oSIST prEN 17658:2021

Chemical disinfectants and antiseptics - Chemical textile disinfection for the domestic area - Test method and requirements (phase 2, step 2)

This European Standard specifies a test method and the minimum requirements for the microbicidal activity of a chemical product when its intended to disinfect contaminated textiles in domestic laundry and prevent the transmission of microorganisms into the wash water and other textiles. This includes the intention of increasing the hygiene of domestic laundry and/or avoiding aesthetical problems (e.g. bad odour, spoilage). This method is able to measure the contribution of the biocidal product to the microbial reduction in contaminated cloths and washing liquor. The method described is a phase 2, step 2 laboratory test that simulates the conditions of application of the product and it is intended to determine the microbicidal activity of a product (PT2- textile disinfection, domestic use). This method is not limited to certain types of textiles as it considers a realistic worst-case (Cotton carrier and Polycotton ballast load) This method is not limited to certain ballast load capacities as it considered a realistic worst-case according to market studies (liquor ratio 1:5) This method is not limited to certain types of product use instructions (alone, as a combined product or alone but in combination with other laundry products such as detergents) This method is not limited to certain washing steps in which the product is used (pre-wash, main wash or rinsing) as the initial microbial load is a realistic worst case to cover all situations and soiling conditions are adapted according practical conditions of use This method considers the endpoint to be the end of the washing step where the product is intended to act. This method is not suitable for laundry disinfection technologies which require specific devices (e.g. active substances generated in situ through electrolysis). This method is not intended to be applied to laundry disinfection in the medical area (refer to EN 16616 for that application) All these other areas of application will be uncovered as this method in not intended to be applied: other applications where disinfecting textile is necessary (e.g. food processing, hotels, workwear e.g. from the pharmaceutical industry, laboratories, foodstuff areas or similar institutions).

Chemische Desinfektionsmittel und Antiseptika ¿ Chemische Textildesinfektion für den häuslichen Bereich ¿ Prüfverfahren und Anforderungen (Phase 2, Stufe 2)

Dieses Dokument legt ein Prüfverfahren für und die Mindestanforderungen an die mikrobizide Wirkung eines chemischen Produkts fest, dessen vorgesehene Verwendung die chemische Behandlung von Textilien im häuslichen Bereich ist, um die Hygieneleistung von Wäschepflegeprodukten für den häuslichen Bereich in Haushaltswaschmaschinen bei niedrigen Temperaturen (≤ 40 °C) zu bewerten. Dieses Verfahren gilt nicht für bestimmte Arten von Wäschedesinfektionstechnologien, für die spezielle Geräte erforderlich sind (d. h. Wirkstoffe werden in situ durch den Einsatz spezieller Geräte erzeugt). Dieses Verfahren ist nicht auf bestimmte Arten von Textilien, von Produkten oder auf bestimmte Schritte im Waschzyklus beschränkt.
Dieses Dokument kann auch für Produkte angewendet werden, die zur chemischen Desinfektion von Textilien in den Bereichen Lebensmittel, Industrie und öffentliche Einrichtungen (z. B. Lebensmittelverarbeitung, Geschäfte, Sporträume, Büros, Hotels, Arbeitskleidung, Lebensmittelbereiche oder ähnliche Einrichtungen) verwendet werden, jedoch nicht, wenn die Desinfektion medizinisch indiziert ist (medizinischer Bereich).
ANMERKUNG   Dieses Verfahren entspricht einer Prüfung der Phase 2, Stufe 2 (siehe EN 14885).
EN 14885 legt im Einzelnen die Beziehungen der verschiedenen Prüfungen untereinander und zu den "Anwendungsempfehlungen" fest.

Antiseptiques et désinfectants chimiques - Désinfection chimique du textile pour le domaine domestique - Méthode d'essai et prescriptions (phase 2, étape 2)

Le présent document spécifie une méthode d’essai et les prescriptions minimales relatives à l’activité microbicide d’un produit chimique destiné à être utilisé pour le traitement chimique des textiles dans le domaine domestique, afin d’évaluer les conditions de propreté du linge domestique dans des machines à laver à usage domestique à basse température (≤ 40 °C). Ce mode opératoire ne s’appliquera pas à certains types de technologies de désinfection du linge qui nécessitent des dispositifs spécifiques (c’est-à-dire, des substances actives générées in situ par l’utilisation de dispositifs spécifiques). Cette méthode ne se limite pas à certains types de textiles, types de produits ou étapes du cycle de lavage.
Le présent document peut également s’appliquer aux produits utilisés pour la désinfection chimique des textiles dans les secteurs agroalimentaire, industriel et en collectivité (par exemple, industrie agroalimentaire, magasins, salles de sport, bureaux, usines de fabrication de vêtements de travail, usine de production de denrées alimentaires ou établissements similaires), mais pas lorsque la désinfection est à usage médical (secteur médical).
NOTE   Cette méthode correspond à un essai de phase 2, étape 2 (voir EN 14885).
L’EN 14885 précise de manière détaillée la relation entre les différents essais et les recommandations d’« utilisation ».

Kemična razkužila in antiseptiki - Kemična dezinfekcija tekstila za domačo uporabo - Preskusne metode in zahteve (faza 2, stopnja 2)

General Information

• Status: Not Published
• Public Enquiry End Date: 02-May-2021
• ICS: 71.100.35 - Chemicals for industrial and domestic disinfection purposes
• Technical Committee: KDS - Cosmetics, chemical disinfectants and surface active agents
• Current Stage: 4020 - Public enquire (PE) (Adopted Project)
• Start Date: 04-Feb-2021
• Due Date: 24-Jun-2021
• Completion Date: 31-May-2021
• Ref Project: prEN 17658 - Chemical disinfectants and antiseptics - Chemical textile disinfection for the domestic area - Test method and requirements (phase 2, step 2)

Standards Content (sample)

SLOVENSKI STANDARD
oSIST prEN 17658:2021
01-april-2021
Kemična razkužila in antiseptiki - Kemična dezinfekcija tekstila za domačo
uporabo - Preskusne metode in zahteve (faza 2, stopnja 2)

Chemical disinfectants and antiseptics - Chemical textile disinfection for the domestic

area - Test method and requirements (phase 2, step 2)

Chemische Desinfektionsmittel und Antiseptika ¿ Chemische Textildesinfektion für den

häuslichen Bereich ¿ Prüfverfahren und Anforderungen (Phase 2, Stufe 2)

Antiseptiques et désinfectants chimiques - Désinfection chimique du textile pour le

domaine domestique - Méthode d'essai et prescriptions (phase 2, étape 2)
Ta slovenski standard je istoveten z: prEN 17658
ICS:
71.100.35 Kemikalije za dezinfekcijo v Chemicals for industrial and
industriji in doma domestic disinfection
purposes
oSIST prEN 17658:2021 en,fr,de

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

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oSIST prEN 17658:2021
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oSIST prEN 17658:2021
DRAFT
EUROPEAN STANDARD
prEN 17658
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2021
ICS 71.100.35
English Version
Chemical disinfectants and antiseptics - Chemical textile
disinfection for the domestic area - Test method and
requirements (phase 2, step 2)

Antiseptiques et désinfectants chimiques - Désinfection Chemische Desinfektionsmittel und Antiseptika ¿

chimique du textile pour le domaine domestique - Chemische Textildesinfektion für den häuslichen

Méthode d'essai et prescriptions (phase 2, étape 2) Bereich ¿ Prüfverfahren und Anforderungen (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, 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.

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

© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 17658:2021 E

worldwide for CEN national Members.
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Contents Page

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

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

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

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

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

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

5 Test method .....................................................................................................................................7

Annex A (informative) Referenced strains in national collections ........................................ 30

Annex B (informative) Suitable neutralizers and rinsing liquids .......................................... 31

B.1 General............................................................................................................................................ 31

B.2 Neutralizers .................................................................................................................................. 31

B.3 Neutralizer added to the agar for counting ....................................................................... 32

Annex C (informative) Graphical representations of the test method ................................. 33

Annex D (informative) Example of lab-scale device specification ......................................... 35

Annex E (informative) Test report (example) ............................................................................... 36

Bibliography ............................................................................................................................................... 42

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European foreword

This document (prEN 17658:2021) 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

Domestic laundry disinfection has taken relevance in the last few years with the new social trends of

energy saving based on decreasing the washing temperature of domestic laundry to ≤ 40°C. This fact,

together with the social trend of speediness, makes domestic laundry washes to be shorter in time

which compromise the level of microorganisms in the laundry items after the washing process.

Moreover, the fashion trends on textile design and fibre technology provides cloth items that need to

be washed in special care conditions (cool water, short cycles, soft chemistry) in order to preserve

their properties but at the same time without compromising their hygienic level.

Chemistry plays an important role to provide good hygienic conditions to domestic laundry under

such described conditions.

This document is a phase 2 step 2 test, specifies a lab-scale methodology for establishing if a chemical

product used in any of the domestic laundry procedures (main wash and rinsing) have a microbicidal

activity (bactericidal and yeasticidal activity) on contaminated textiles, washing bath and, an effect

in avoiding cross contamination of microorganisms from contaminated textiles to non-contaminated

textiles.

This lab-scale methodology is carried out by using a tumbling device able to rotate an exposure

chamber through 360° vertical orbit (Rotawash, Launderometer, Gyrowash, Linitester and Mathis

BFA have been validated in the Ring Trial). This tumbling device maintains optimal agitation

(constant 40 rpm (±2 rpm)) and precise temperature for consistently reliable test results.

Microorganisms are inoculated on textile carriers that are introduced in an exposure chamber to

simulate practical conditions including contact time, temperature, test organism and interfering

substance (conditions which may influence the action of the product in practice). The manufacturers

instructions should be sufficient to allow the method in this document to be carried out fully (dosing,

washing phase (main wash, rinsing) temperature and washing time).

This test pretends to generate a common experimental framework in which products can be tested

to define their effective dosage for each chosen experimental condition. Instructions for use

generated from the results of this test are the responsibility of manufacturers of products.

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1 Scope

This document specifies a test method and the minimum requirements for the microbicidal activity

of a chemical product which intended use is the chemical treatment of textiles in domestic area in

order to evaluate the hygiene performance of domestic laundry products within domestic washing

machines at low temperatures (≤40°C). This procedure will not apply to certain types of laundry

disinfection technologies which require specific devices (i.e active substances generated in situ

through the use of specific devices). This method is not limited to certain types of textiles, types of

products or steps in the washing cycle.

This document can also apply to products that are used for chemical disinfection of textiles in food,

industrial and institutional areas (e.g. food processing, shops, sport rooms, offices, hotels, workwear,

foodstuff areas or similar institutions) but not when the disinfection is medical indicated (medical

area).
NOTE This method corresponds to a phase 2, step 2 test (see EN 14885).

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

recommendations”.
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 1276, Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of

bactericidal activity of chemical disinfectants and antiseptics used in food, industrial, domestic and

institutional areas - Test method and requirements (phase 2, step 1)

EN 1650, Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of

fungicidal or yeasticidal activity of chemical disinfectants and antiseptics used in food, industrial,

domestic and institutional areas - Test method and requirements (phase 2, step 1)

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 and the following

apply.

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

— IEC Electropedia: available at http://www.electropedia.org/
— ISO Online browsing platform: available at https://www.iso.org/obp
3.1
liquor ratio

ratio of the weight of dry textile in kilogram and volume of wash liquor in litre (w/v)

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3.2
disinfection process

process taking into account practical conditions of application of the product including contact time,

temperature, test organisms and interfering substances to disinfect the textile
3.3
treatment of contaminated textile

handling the textile according the disinfection process to obtain disinfected textile

3.4
domestic and non-medical laundry disinfection

treatment of textile (e.g. clothing, kitchen cloths, bed sheet house, tablecloth…) with chemical

products to inactivate microbial load, the purpose of this level of disinfection being to prevent the

transmission of laundry microbiota (cloth, washing machine, wash water) between contaminated

and non-contaminated textiles
3.5
aesthetic and malodour treatment

reducing the microorganisms to a level that prevents negative impact to the laundry; damage to

aesthetics (e.g. spotting, discolouration, staining) or accumulation of microbial contamination

causing mal odour
4 Requirements

The product shall demonstrate at least the following lg reduction when tested in accordance with

Table 1 and fulfil the basic limits in 5.7.3.
a) Main wash
a. Domestic and non-medical laundry disinfection

at least a reduction of bacteria on contaminated carriers of more than 4 ulg, and a reduction

of Candida albicans of 3 lg. If an additionally fungicidal activity is claimed a reduction of 3

ulg shall be reached. Further, no more than 1,54 cfu (lg/carrier) are to be detected in cross

contamination carriers, and no more than 1,15 cfu (lg/ml) in wash water.
b. Aesthetic and malodour treatment

at least a reduction of bacteria on contaminated carriers of more than 3 lg, and a reduction

of Candida albicans of 3 ulg. If an additionally fungicidal activity is claimed a reduction of 3

lg shall be reached. Further, no more than 1,70 cfu (lg/carrier) are to be detected in cross

contamination carriers, and no more than 2 cfu (lg/ml) in wash water.
b) Rinsing cycle
a. Domestic and non-medical laundry disinfection

at least a reduction of bacteria on contaminated carriers of more than 3 lg, and a reduction

of Candida albicans of 3 lg. If an additionally fungicidal activity is claimed a reduction of 3 lg

shall be reached. Further, no more than 1,54 cfu (lg/carrier) are to be detected in cross

contamination carriers, and no more than 1,15 cfu (lg/ml) in wash water.
b. Aesthetic and malodour treatment
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at least a reduction of bacteria on contaminated carriers of more than 2 lg, and a reduction

of Candida albicans of 2 lg. If an additionally fungicidal activity is claimed a reduction of 3 lg

shall be reached. Further, no more than 1,70 cfu (lg/carrier) are to be detected in cross

contamination carriers, and no more than 2 cfu (lg/ml) in wash water.
Table 1 — Minimum and additional test conditions
Test Domestic and non-medical laundry Aesthetic and malodour treatment
parameters disinfection
Main wash Rinse cycle Main wash Rinse cycle

Test E. coli, E. hirae, P. aeruginosa, S. aureus, E. coli, E. hirae, P. aeruginosa, S. aureus,

organisms
C. albicans C. albicans
Additionally Additional test organism Additional test organism
Dermatophytes Dermatophytes
T. rubrum T. rubrum
T. mentagrophytes T. mentagrophytes
M. canis M. canis
Organisms associated with malodour Organisms associated with malodour
Test As recommended As recommended As recommended As recommended
temperature by the by the by the by the
manufacturer manufacturer manufacturer manufacturer
a a a a
and ≤ 40°C and ≤ 20°C and ≤ 40°C and ≤ 20°C
Contact time As recommended As recommended As recommended As recommended
by the by the by the by the
a a a a
manufacturer manufacturer manufacturer manufacturer
Soiling Dirty: 3,5 g textile Clean: BSA in Dirty: 3,5 g textile Clean: BSA in
SBL2004 embedding matrix SBL2004 embedding matrix
(100g ballast (100g ballast
fabric) fabric)
Liquor ratio Between 1:2 and Between 1:2 and Between 1:2 and Between 1:2 and
1:10 1:10 1:10 1:10
To define by the To define by the To define by the To define by the
manufacturer manufacturer manufacturer manufacturer

The temperature and the contact time shall be chosen on the basis of the practical conditions of the

product application and within the responsibility of the manufacturer.
5 Test method
5.1 Principle

Textile carriers made of cotton fabric (5.3.2.18) will be contaminated with test suspension of

microorganisms in BSA. After drying the carriers will be placed in a tumbling lab testing device

containing textile ballast load in which the domestic disinfection process is simulated. Main wash

cycle and/or rinsing cycle can be simulated. At the end of the disinfection step, the product effect will

be stopped by transferring the carriers into neutralizer solution (5.2.1.6) and by shaking at 1000 rpm

in a thermomixer (5.3.2.6) to extract the remaining microorganisms from the carriers. The amount

of recovered microorganisms will be determined and the reduction rate will be calculated compared

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to an untreated control. In addition, the transfer rate to uninoculated carriers and contamination of

the wash liquor will be determined as well.
5.2 Materials and reagents
5.2.1 Test organisms

The bactericidal activity shall be evaluated using the following strains as test organisms

Pseudomonas aeruginosa ATCC 15442
Escherichia coli ATCC 10536
Staphylococcus aureus ATCC 6538
Enterococcus hirae ATCC 10541
The yeasticidal activity shall be evaluated using the following test organism:
Candida albicans ATCC 10231
NOTE See Annex A for strain reference in some other culture collections.

The required incubation temperatures for these test organisms are (36 ± 1) °C or (37 ± 1) °C (5.3.2.3)

[C. albicans (30 ± 1) °C]. The same temperature shall be used for all incubations performed during a

test and its controls and validation.

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

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

selected do not correspond to the specified strains/species, 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 stored 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 growth of 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 limitation for use should be fixed.
5.2.2.2 Water used for preparation of media

The water shall be fresh distilled water and not just demineralized water. Sterilize in the autoclave

(5.3.2.1 a).

NOTE 1 Sterilization is not necessary if the water is used e.g. for preparation of culture media and

subsequently sterilized.

The ATCC numbers are the collection numbers of strains supplied by the American Type Culture

Collection (ATCC). This information is given for the convenience of users of this document and does not

constitute an endorsement by CEN of the product named.
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NOTE 2 If distilled water of adequate quality is not available, water for injections (see bibliographic

reference [1]) can be used.
5.2.2.3 Water of standardized hardness
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 (5.3.2.7) 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

(5.3.2.8) at (2 to 8) °C for no longer than one month.

— Prepare solution B: Dissolve 35,02 g sodium bicarbonate (NaHCO3) in water (5.2.2.2) and dilute

to 1 000 ml. Sterilize by membrane filtration (5.3.2.7). Store the solution in the refrigerator

(5.3.2.8) at (2 to 8) °C for no longer than one week.

— Place 600 ml to 700 ml of water (5.2.1.1) in a 1 000 ml volumetric flask (5.3.2.12) and add with

the use of a pipette (5.3.2.9) 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 of the hard water shall be 7,0 ± 0,2, when measured at

(20 ± 1) °C (5.2.3.4). 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.

5.2.2.4 Tryptone Soy Agar (TSA)
Tryptone, pancreatic digest of casein 15,0 g
Soy peptone, papaic digest of soybean meal 5,0 g
Sodium chloride (NaCl) 5,0 g
Agar 15,0 g
Water (5.2.1.1) to 1 000,0 ml

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

7,2 ± 0,2 when measured at (20 ± 1) °C.

In case of encountering problems with neutralization (5.5.1.2 and 5.5.1.3) it may be necessary to add

neutralizer to the TSA. Annex B gives guidance on the neutralizers that may be used.

5.2.2.5 Malt Extract Agar (MEA)
Malt extract 30,0 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 of the medium shall be equivalent to

5,6 ± 0,2 when measured at (20 ± 1) °C.
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5.2.2.6 Diluent
Tryptone, pancreatic digest of casein 1,0 g
Sodium chloride (NaCl) 8,5 g
Water (5.2.2.2) to 1 000,0 ml

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

7,0 ± 0,2 when measured at (20 ± 1) °C.
5.2.2.7 Neutralizer

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

given in Annex B.
5.2.2.8 Interfering substance
5.2.2.8.1 Clean conditions
Prepare test suspensions with a 0,3 % BSA (Bovine serum albumin) solution.
5.2.2.8.2 Dirty conditions
Prepare test suspensions with a 0,3 % BSA solution.

For main wash conditions add 3,5 g SBL2004 swatches containing approx. 1,2 g standard soil (WFK

item number 2814003).
SBL2004 % composition
Vegatable Oil (Olio Extra Vergine di Oliva) 18 % – 20 %
Synthetic Sebum (BEY) 18 % – 20 %
Kaoline 8 % – 10 %
Proteine (from egg white powder) 8 % – 10 %
Bleach Consuming Agent 8 % – 10 %
Starch (corn starch) 6 % – 8 %
Salt (NaCl) 6 % – 8 %
Mineral Oil 6 % – 8 %
Lanolin 6 % – 8 %
Emulgator (Uniperol ® dispersing agent,
BASF) 1 % – 3 %
Urea (synthetic) 1 % – 3 %
Quartz 1 % – 3 %
Calciumchloride 1 % – 3 %
Carbon Black / Soot < 1 %
Iron Oxide black < 1 %
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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 contact time of 15 min [2];

b) for dry heat sterilization, a hot air oven capable of being maintained at ( 180 ) °C for a minimum

+5 +5

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

0 0
minimum contact time of 2 h.

5.3.2.2 Water baths, capable of being controlled between (20 ± 1) °C, and (45 ± 1) °C.

5.3.2.3 Incubator, capable of being controlled either at (36 ± 1) °C or (37 ± 1) °C for bactericidal

activity, and (30 ± 1) °C for yeasticidal activity. The same temperature shall be used for incubations

performed during a test and its controls and validation.

5.3.2.4 pH-meter, having an inaccuracy of calibration of no more than ± 0,1 pH units at

(20 ± 1) °C.

A puncture electrode or a flat membrane electrode should be used for measuring the pH of the agar

media (5.2.2.5 to 5.2.2.9).
5.3.2.5 Stopwatch
5.3.2.6 Shakers
a) Electromechanical agitator, e.g. Vortex mixer
b) Thermomixer (at 1000 rpm).

5.3.2.7 Membrane filtration apparatus, constructed of a material compatible with the

substances to be filtered, with a filter holder of at least 50 ml volume, and suitable for use of filters of

diameter 47 mm to 50 mm and 0,45 µm pore size for sterilization of hard water (5.2.2.3).

5.3.2.8 Refrigerator, capable of being controlled at (2 to 8) °C
Disposable sterile equipment is an acceptable alternative to reusable glassware.
3) ®

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

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

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prEN 17658:2021 (E)

5.3.2.9 Graduated pipettes, of nominal capacities 10 ml, 1 ml, 0,1 ml, and 0,01ml or calibrated

automatic pipettes
5.3.2.10 Sterile syringes, of nominal capacities 90 ml, and 10 ml
5.3.2.11 Petri dishes, (plates) of size 90 mm to 100 mm
5.3.2.12 Volumetric flasks
5.3.2.13 Centrifuge (4 700 g )
5.3.2.14 Coned bottom screw cap tubes (contents of 50 ml, diameter: about 28 mm)

5.3.2.15 Polypropylene micro tubes (contents 2ml) for recovery of the test organisms from the

carriers.
5.3.2.16 Stainless steel sieve (the holes shall be smaller than 1 cm × 1 cm)
5.3.2.17 Metal beads (0,9 g approx; 0,6 cm approx)

5.3.2.18 Cotton carriers, 1 cm : The carriers are prepared by using standard cotton fabric

thoroughly cut into 1 cm pieces, cooked in double-distilled water three times and sterilized in the

autoclave [see 5.3.2.1 a].
2 2
Mass per unit area: (170 ± 10) g/m (real 160 g/m )
Fibrous material (warp and weft): cotton, double corded
Fibre length (warp and weft): at least 27 mm
Yarn linear density (warp and weft): (295 ± 10) dtex
Yarn twist (warp and weft): Z-twist (700 ± 25) t/m
Weave: plain weave 1
Threads per unit length: 270 threads/dm each
The cotton control cloth shall be bleached, unfinished and not brightened.

After three washes the maximum tensile strength, wet, in the warp should be (63 ± 5) daN.

NOTE Cotton proofed in accordance with DIN 53919 [3] fulfils the requirements.
5.3.2.19 Carriers staining

In order to recognize which microorganism has been inoculated in each carrier, each carrier shall be

stained with a different colour.

It shall be validated that the dye used is not toxic for any of the microorganisms to be tested.

For example, Brauns-Heitmann staining or Mara by Marabu textile marker, are validated for this

purpose.
5.3.2.20 Ballast load

Textile of poly cotton (65 % polyester / 35 % cotton) shall be used as ballast load.

5.3.2.20.1 Rinsing cycle
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oSIST prEN 17658:2021
prEN 17658:2021 (E)
Total Amount of ballast load: 100g poly cotton textile.

Cut 100g of ballast fabric in to 10cm x 10cm squares. Boil with sterile distilled water two times. Air

dry fabric until completely dry. Autoclave 100g of ballast into a container or inside the canister and

autoclave for 15 min at 121°C, ensure fabric is completely dry before use.
5.3.2.20.2 Main wash cycle

Total Amount of ballast load: 96,5 g poly cotton textile in addition to 3,5 g of SBL2004.

Cut 96,5 g of ballast fabric in to 10cm x 10cm squares. Boil with sterile distilled water two times. Air

dry fabric until completely dry. Autoclave 96,5g of ballast into a container or inside the canister and

autoclave for 15 min at 121 °C, ensure fabric is completely dry before use.

The ballast load for product testing should not be used more than once. For water control ballast can

be reused 3 times before discarding.

5.3.2.21 Tumbling device (e.g. Rotawash, Launderometer, Gyrowash, Linitester, Mathis BFA)

The tumbling device shall warranty the following specifications:
a) Tumbling device to rotate canister through 360° vertical orbit.
b) Diameter at 40 rpm to 60 rpm.
c) Bath/Air temperature 20 °C – 40 °C.
d) Accuracy ± 1 °C.
5.3.2.22 Exposure chamber, Metal canister (1000 ml)
5.3.2.23 Container, capable of holding 100g of ballast (autoclavable)
5.4 Preparation of test organism suspensions
5.4.1 Test organism suspensions (test suspension)
5.4.1.1 General
For each test organism a test suspension to perform the test shall be prepared.
5.4.1.2 Preservation and stock cultures of test organisms

The test organisms and their stock cultures shall be prepared and kept in accordance with EN 12353.

5.4.1.3 Working culture and test organisms
5.4.1.3.1 Bacteria
In order to prepare the working culture of the bacteria, subculture fro
...