SIST EN 17658:2023
(Main)Chemical disinfectants and antiseptics - Chemical textile disinfection for the domestic area - Test method and requirements (phase 2, step 2)
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 Mindestleistungsanforderungen an die mikrobizide Wirksamkeit eines chemischen Produkts fest, dessen vorgesehene Verwendung ein Waschvorgang im häuslichen Bereich in einer Haushaltswaschmaschine bei niedrigen Temperaturen (≤ 40 °C) ist. Dieses Verfahren gilt nicht für bestimmte Arten von Wäschedesinfektionstechnologien, für die spezielle Geräte erforderlich sind (d. h. Wirkstoffe werden in situ mithilfe spezieller Geräte erzeugt). Dieses Verfahren ist nicht auf bestimmte Arten von Textilien, von Produkten oder auf bestimmte Schritte im Waschzyklus beschränkt.
Entsprechend der Definition einer Prüfung der Phase 2, Stufe 2 ermittelt dieses Dokument die Wirksamkeit im Prüflabor, wobei die Praxisbedingungen für die Anwendung eines chemischen Produkts simuliert werden.
Dieses Dokument kann nicht angewendet werden, wenn die Desinfektion medizinisch indiziert ist (medizinischer Bereich) oder in hygienisch sensiblen Bereichen, in denen eine professionelle Aufbereitung der Wäsche erforderlich ist (z. B. Lebensmittelbereich, Gesundheitswesen, Medizin- und Reinraumsektor, PSA und Arbeitskleidung). In solchen Fällen gelten EN 16616 und EN 14065.
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 de performance relatives à l’efficacité microbicide d’un produit chimique destiné à être utilisé dans un procédé de lavage dans un environnement domestique, dans des machines à laver à usage domestique à basses températures (≤ 40 °C). Ce mode opératoire ne s’applique 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 à l’aide de dispositifs spécifiques). Cette méthode ne se limite pas à certains types de textiles, types de produits ou étapes du cycle de lavage.
Selon la définition d’un essai de phase 2, étape 2, ce document établit l’efficacité lors d’un essai en laboratoire simulant les conditions pratiques d’utilisation d’un produit chimique.
Le présent document ne peut pas s’appliquer lorsque la désinfection est à usage médical (secteur médical) ou dans les domaines sensibles aux normes d’hygiène dans lesquels le retraitement professionnel du linge est requis (c’est-à-dire, industrie agroalimentaire, secteur des soins de santé, secteur médical et salles blanches, EPI et vêtements de travail). Dans ces cas, l’EN 16616 et l’EN 14065 s’appliqueront.
NOTE Cette méthode correspond à un essai de phase 2, étape 2 (voir l’EN 14885).
L’EN 14885 précise de manière détaillée la relation entre les différents essais et les « recommandations d’utilisation ».
Kemična razkužila in antiseptiki - Kemično razkuževanje tekstila za domačo uporabo - Preskusne metode in zahteve (faza 2, stopnja 2)
Ta evropski standard določa preskusno metodo in minimalne zahteve za mikrobicidno aktivnost kemičnega izdelka, kadar je namenjen razkuževanju onesnaženega tekstila v gospodinjskem perilu in preprečevanju prenosa mikroorganizmov v pralno vodo in drug tekstil. To vključuje tudi izdelke, ki so namenjeni povečanju higiene gospodinjskega perila in/ali preprečevanju estetskih težav (npr. neprijetne vonjave, kvarjenje). S to metodo je mogoče izmeriti prispevek biocidnega proizvoda k zmanjšanju števila mikrobov v onesnaženih oblačilih in pralni tekočini. Opisana metoda je laboratorijski preskus faze 2, korak 2, ki simulira pogoje uporabe proizvoda in je namenjena določanju mikrobicidne aktivnosti proizvoda (PT2- dezinfekcija tekstila, gospodinjska uporaba). Ta metoda ni omejena na nekatere vrste tekstila, saj upošteva realno najslabši možni primer (bombažni nosilec in polibombažni balast). Ta metoda ni omejena na nekatere obremenitvene zmogljivosti balasta, saj upošteva realno najslabši možni primer glede na tržne študije (razmerje med tekočino 1:5) Ta metoda ni omejena na nekatere vrste navodil za uporabo izdelka (samostojno, kot kombinirani izdelek ali samostojno, vendar v kombinaciji z drugimi pralnimi izdelki, kot so detergenti). Ta metoda ni omejena na določene stopnje pranja, v katerih se izdelek uporablja (predpranje, glavno pranje ali izpiranje), saj je začetna mikrobna obremenitev realno najslabši možni primer, ki zajema vse situacije, pogoji umazanije pa so prilagojeni glede na praktične pogoje uporabe Pri tej metodi se za končno točko šteje konec stopnje pranja, v kateri naj bi izdelek deloval. Ta metoda ni primerna za tehnologije dezinfekcije perila, ki zahtevajo posebne naprave (npr. aktivne snovi, pridobljene na mestu samem z elektrolizo). Ta metoda ni namenjena uporabi za dezinfekcijo perila na zdravstvenem področju (za to uporabo glej standard EN 16616) Vsa ta druga področja uporabe ne bodo zajeta, saj ta metoda ni namenjena uporabi: drugi uporabi, kjer je potrebna dezinfekcija tekstila (npr. predelava hrane, hoteli, delovna oblačila, npr. iz farmacevtske industrije, laboratoriji, območja z živili ali podobne institucije).
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 17658:2023
01-april-2023
Kemična razkužila in antiseptiki - Kemično razkuževanje 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: EN 17658:2022
ICS:
71.100.35 Kemikalije za dezinfekcijo v Chemicals for industrial and
industriji in doma domestic disinfection
purposes
SIST EN 17658:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 17658:2023
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SIST EN 17658:2023
EN 17658
EUROPEAN STANDARD
NORME EUROPÉENNE
September 2022
EUROPÄISCHE NORM
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 European Standard was approved by CEN on 15 August 2022.
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, Türkiye 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
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17658:2022 E
worldwide for CEN national Members.
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Contents Page
European foreword .4
Introduction .5
1 Scope .6
2 Normative references .6
3 Terms and definitions .6
4 Requirements .7
5 Test method .9
5.1 Principle .9
5.2 Materials and reagents .9
5.2.1 Test organisms .9
5.2.2 Culture media and reagents .9
5.3 Apparatus and glassware . 11
5.3.1 General. 11
5.4 Preparation of test organism suspensions . 14
5.4.1 Test organism suspensions (test suspension) . 14
5.5 Procedure for assessing the microbicidal activity of the product . 16
5.5.1 General. 16
5.5.2 Method . 17
5.6 Experimental data and calculation . 21
5.6.1 Explanation of terms and abbreviations . 21
5.6.2 Calculation . 21
5.7 Verification of methodology . 25
5.7.1 General. 25
5.7.2 Control of weighted mean counts . 25
5.7.3 Basic limits . 25
5.8 Expression of results and precision . 26
5.8.1 Reduction . 26
5.8.2 Repetitions . 26
5.9 Interpretation of results – conclusion . 27
5.9.1 General. 27
5.9.2 Microbicidal activity . 27
5.10 Test report . 30
Annex A (informative) Referenced strains in national collections . 32
Annex B (informative) Suitable neutralizers and rinsing liquids . 34
B.1 General. 34
B.2 Neutralizers . 34
B.3 Neutralizer added to the agar for counting . 35
Annex C (informative) Graphical representations of the test method . 36
Annex D (informative) Example of lab-scale devices . 38
Annex E (informative) Test report (example) . 39
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Annex F (informative) SBL2004 composition . 46
Annex G (informative) Stainers and markers validated . 47
Bibliography . 48
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EN 17658:2022 (E)
European foreword
This document (EN 17658:2022) 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 March 2023, and conflicting national standards shall
be withdrawn at the latest by March 2023.
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,
Türkiye and the United Kingdom.
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Introduction
In the last few years, the need of energy saving has led to decreasing washing temperature of
domestic laundry to ≤ 40 °C. This fact compromises 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 rinse cycle) 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 360° around a horizontal axis (Rotawash, Launderometer, Gyrowash, Linitester and Mathis
BFA have been validated in the Ring Trial). This tumbling device maintains optimal agitation
[constant 40 r/min (±2 r/min)] 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 organisms and interfering
substance (conditions which may influence the action of the product in practice). The manufacturer’s
instructions should be sufficient to allow the method in this document to be carried out fully [dosing,
washing phase (main wash, rinse cycle) temperature and washing time].
This test pretends to generate a common experimental framework in which products can be tested
to specify 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 performance requirements for the
microbicidal efficacy of a chemical product intended for use in a wash process in a domestic
environment, in a domestic wash equipment at low temperatures (≤40 °C). This procedure does not
apply to certain types of laundry disinfection technologies which require specific devices (i.e. active
substances generated in situ using specific devices). This method is not limited to certain types of
textiles, types of products or steps in the washing cycle.
According to a phase 2, step 2 test definition, this document establishes the efficacy in laboratory test
simulating practical use conditions of a chemical product.
This document cannot be applied when the disinfection is medical indicated (medical area) or in
hygiene-sensitive areas where professional reprocessing of laundry is required (i.e. food, healthcare,
medical and cleanroom sectors, PPE, and workwear). In those cases, EN 16616 and EN 14065 will
apply.
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 14885:2022, 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 https://www.electropedia.org/
— ISO Online browsing platform: available at https://www.iso.org/obp
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3.1
liquor ratio
ratio of the weight of dry textile in kilogram and volume of wash liquor expressed in litre (w/v)
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 to the disinfection process to obtain disinfected textile
3.4
domestic laundry disinfection
treatment of textile (e.g., clothing, kitchen cloths, bed sheet, 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
biocidal treatment to avoid malodour and aesthetic damage caused by microorganisms
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
1) Domestic laundry disinfection
at least a reduction of bacteria on contaminated carriers of 4 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 lg/carrier are to be detected in cross contamination
carriers, and no more than 1,15 lg/ml in wash water.
2) Biocidal treatment to avoid malodour and aesthetic damage caused by microorganisms
at least a reduction of bacteria on contaminated carriers of 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 1,70 lg/carrier are to be detected in cross contamination
carriers, and no more than 2 lg/ml in wash water.
b) Rinse cycle
1) Domestic laundry disinfection
at least a reduction of bacteria on contaminated carriers of 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
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reached. Further, no more than 1,54 lg/carrier are to be detected in cross contamination
carriers, and no more than 1,15 lg/ml in wash water.
2) Biocidal treatment to avoid malodour and aesthetic damage caused by microorganisms
at least a reduction of bacteria on contaminated carriers of 2 lg, and a reduction of Candida
albicans of 2 lg. If an additionally fungicidal activity is claimed a reduction of 2 lg shall be
reached. Further, no more than 1,70 lg/carrier are to be detected in cross contamination
carriers, and no more than 2 lg/ml in wash water.
Table 1 — Minimum and additional test conditions
Test Domestic laundry disinfection Biocidal treatment to avoid
parameters malodour and aesthetic damage
caused by microorganisms
Main wash Rinse cycle Main wash Rinse cycle
Test Escherichia coli, Enterococcus hirae, E. coli, E. hirae, P. aeruginosa, S. aureus,
organisms Pseudomans aeruginosa, Staphylococcus
C. albicans
aureus,
Candida albicans
Additionally Additional test organism(s): Additional test organism(s):
a a
- Dermatophytes - Dermatophytes
- Organisms associated with malodour
b
Test As recommended As recommended As recommended As recommended
temperature by the by the by the by the
manufacturer manufacturer manufacturer manufacturer
c c c c
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
c c c c
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 specify by the To specify by the To specify by the To specify by the
manufacturer manufacturer manufacturer manufacturer
a
See Annex A for example of dermatophytes. This information is given for the convenience of users of this
document and does not constitute an endorsement by CEN.
b
See Annex A for examples of microorganisms related with malodour. This information is given for the
convenience of users of this document and does not constitute an endorsement by CEN.
c
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.
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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 (bovine serum albumin). 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 rinse cycle can be simulated. At the end of the disinfection step,
the product effect will be stopped by transferring the carriers into neutralizer solution (5.2.2.7) and
by shaking at 1 000 r/min in a mixer (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 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
1
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.
1
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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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.
NOTE 2 If distilled water of adequate quality is not available, water for injections (according to European
Pharmacopoeia) [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
2
(CaCl ) in water (5.2.2.2) and dilute to 1 000 ml. Sterilize by membrane filtration (5.3.2.7) or in
2
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 (NaHCO ) in water (5.2.2.2) and dilute
3
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 (5.2.2.2) 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 (5.3.2.4). 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 (5.2.2.2) 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
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.
10
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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.
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 3 g/l BSA solution.
5.2.2.8.2 Dirty conditions
Prepare test suspensions with a 3 g/l BSA solution.
For main wash conditions 3,5 g SBL2004 (SBL 2004, wfk Testgewebe, Brüggen, Germany) swatches
containing approx. 1,2 g standard soil (WFK item number 2814003) will be needed for each test
canister. For SBL2004 composition see Annex F.
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)].
2
5.3.2 Usual microbiological laboratory equipment
and, in particular, the following:
2
Disposable sterile equipment is an acceptable alternative to reusable glassware.
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5.3.2.1 Apparatus for sterilization (moist and dry heat)
+3
a) for moist heat sterilization, an autoclave capable of being maintained at ( 121 ) °C for a
0
minimum contact time of 15 min [2];
+5
b) for dry heat sterilization, a hot air oven capable of being maintained at ( 180 ) °C for a minimum
0
+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 (5.2.2.4 to 5.2.2.5).
5.3.2.5 Stopwatch
5.3.2.6 Shakers
a) Electromechanical agitator, e.g. vortex mixer;
b) mixer (
...
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.
3
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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
a
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
7
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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
1)
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.
1)
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
2
(CaCl ) in water (5.2.2.2) and dilute to 1 000 ml. Sterilize by membrane filtration (5.3.2.7) or in
2
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)].
2)
5.3.2 Usual microbiological laboratory equipment
and, in particular, the following:
5.3.2.1 Apparatus for sterilization (moist and dry heat)
+3
a) for moist heat sterilization, an autoclave capable of being maintained at ( 121 ) °C for a
0
minimum contact time of 15 min [2];
+5
b) for dry heat sterilization, a hot air oven capable of being maintained at ( 180 ) °C for a minimum
0
+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
®
3)
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
2)
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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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 )
N
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)
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5.3.2.18 Cotton carriers, 1 cm : The carriers are prepared by using standard cotton fabric
2
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
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