Chemical disinfectants and antiseptics - Quantitative non-porous surface test for the evaluation of bactericidal and/or fungicidal activity of chemical disinfectants used in food, industrial, domestic and institutional areas - Test method and requirements without mechanical action (phase 2, step 2)

This European Standard specifies a test method (phase 2/step 2) and the minimum requirements for bactericidal and/or fungicidal or yeasticidal activity of chemical disinfectants that form a homogeneous physically stable preparation in hard water or – in the case of ready-to-use products – with water in food, industrial, domestic and institutional areas, excluding areas and situations where disinfection is medically indicated and excluding products used on living tissues.
The scope of this European Standard applies at least to the following:
a)   Processing, distribution and retailing of:
1)   Food of animal origin:
i)   milk and milk products;
ii)   meat and meat products;
iii)   fish, seafood and products;
iv)   eggs and egg products;
v)   animal feeds;
vi)   etc.
2)   Food of vegetable origin:
i)   beverages;
ii)   fruits, vegetables and derivatives (including sugar distillery);
iii)   flour, milling and backing;
iv)   animal feeds;
v)   etc.
b)   Institutional and domestic areas:
1)   catering establishments;
2)   public areas;
3)   public transports;
4)   schools;
5)   nurseries;
6)   shops;
7)   sports rooms;
8)   waste container (bins);
9)   hotels;
10)   dwellings;
11)   clinically non sensitive areas of hospitals;
12)   offices;
13)   etc.
c)   Other industrial areas:
1)   packaging material;
2)   biotechnology (yeast, proteins, enzymes...);
3)   pharmaceutical;
4)   cosmetics and toiletries;
5)   textiles;
6)   space industry, computer industry;
7)   etc.
Using this European Standard, it is possible to determine the bactericidal or fungicidal or yeasticidal activity of the undiluted product. As three concentrations are tested, in the active to non active range, dilution of the product is required and, therefore, the product forms a homogeneous stable preparation in hard water.
EN 14885 specifies in detail the relationship of the various tests to one another and to use recommendations.
NOTE 1   The method described is intended to determine the activity of commercial formulations or active substances on bacteria and/or fungi in the conditions in which they are used.
NOTE 2   This method cannot be used to evaluate the activity of products against mycobacteria.

Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflächen-Versuch zur Bestimmung der bakteriziden und/oder fungiziden Wirkung chemischer Desinfektionsmittel auf nicht porösen Oberflächen in den Bereichen Lebensmittel, Industrie, Haushalt und öffentliche Einrichtungen - Prüfverfahren und Anforderungen ohne mechanische Behandlung (Phase 2, Stufe 2)

Diese Europäische Norm legt ein Prüfverfahren (Phase 2/Stufe 2) und die Mindestanforderungen an die bakterizide und/oder fungizide oder levurozide Wirkung von chemischen Desinfektionsmitteln in den Bereichen Lebensmittel, Industrie, Haushalt und in öffentlichen Einrichtungen fest, die mit Wasser standardisierter Härte oder – im Fall von gebrauchsfertigen Produkten – mit Wasser eine homogene, physikalisch stabile Lösung ergeben. Ausgenommen davon sind Bereiche und Situationen mit medizinisch indizierter Desinfektion und Produkte, die auf lebenden Geweben benutzt werden.
Diese Europäische Norm gilt mindestens für den folgenden Anwendungsbereich:
a)   Verarbeitung, Vertrieb und Verkauf von:
1)   Lebensmitteln tierischer Herkunft:
i)   Milch und Milchprodukte;
ii)   Fleisch und Fleischprodukte;
iii)   Fisch, Meerestiere und daraus hergestellte Erzeugnisse;
iv)   Eier und Eiprodukte;
v)   Tiernahrung;
vi)   usw.
2)   Lebensmitteln pflanzlicher Herkunft:
i)   Getränke;
ii)   Früchte, Gemüse und daraus hergestellte Erzeugnisse (einschließlich Zucker, Destillations-produkte);
iii)   Mehl, gemahlene und gebackene Produkte;
iv)   Tiernahrung;
v)   usw.
b)   gewerbliche Einrichtungen und Haushaltsbereiche:
1)   Verpflegungseinrichtungen;
2)   öffentliche Bereiche;
3)   öffentliche Verkehrsmittel;
4)   Schulen;
5)   Kindergärten;
6)   Geschäfte;
7)   Sportstätten;
8)   Abfallbehälter (Mülltonnen);
9)   Hotels;
10)   Wohngebäude;
11)   klinisch nicht relevante Bereiche von Krankenhäusern;
12)   Büroräume;
13)   usw.
c)   andere industrielle Bereiche:
1)   Verpackungsmaterial;
2)   Biotechnologie (Hefe, Proteine, Enzyme usw.);
3)   Hersteller von Pharmazeutika;
4)   Hersteller von Kosmetika und Toilettenartikeln;
5)   Hersteller von Textilien;
6)   Weltraumforschungsindustrie und Computerindustrie;
7)   usw.
Bei Anwendung dieser Europäischen Norm ist die Bestimmung der bakteriziden oder fungiziden oder levuroziden Wirkung des unverdünnten Produkts möglich. Da vom aktiven bis zum nicht aktiven Bereich drei Konzentrationen geprüft werden, ist eine Verdünnung des Produkts erforderlich, deshalb bildet das Produkt eine homogene stabile Lösung mit Wasser standardisierter Härte.
In EN 14885 ist die Beziehung zwischen den einzelnen Prüfungen untereinander und zu den Anwen-dungsempfehlungen detailliert festgelegt.
ANMERKUNG 1   Das beschriebene Verfahren bestimmt die Wirksamkeit gewerblich hergestellter Zubereitungen oder anderer aktiver Substanzen auf Bakterien und/oder Pilzen unter Anwendungsbedingungen.
ANMERKUNG 2   Dieses Verfahren kann nicht zur Bestimmung der Wirkung der Produkte gegenüber Mykobakterien angewendet werden.

Antiseptiques et désinfectants chimiques - Essai quantitatif de surface non-poreuse pour l'évaluation de l'activité bactéricide et/ou fongicide des désinfectants chimiques utilisés dans le domaine de l'agro-alimentaire, dans l'industrie, dans les domaines

La présente Norme européenne décrit une méthode d’essai (phase 2, étape 2) et les prescriptions minimales relatives à l’activité bactéricide et/ou fongicide ou levuricide des désinfectants chimiques qui forment une préparation homogène, physiquement stable dans l’eau dure ou – dans le cas des produits prêts à l'emploi, avec de l'eau – et qui sont utilisés dans le domaine de l’agro-alimentaire, dans l’industrie, dans le domaine domestique et en collectivités, à l’exception des domaines et des situations où la désinfection est médicalement prescrite et à l’exclusion des produits utilisés sur les tissus vivants.
Le domaine d’application de la présente Norme européenne couvre au moins les secteurs suivants :
a)   la fabrication, distribution et vente au détail de :
1)   les produits alimentaires d’origine animale :
i)   lait et produits laitiers ;
ii)   viande et produits carnés ;
iii)   poisson, fruits de mer et leurs produits dérivés;
iv)   œufs et produits dérivés ;
v)   alimentation animale ;
vi)   etc.
2)   les produits alimentaires d’origine végétale :
i)   boissons ;
ii)   fruits, légumes et leurs dérivés (y compris le sucre, les produits de distillation) ;
iii)   farine, minoterie et boulangerie ;
iv)   alimentation animale
v)   etc.
b)   les domaines domestiques et en collectivité :
1)   établissements de restauration ;
2)   lieux publics ;
3)   transports publics ;
4)   écoles ;
5)   crèches ;
6)   magasins ;
7)   salles de sport ;
8)   conteneurs pour déchets (poubelles) ;
9)   hôtels ;
10)   locaux d’habitation ;
11)   zones cliniquement non sensibles des hôpitaux ;
12)   bureaux ;
13)   etc.
c)   autres secteurs industriels :
1)   les matériaux d'emballage ;
2)   la biotechnologie (levure, protéines, enzymes...) ;
3)   l'industrie pharmaceutique ;
4)   les cosmétiques et les produits d'hygiène corporelle ;
5)   les textiles ;
6)   l’industrie spatiale, le secteur informatique ;
7)   etc.
Il est possible d’utiliser la présente Norme européenne pour déterminer l’activité bactéricide ou fongicide ou levuricide des produits prêts à l’emploi. Trois concentrations, dont une active et l’autre non active, sont soumises à essai, une dilution du produit est nécessaire et ce dernier forme donc une préparation homogène et stable dans l’eau dure.
L'EN 14885 spécifie en détail la relation entre les différents essais et les « recommandations d'emploi ».
NOTE 1   La méthode décrite est destinée à déterminer l’efficacité des formulations commerciales ou des substances actives sur les bactéries et/ou les champignons et les levures dans les conditions dans lesquelles elles doivent être utilisées.
NOTE 2   Cette méthode ne peut pas être utilisée pour évaluer l'activité des produits vis-à-vis des mycobactéries.

Kemična razkužila in antiseptiki - Kvantitativni preskus na neporoznih površinah za vrednotenje baktericidnega in/ali fungicidnega delovanja kemičnih razkužil v živilski in drugih industrijah, gospodinjstvu in javnih ustanovah - Preskusna metoda in zahteve brez mehanskega delovanja (faza 2, stopnja 2) (vključno z dopolnilom A1)

Ta evropski standard določa preskusno metodo (faza 2/stopnja 2) in minimalne zahteve za baktericidno in/ali fungicidno delovanje ali delovanje kemičnih razkužil na kvasovke, ki tvorijo homogen, fizikalno stabilen pripravek v trdi vodi ali – pri izdelkih, ki so pripravljeni za uporabo – in se z vodo uporabljajo v živilski in drugih industrijah, gospodinjstvu in javnih ustanovah, izključeni pa so področja in primeri, ko obstajajo zdravstvene indikacije za dezinfekcijo ter izdelki, ki se uporabljajo na živih tkivih.
Obseg tega evropskega standarda se uporablja vsaj za naslednje:
a) predelava, distribucija in prodaja:
1) hrane živalskega izvora:
i) mleko in mlečni izdelki;
ii) meso in mesni izdelki;
iii) ribe, morski sadeži in izdelki;
iv) jajca in jajčni izdelki;
v) živalska krma;
vi) itd.
2) hrane rastlinskega izvora:
i) pijače;
ii) sadje, zelenjava in derivati (vključno z destilati sladkorja);
iii) moka, mletje in peka;
iv) živalska krma;
v) itd.
b) javne ustanove in gospodinjstva:
1) priprava in dostava hrane;
2) javne ustanove;
3) javni prevozi;
4) šole;
5) vrtci;
6) trgovine;
7) športne dvorane;
8) zbiralniki odpadkov (koši);
9) hoteli;
10) bivališča;
11) klinično neobčutljivi prostori bolnišnic;
12) pisarne;
13) itd.
c) druga industrijska področja:
1) embalaža;
2) biotehnologija (kvasovke, proteini, encimi ipd.);
3) farmacevtski izdelki;
4) kozmetični in toaletni izdelki;
5) tkanine;
6) vesoljska in računalniška industrija;
7) itd.
S tem evropskim standardom je mogoče določiti baktericidno ali fungicidno delovanje ali delovanje kemičnih razkužil na kvasovke pri nerazredčenem izdelku. Ker so preskušene tri koncentracije od aktivnega do neaktivnega razpona, je potrebno razredčevanje izdelka in zato tvorijo homogen in stabilen pripravek v trdi vodi.
Standard EN 14885 podrobno določa razmerje med različnimi preskusi in priporočili za uporabo.
OPOMBA 1: Opisana metoda je namenjena določevanju dejavnosti komercialnih oblik ali aktivnih snovi na bakterijah in/ali glivah pod pogoji, v katerih se uporabljajo.
OPOMBA 2: Te metode ni mogoče uporabiti za vrednotenje delovanja izdelkov proti mikobakterijam.

General Information

Status
Published
Publication Date
30-Jul-2019
Current Stage
9000 - Start of review/2 Year Review Enquiry - Review Enquiry
Due Date
01-Apr-2020
Completion Date
01-Apr-2020

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SLOVENSKI STANDARD
SIST EN 13697:2015+A1:2019
01-november-2019
Nadomešča:
SIST EN 13697:2015
Kemična razkužila in antiseptiki - Kvantitativni preskus na neporoznih površinah
za vrednotenje baktericidnega in/ali fungicidnega delovanja kemičnih razkužil v
živilski in drugih industrijah, gospodinjstvu in javnih ustanovah - Preskusna
metoda in zahteve brez mehanskega delovanja (faza 2, stopnja 2) (vključno z
dopolnilom A1)

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

evaluation of bactericidal and/or fungicidal activity of chemical disinfectants used in food,

industrial, domestic and institutional areas - Test method and requirements without

mechanical action (phase 2, step 2)

Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflächen-Versuch zur

Bestimmung der bakteriziden und/oder fungiziden Wirkung chemischer
Desinfektionsmittel auf nicht porösen Oberflächen in den Bereichen Lebensmittel,

Industrie, Haushalt und öffentliche Einrichtungen - Prüfverfahren und Anforderungen

ohne mechanische Behandlung (Phase 2, Stufe 2)

Antiseptiques et désinfectants chimiques - Essai quantitatif de surface non-poreuse pour

l'évaluation de l'activité bactéricide et/ou fongicide des désinfectants chimiques utilisés

dans le domaine de l'agro-alimentaire, dans l'industrie, dans les domaines
Ta slovenski standard je istoveten z: EN 13697:2015+A1:2019
ICS:
71.100.35 Kemikalije za dezinfekcijo v Chemicals for industrial and
industriji in doma domestic disinfection
purposes
SIST EN 13697:2015+A1:2019 en,fr,de

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

---------------------- Page: 1 ----------------------
SIST EN 13697:2015+A1:2019
---------------------- Page: 2 ----------------------
SIST EN 13697:2015+A1:2019
EN 13697:2015+A1
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2019
EUROPÄISCHE NORM
ICS 11.080.20; 71.100.35 Supersedes EN 13697:2015
English Version
Chemical disinfectants and antiseptics - Quantitative non-
porous surface test for the evaluation of bactericidal
and/or fungicidal activity of chemical disinfectants used in
food, industrial, domestic and institutional areas - Test
method and requirements without mechanical action
(phase 2, step 2)

Antiseptiques et désinfectants chimiques - Essai Chemische Desinfektionsmittel und Antiseptika -

quantitatif de surface non-poreuse pour l'évaluation de Quantitativer Oberflächen-Versuch zur Bestimmung

l'activité bactéricide et/ou fongicide des désinfectants der bakteriziden und/oder fungiziden Wirkung

chimiques utilisés dans le domaine de l'agro- chemischer Desinfektionsmittel auf nicht porösen

alimentaire, dans l'industrie, dans les domaines Oberflächen in den Bereichen Lebensmittel, Industrie,

domestiques et en collectivité - Méthode d'essai sans Haushalt und öffentliche Einrichtungen - Prüfverfahren

action mécanique et prescriptions (phase 2/étape 2) und Anforderungen ohne mechanische Behandlung

(Phase 2, Stufe 2)

This European Standard was approved by CEN on 20 January 2015 and includes Amendment 1 approved by CEN on 10 June

2019.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this

European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references

concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN

member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by

translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management

Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,

Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,

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

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

© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 13697:2015+A1:2019 E

worldwide for CEN national Members.
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SIST EN 13697:2015+A1:2019
EN 13697:2015+A1:2019 (E)
Contents Page

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

Introduction .................................................................................................................................................................... 5

1 Scope .................................................................................................................................................................... 6

2 Normative references .................................................................................................................................... 7

3 Terms and definitions ................................................................................................................................... 8

4 Requirements ................................................................................................................................................... 8

5 Test methods .................................................................................................................................................... 9

5.1 Principle ............................................................................................................................................................. 9

5.2 Materials and reagents ............................................................................................................................... 10

5.2.1 Test organisms .............................................................................................................................................. 10

5.2.2 Culture media and reagents ..................................................................................................................... 10

5.2.3 Test surface .................................................................................................................................................... 13

5.3 Apparatus and glassware .......................................................................................................................... 14

5.4 Preparation of test organism suspensions and product test solutions .................................... 15

5.4.1 Test organism suspensions ...................................................................................................................... 15

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

5.5 Procedure........................................................................................................................................................ 19

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

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

5.5.3 Counting of the test mixtures ................................................................................................................... 24

5.6 Calculation and expression of results ................................................................................................... 25

5.6.1 Elaboration of data: counting of weighed average values ............................................................. 25

5.6.2 Verification of methodology ..................................................................................................................... 26

5.6.3 Expression of results ................................................................................................................................... 27

5.6.4 Conclusion....................................................................................................................................................... 27

5.7 Test report ...................................................................................................................................................... 28

Annex A (informative) Corresponding referenced strains .......................................................................... 30

Annex B (informative) Neutralizers ..................................................................................................................... 32

Annex C (informative) Expression of results with the dilution-neutralization method.................... 34

Annex D (informative) Bactericidal activity in surfaces in general use conditions (for clean

conditions) ...................................................................................................................................................... 36

Annex E (informative) Precision of the test result .......................................................................................... 38

Bibliography ................................................................................................................................................................. 39

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SIST EN 13697:2015+A1:2019
EN 13697:2015+A1:2019 (E)
European foreword

This document (EN 13697:2015+A1:2019) 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 January 2020, and conflicting national standards shall

be withdrawn at the latest by January 2020.

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.

This document includes Amendment 1 approved by CEN on 10 June 2019.
This document supersedes !EN 13697:2015".

The start and finish of text introduced or altered by amendment is indicated in the text by tags ! ".

!The changes between EN 13697:2015+A1:2019 and EN 13697:2001 are the following:

— interfering substance has been changed from 0,03 % bovine albumin to 0,85 % skimmed milk (see

Clause 4, Table 1) for Pseudomonas aeruginosa under clean conditions only;

— A. brasiliensis (ex A. niger) spore preparation has been updated in order to harmonize this step with

the QST fungicidal test method amendment issued in 2012 (see 5.4.1.3 b));

— calculations of the weighed means and of the results have been modified in order to be harmonized

with new CEN TC 216 standards (see 5.4.1.5, 5.5.2, 5.5.3 and 5.6);

— other paragraphs have been harmonized to new CEN TC 216 standards (e.g. preparation of hard

water, 5.2.2.7);
— deletion of obligatory and additional conditions (see Table 1 and 5.5.1);
— update of Bovine albumin and skimmed solutions preparations (see 5.2.2.8.2);
— add of instruction for using vacuum desiccator;

— update on bacteria working culture preparation (see 5.4.1.2) and counting of bacterial and fungal

test suspensions (see 5.4.1.4);

— clarification to the determination of microbicidal concentrations by updating 5.5.2.1 b) and adding

pictures of carriers;

— updates on dilution preparation for fungal and bacterial strains (see 5.5.2.2, 5.5.2.3, 5.5.2.4) and

counting of test mixtures (5.5.3).
Data from EN 13697:2015 are still valid with the exception of:
— Pseudomonas aeruginosa and Candida albicans under clean conditions."
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SIST EN 13697:2015+A1:2019
EN 13697:2015+A1:2019 (E)

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, Republic of North Macedonia, France,

Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands,

Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and

the United Kingdom.
---------------------- Page: 6 ----------------------
SIST EN 13697:2015+A1:2019
EN 13697:2015+A1:2019 (E)
Introduction

This European Standard describes a surface test method for establishing whether a product proposed

as a disinfectant in the fields described in Clause 1 has or does not have bactericidal and/or fungicidal

or yeasticidal activity on non-porous surfaces.

This European Standard has been revised in order to modify the interfering substance under “clean

conditions” adopted for P. aeruginosa; in order to modify the calculation of N, NC, NT, Nc, Na and

consequently the final results and to harmonize the standard with the other recent CEN TC 216

standards.

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

time, temperature, organisms on surfaces ...) reflect parameters which are found in practical situations

including conditions which may influence the action of disinfectants. Each use concentration found from

this test corresponds to defined experimental conditions.

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

product types.

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

additional test conditions need to be used.
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SIST EN 13697:2015+A1:2019
EN 13697:2015+A1:2019 (E)
1 Scope

This European Standard specifies a test method (phase 2/step 2) and the minimum requirements for

bactericidal and/or fungicidal or yeasticidal activity of chemical disinfectants that form a homogeneous

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

food, industrial, domestic and institutional areas, excluding areas and situations where disinfection is

medically indicated and excluding products used on living tissues.
The scope of this European Standard applies at least to the following:
a) Processing, distribution and retailing of:
1) Food of animal origin:
i) milk and milk products;
ii) meat and meat products;
iii) fish, seafood and products;
iv) eggs and egg products;
v) animal feeds;
vi) etc.
2) Food of vegetable origin:
i) beverages;
ii) fruits, vegetables and derivatives (including sugar distillery);
iii) flour, milling and backing;
iv) animal feeds;
v) etc.
b) Institutional and domestic areas:
1) catering establishments;
2) public areas;
3) public transports;
4) schools;
5) nurseries;
6) shops;
7) sports rooms;
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SIST EN 13697:2015+A1:2019
EN 13697:2015+A1:2019 (E)
8) waste container (bins);
9) hotels;
10) dwellings;
11) clinically non sensitive areas of hospitals;
12) offices;
13) etc.
c) Other industrial areas:
1) packaging material;
2) biotechnology (yeast, proteins, enzymes...);
3) pharmaceutical;
4) cosmetics and toiletries;
5) textiles;
6) space industry, computer industry;
7) etc.

Using this European Standard, it is possible to determine the bactericidal or fungicidal or yeasticidal

activity of the undiluted product. As three concentrations are tested, in the active to non active range,

dilution of the product is required and, therefore, the product forms a homogeneous stable preparation

in hard water.

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

recommendations.

NOTE 1 The method described is intended to determine the activity of commercial formulations or active

substances on bacteria and/or fungi in the conditions in which they are used.

NOTE 2 This method cannot be used to evaluate the activity of products against mycobacteria.

2 Normative references

The following documents, in whole or in part, are normatively referenced in this document and are

indispensable for its application. For dated references, only the edition cited applies. For undated

references, the latest edition of the referenced document (including any amendments) applies.

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

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

(including bacteriophages) activity

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

disinfectants and antiseptics

ISO 4793, Laboratory sintered (fritted) filters — Porosity grading, classification and designation

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SIST EN 13697:2015+A1:2019
EN 13697:2015+A1:2019 (E)
3 Terms and definitions

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

4 Requirements

The product shall demonstrate at least a 4 decimal log (lg) reduction for bacteria and at least a 3

decimal log (lg) reduction for fungi, when tested in accordance with Table 1 and 5.5.1.

Table 1 — Experimental conditions
Yeasticidal activity on Fungicidal activity on
Bactericidal activity on
non-porous surfaces non-porous surfaces
Test conditions non-porous surfaces
without mechanical without mechanical
without mechanical action
action action
Test organisms
Enterococcus hirae
(see 5.2.1)
Escherichia coli Candida albicans
minimum
Candida albicans
spectrum of Pseudomonas aeruginosa Aspergillus brasiliensis
test
Staphylococcus aureus
organisms
Saccharomyces cerevisiae
Salmonella typhimurium
(for breweries)
Test organisms
additional Lactobacillus brevis any relevant test organism
Saccharomyces cerevisiae
(examples)
var. diastaticus (for
Enterobacter cloacae
breweries)
In a range from (4 ± 1) °C In a range from (4 ± 1) °C
In a range from (4 ± 1) °C to
to (40 ± 1) °C to (40 ± 1) °C
(40 ± 1) °C
For tests performed at For tests performed at
Test temperature
For tests performed at room
room temperature, the room temperature, the
temperature, the range shall
range shall be between range shall be between
be between 18 °C and 25 °C
18 °C and 25 °C 18 °C and 25 °C
in a range from 1 min to 60 in a range from 1 min to in a range from 1 min to 60
min (from 1 min to 5 min at 60 min (from 1 min to 5 min (from 1 min to 5 min

Contact time intervals of 1 min and from min at intervals of 1 min at intervals of 1 min and

5 min to 60 min at intervals and from 5 min to 60 min from 5 min to 60 min at
of 5 min) at intervals of 5 min) intervals of 5 min)
0,3 g/l bovine albumin for
Interfering
Staphylococcus aureus, 0,3 g/l bovine albumin for
0,3 g/l bovine albumin for
substance
Enterococcus hirae, C. albicans and A.
C. albicans
Escherichia coli and brasiliensis
clean conditions
Pseudomonas aeruginosa
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EN 13697:2015+A1:2019 (E)
Yeasticidal activity on Fungicidal activity on
Bactericidal activity on
non-porous surfaces non-porous surfaces
Test conditions non-porous surfaces
without mechanical without mechanical
without mechanical action
action action
3,0 g/l bovine albumin for
Interfering
Staphylococcus aureus, 3,0 g/l bovine albumin for
3,0 g/l bovine albumin for
substance
Enterococcus hirae, C. albicans and A.
C. albicans
Pseudomonas aeruginosa brasiliensis
dirty conditions
and Escherichia coli
Interfering
substance
any relevant substance any relevant substance any relevant substance
additional
Log reduction
from a water
≥ 4 lg ≥ 3 lg ≥ 3 lg
control (decimal
lg)

The referenced test conditions (General purposes) are by no means intended as requirements for the use

of a product, nor as requirements for the evaluation and acceptance of products by regulatory authorities.

The application time for the product is specified by the manufacturer.

If specific applications have to be considered, the bactericidal/yeasticidal/fungicidal activity has to be

determined additionally under relevant conditions concerning application time, temperature, strains and

Interfering Substances.
5 Test methods
5.1 Principle

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

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

which covers the dried film. The surface is maintained at a specified temperature for a defined period of

time. The surface is transferred to a previously validated neutralization medium so that the action of

the disinfectant is immediately neutralized. The number of surviving organisms which can be recovered

from the surface is determined quantitatively.

The number of bacteria or fungi on a surface treated with hard water in place of the disinfectant is also

determined and the reduction in viable counts attributed to the product is calculated by difference.

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5.2 Materials and reagents
5.2.1 Test organisms
The bactericidal activity shall be evaluated using the following four strains:
— Pseudomonas aeruginosa 1
ATCC 15 442 ;
— Staphylococcus aureus ATCC 6 538;
— Enterococcus hirae ATCC 10 541;
— Escherichia coli ATCC 10 536.

The fungicidal or yeasticidal activity shall be evaluated using the following two strains:

— Candida albicans ATCC 10 231;
— Aspergillus brasiliensis (ex A. niger) ATCC 16 404.

If required for specific applications, additional strains may be chosen from, for example:

— Salmonella typhimurium ATCC 13 311;
— Lactobacillus brevis DSM 6 235;
— Enterobacter cloacae DSM 6 234;
— Saccharomyces cerevisiae (for breweries) or ATCC 9 763 or DSM 1 333;
— Saccharomyces cerevisiae var. diastaticus (for DSM 70 487.
breweries)

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

If additional strains are used, they shall be incubated under optimum growth conditions (temperature,

time, atmosphere) and noted in the test report.

If the additional strains selected do not correspond to the specified strains, their suitability for

supplying inocula of sufficient concentration shall be verified. If the additional strains tested 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 under a reference for 5 years.

5.2.2 Culture media and reagents
5.2.2.1 General

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

5.2.2.2 Water

The water shall be free from substances that are toxic or inhibiting to bacteria and fungi. It shall be

freshly glass distilled and not demineralized water.

ATCC 15 442, ATCC 6 538, ATCC 10 541, ATCC 10 536, ATCC 10 231, ATCC 16 404 and ATCC 13311 are the collection

numbers of strains supplied by the American Type Culture Collections. This information is given for the convenience of users of

this standard and does not constitute an endorsement by CEN of the product named. Equivalent products can be used if they

can be shown to lead to the same results.
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Sterilize in the autoclave (see 5.3.2.1).

NOTE 1 If the water is sterilized during sterilization of the reagents, this is not necessary.

NOTE 2 If distilled water of adequate quality is not available, water for injectable preparation (see European

Pharmacopoeia) can be used.
5.2.2.3 Tryptone Soya Agar (TSA)
For maintenance of bacterial strains and performance of viable counts.
Tryptone, pancreatic digest of casein 15,0 g
Soya peptone, papaic digest of Soybean meal 5,0 g
NaCl 5,0 g
Agar 15,0 g
Water (see 5.2.2.2) 1 000,0 ml

Sterilize in the autoclave (see 5.3.2.1). After sterilization, the pH of the medium shall be equivalent to

7,2 ± 0,2 when measured at 20 ° C.
5.2.2.4 Malt extract agar (MEA)
For maintenance of fungal strains, sporulation and performance of viable counts.
Malt extract (food grade, e.g. Cristomalt powder from 30,0 g
Difal)
Agar 15,0 g
Water (see 5.2.2.2) 1 000,0 ml

The malt extract should be food grade (e.g. Cristomalt powder from Difal) or equivalent that is not

highly purified and not only based on maltose (e.g. Malt extract from OXOID) . However, if there are

problems producing at least 75 % spiny spores see 5.4.1.4.2.

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

5,6 ± 0,2 when measured at (20 ± 1) °C.

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

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

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

CEN of the product named. Equivalent products may be used if they can be shown to lead to the same results.

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5.2.2.5 Diluent
Tryptone sodium chloride solution:
Tryptone, pancreatic digest of casein 1,0 g
NaCl 8,5 g
Water (see 5.2.2.2) 1 000,0 ml

Sterilize in the autoclave (see 5.3.2.1). After sterilization the pH shall be equivalent to 7,0 ± 0,2 when

measured at 20 °C.
5.2.2.6 Neutralizer

The neutralizer shall be validated for the product under test in accordance with 5.5.2.3 and 5.5.2.4. The

neutralizer shall be sterile.

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

given in Annex B.
5.2.2.7 Hard water for dilution of the products
Hard water for dilution of products shall be prepared as follows:

— solution A: Dissolve 19,84 g anhydrous MgCl and 46,24 g anhydrous CaCl in water (see 5.2.2.2)

2 2

and dilute to 1 000 ml. Sterilize by membrane filtration (5.3.2.19) or in the autoclave (5.3.2.1 a).

Autoclaving – if used - may cause a loss of liquid. In this case, make up to 1 000 ml with water

(5.2.2.2) under aseptic conditions. Store the solution in the refrigerator (5.3.2.15) for no longer

than one month;

— solution B: Dissolve 35,02 g NaHC0 in water (see 5.2.2.2) and dilute to 1 000 ml. !deleted text"

Sterilize by membrane filtration (5.3.2.19). Store the solution in the refrigerator (5.3.2.15) for no

longer than one week.

Add at least 600 ml water (see 5.2.2.2) to 6,0 ml of solution A in a 1 000 ml volumetric flask, then add

8,0 ml solution B. Mix and dilute to 1 000 ml with water (see 5.2.2.2).

Sterilize by passing through a filter with a maximum effective pore size of 0,45 µm.

The pH of the hard water shall be 7,0 ± 0,2, when measured at (20 ± 1) °C (5.3.2.6). If necessary, adjust

the pH by using a solution of approximately 40 g/l (about 1 mol/l) of sodium hydroxide (NaOH) or

approximately 36,5 g/l (about 1 mol/l) of hydrochloric acid (HCl).

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

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

produces a different final water hardness expressed as calcium carbonate (CaCO3) in each test tube. In any case,

the final hardness is lower than 375 mg/l of calcium carbonate.
5.2.2.8 Interfering substances
5.2.2.8.1 General

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

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

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

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

detergents) shall be fully defined.

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

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

report (see 5.7).
5.2.2.8.2 !Bovine albumin solutions"
!Bovine albumin solutions for the test conditions shall be prepared as follows:
— Preparation for clean conditions:

— dissolve 0,06 g of bovine albumin (Cohn fraction V for Dubos medium) in 100 ml of water (see

5.2.2.2);

— sterilize by 0,45 µm membrane filtration (see 5.3.2.19), keep in the refrigerator and use within

one month.

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

— Preparation for dirty conditions:

— dissolve 0,6 g of bovine albumin (Cohn fraction V for Dubos medium) in 100 ml of water (see

5.2.2.2);

— sterilize by 0,45 µm membrane filtration (see 5.3.2.19), keep in the refrigerator and u

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