within the

responsibility of the manufacturer. Products intended to disinfect surfaces that are likely to come into

contact with the patient and / or the medical staff and surfaces, which are frequently touched by different

people, leading to the transmission of microorganisms to the patient, shall be tested with a contact time of

maximum 5 min. The same applies where the contact time of the product shall be limited for practical

reasons. Products for other surfaces than stated above may be tested with a contact time of maximum 60 min

NOTE For the additional conditions, the concentration defined as a result can be lower than the one

obtained under the minimum test conditions.
5 Test methods
5.1 Principle

A test suspension of bacteria or fungi in a solution of interfering substances is inoculated onto a test

stainless steel surface and dried. A sample of the product under test is applied in a manner that

completely covers the dried test organisms. The surface is maintained at the temperature and contact

time exemplified in Clause 4 and 5.5.1. At the end of the contact time the surface is transferred to a

previously validated neutralization medium to suppress any product activity. The number of surviving

test organisms which can be recovered from the surface is determined quantitatively.

The number of bacteria or fungi on a test surface treated with hard water in place of the product under

test is also determined and the reduction in viable counts attributed to the product is calculated as the

difference between the two test surfaces’ results.
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5.2 Materials and reagents
5.2.1 Test organisms
The bactericidal activity shall be evaluated using the following strains :
— Staphylococcus aureus ATCC 6 538;
— Enterococcus hirae ATCC 10 541;
— Pseudomonas aeruginosa ATCC 15 442.
The fungicidal activity shall be evaluated using the following strains :
— Aspergillus brasiliensis ATCC 16 404.
— Candida albicans ATCC 10 231;
The yeasticidal activity shall be evaluated using the following strain :
— Candida albicans ATCC 10 231.
If required for specific applications, additional strains may be chosen.

NOTE See Annex A for corresponding strain numbers in some other culture collections.

If additional test organisms are used, they shall be incubated under optimum growth conditions

(temperature, time, atmosphere, media) noted in the test report. If the additional test organisms selected

do not correspond to the specified strains, their suitability for supplying the required inocula shall be

verified. If these additional test organisms are not classified at a reference centre, their identification

characteristics shall be stated. In addition, they shall be held by the testing laboratory or national culture

collection under a reference for five years.
5.2.2 Culture media and reagents
5.2.2.1 General

All weights of chemical substances given in this European Standard refer to the anhydrous salts. Hydrated

forms may be used as an alternative, but the weights required shall be adjusted to allow for consequent

molecular weight differences.

The reagents shall be of analytical grade and/or appropriate for microbiological purposes. They shall be

free from substances that are toxic or inhibitory to the test organisms.

To improve reproducibility, it is recommended that commercially available dehydrated material is used

for the preparation of culture media. The manufacturer's instructions relating to the preparation of these

products should be rigorously followed.
For each culture medium and reagent, a time limitation for use should be fixed.
All specified pH values are measured at (20 ± 1) °C.

The ATCC numbers are the collection numbers of strains supplied by these culture collections. This information

is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of

the product named
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5.2.2.2 Water

The water shall be freshly glass-distilled water and not demineralized water. If distilled water of adequate

quality is not available, water for injections (see bibliographic reference [1]) may be used.

Sterilize in the autoclave [5.3.2.1a)]. Sterilization is not necessary if the water is used e.g. for preparation

of culture media and subsequently sterilized.
NOTE See 5.2.2.7 for the procedure to prepare hard water.
5.2.2.3 Tryptone Soya Agar (TSA)
For maintenance of bacterial strains and performance of viable counts.
Tryptone, pancreatic digest of casein 15,0 g
Soya peptone, papaic digest of Soybean meal 5,0 g
Sodium chloride (NaCl) 5,0 g
Agar 15,0 g
Water (5.2.2.2) to 1 000,0 ml

Sterilize in the autoclave [5.3.2.1 a)]. After sterilization the pH (5.3.2.6) of the medium shall be equivalent

to 7,2 ± 0,2.
5.2.2.4 Malt extract agar (MEA)
For maintenance of fungal strains, sporulation and performance of viable counts.
Malt extract [food grade (e.g. Christomalt powder from Difal) or 30,0 g
an equivalent extract that is not highly purified and not only
based on maltose (e.g. Malt extract from OXOID)]
Agar 15,0 g
Water (5.2.2.2) to 1 000,0 ml
However, if there are problems producing at least 75 % spiny spores see 5.4.1.3.

Sterilize in the autoclave [5.3.2.1 a)]. After sterilization, the pH (5.3.2.6) of the medium shall be equivalent

to 5,6 ± 0,2.

In case of an encountering problems with neutralization (5.5.2.3 and 5.5.2.4), it may be necessary to add

neutralizer to MEA. Annex B gives guidance on the neutralizers that may be used. It is recommended not

to use neutralizer that causes opalescence in the agar.
5.2.2.5 Diluent
Tryptone sodium chloride solution:
Tryptone, pancreatic digest of casein 1,0 g
Sodium chloride 8,5 g
Water (5.2.2.2) to 1 000,0 ml

This Malt extracts from Difal and OXOID are examples of a suitable product available commercially. This

information is given for the convenience of users of this European Standard and does not constitute an endorsement

by CEN of this product
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Sterilize in the autoclave [5.3.2.1 a)]. After sterilization, the pH (5.3.2.6) of the diluent shall be equivalent

to 7,0 ± 0,2.
5.2.2.6 Neutralizer

The neutralizer shall be validated for the product being tested in accordance with 5.5.2.3 and 5.5.2.4. It

shall be sterile.

NOTE Information on neutralizers that have been found to be suitable for some categories of products is given

in Annex B.
5.2.2.7 Hard water for dilution of products
For the preparation of 1 l of hard water, the procedure is as follows:

— prepare solution A: dissolve 19,84 g magnesium chloride (MgCl ) and 46,24 g calcium chloride

(CaCl ) in water (5.2.2.2) and dilute to 1 000 ml. Sterilize by membrane filtration (5.3.2.19) or in the

autoclave [5.3.2.1 a)]. Autoclaving – if used - may cause a loss of liquid. In this case make up to

1 000 ml with water (5.2.2.2) under aseptic conditions. Store the solution in the refrigerator

(5.3.2.15) for no longer than one month;

— prepare solution B: dissolve 35,02 g sodium bicarbonate (NaHCO ) in water (5.2.2.2) and dilute to

1000 ml. Sterilize by membrane filtration (5.3.2.19). Store the solution in the refrigerator (5.3.2.15)

for no longer than one week;

— place 600 ml to 700 ml of water (5.2.2.2) in a 1 000 ml volumetric flask (5.3.2.13) and add 6,0 ml of

solution A, then 8,0 ml of solution B. Mix and dilute to 1 000 ml with water (5.2.2.2). The pH (5.3.2.6)

of the hard water shall be 7,0 ± 0,2. If necessary, adjust the pH by using a solution of approximately

40 g/l (about 1 mol/l) of sodium hydroxide (NaOH) or approximately 36,5 g/l (about 1 mol/l) of

hydrochloric acid (HCl).

The hard water shall be freshly prepared under aseptic conditions and used within 12 h.

NOTE When preparing the product test solutions (5.4.2), the addition of the product to the hard water

produces different final water hardness in each test tube. In any case, the final hardness expressed as calcium

carbonate (CaCO ) is lower than 375 mg/l in the test tube.
5.2.2.8 Interfering substance
5.2.2.8.1 General

The interfering substance shall be chosen according to the conditions of use laid down for the product.

The interfering substance shall be sterile and prepared at 2 times its final concentration in the test.

For any additional interfering substance, the ionic composition (e.g. pH, calcium and/or magnesium

hardness) and chemical composition (e.g. mineral substances, protein, carbohydrates, lipids, detergents)

shall be fully defined.

NOTE The term “interfering substance” is used even if it contains more than one substance.

The method of preparation and sterilization together with the composition shall be noted in the test

report (5.7).
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5.2.2.8.2 Bovine albumin solution
Bovine albumin solutions for the test conditions shall be prepared as follows.
a) Preparation for clean conditions

— dissolve 0,3 g of bovine albumin (Cohn fraction V for Dubos medium) in 100 ml of diluent (5.2.2.5)

— sterilize by membrane filtration (5.3.2.19)
— Dilute to the 1/5th with diluent (5.2.2.5)
— Keep in the refrigerator and use within one month

The final concentration of bovine albumin in the test procedure (5.5.2) is 0,3 g/l.

b) Preparation for dirty conditions

— Dissolve 3,00 g of bovine albumin fraction V (suitable for microbiological purposes) in 97 ml of

diluent (5.2.2.5)
— Sterilize by membrane filtration (5.3.2.19)

— Prepare at least 8,0 ml fresh sterile defibrinated sheep blood. Centrifuge the sheep blood at 800 g

(5.3.2.14) for 10 min. After discarding the supernatant, resuspend erythrocytes in diluent (5.2.2.5)

— Repeat this procedure at least 3 times, until the supernatant is colourless

— Resuspend 3 ml of the packed sheep erythrocytes in the 97 ml of sterilized bovine albumin solution

(above)
— Dilute to the 1/5th with diluent (5.2.2.5)

— To avoid contamination this mixture should be split in portions probably needed per day and kept in

separate containers for a maximum of 7 days in a refrigerator (5.3.2.15)

The final concentration of bovine albumin and sheep erythrocytes in the test procedure (5.5) shall be

3 g/l and 3 ml/l respectively.
5.2.3 Test surface

The test surface is a flat stainless steel 304 disc (2 cm diameter) with grade 2b finish on both sides. Only

one side of a disc shall be used. The discs shall be used only once.

For cleaning, the discs shall be placed in a beaker (minimum size: 50 ml) containing not less than 20 ml

of 5 % (V/V) Decon for 60 min. Immediately rinse the discs twice with water (5.2.2.2) for 10 s to

completely remove the cleanser. Do not let the discs dry.

NOTE To supply a satisfactory flow of water, a sterilized fluid dispensing pressure vessel with suitable hose

and connectors or other suitable method can be used and regulated to supply approximately 2 000 ml per min.

3 ®

Decon is an example of a suitable product available commercially. This information is given for the

convenience of users of this standard and does not constitute an endorsement by CEN of this product.

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prEN 13787:2019 (E)

For decontamination, place the clean disc in a bath containing 70 % (V/V) iso-propanol for 15 min.

Remove the disc and dry by evaporation under laminar air flow.
5.3 Apparatus and glassware
5.3.1 General

Sterilize all glassware and parts of the apparatus that will come into contact with the culture media and

reagents or the sample, except those which are supplied sterile, by one of the following methods:

a) by moist heat, in the autoclave [5.3.2.1 a)];
b) by dry heat, in the hot air oven [5.3.2.1 b)].

5.3.2 Usual microbiological laboratory equipment and, in particular, the following:

5.3.2.1 Apparatus for sterilization (moist and dry heat)

a) For moist heat sterilization, an autoclave capable of being maintained at ( 121 ) °C for a minimum

holding time of 15 min;
180 ) °C for a minimum
b) for dry heat sterilization, a hot air oven capable of being maintained at (
+5 +5

holding time of 30 min, at ( 170 ) °C for a minimum holding time of 1 h or at ( 160 ) °C for a

0 0
minimum holding time of 2 h.

5.3.2.2 Temperature controlled cabinet or chamber capable of being controlled at test temperatures

θ ± 1 °C (for test temperatures others than ambient).

5.3.2.3 Water baths, capable of being controlled at (20 ± 1) °C, at (45 ± 1) °C and at additional test

temperatures (θ ± 1) °C (5.5.1)

5.3.2.4 Incubator (for bactericidal activity), capable of being controlled either at (36 ± 1) °C or

(37 ± 1) °C. An incubator at (37 ± 1)°C may be used if an incubator at (36 ± 1)°C is not available. The

same temperature shall be used for incubations performed during a test and its control and validation

5.3.2.5 Incubator (for fungicidal or yeasticidal activity), capable of being controlled at (30 ± 1) °C

5.3.2.6 pH-meter, having an inaccuracy of calibration of no more than ± 0,1 pH units at (20 ± 1) °C

5.3.2.7 Stopwatch
5.3.2.8 Shakers
® 5
a) Electromechanical agitator, i.e. Vortex mixer ;
b) Mechanical shaker
5.3.2.9 Containers: Test tubes, culture bottles or flasks of suitable capacity
Disposable sterile equipment is an acceptable alternative to reusable glassware.
5 ®

Vortex is an example of a suitable product available commercially. This information is given for the

convenience of users of this European Standard and does not constitute an endorsement by CEN of this product.

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5.3.2.10 Graduated pipettes of nominal capacities 10 ml, 1 ml, 0,1 ml, 0,05 ml and 0,001 ml, or

calibrated automatic pipettes
5.3.2.11 Petri
...

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