SIST EN 13697:2002
(Main)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/step2)
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/step2)
This European Standard specifies a test method (phase 2/step 2) and the minimum requirements for bactericidal and/or fungicidal activity of chemical disinfectants that form a homogeneous physically stable preparation in hard water and that are used 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 :
- milk and milk products ;
- meat and meat products ;
- fish, seafood and products ;
- eggs and egg products ;
- animal feeds ;
- etc.
2) Food of vegetable origin :
- beverages ;
- fruits, vegetables and derivatives (including sugar distillery) ;
- flour, milling and backing ;
- animal feeds ;
- etc.
b) Institutional and domestic areas :
- catering establishments ;
- public areas ;
- public transports ;
- schools ;
- nurseries ;
- shops ;
- sports rooms ;
- waste container (bins) ;
- hotels ;
- dwellings ;
- clinically non sensitive areas of hospitals ;
- offices ;
- etc.
c) Other industrial areas :
- packaging material ;
- biotechnology (yeast, proteins, enzymes...) ;
- pharmaceutical ;
- cosmetics and toiletries ;
- textiles ;
- space industry, computer industry ;
- etc.
Using this European Standard, it is possible to determine the bactericidal or fungicidal activity of the undiluted product. As three concentrations have to be tested, in the active to non active range, dilution of the product is required and, therefore, the product shall form a homogeneous stable preparation in hard water.
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.
Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflächen-Versuch zur Bestimmung der bakteriziden und/oder fungiziden Wirkung chemischer Desinfektionsmittel in den Bereichen Lebensmittel, Industrie, Haushalt und öffentliche Einrichtungen - Prüfverfahren ohne mechanische Behandlung und Anforderungen (Phase 2/Stufe 2)
Diese Europäische Norm legt ein Prüfverfahren (Phase 2/Stufe 2) und Anforderungen an die bakterizide und/oder fungizide Mindestwirkung von chemischen Desinfektionsmitteln fest, die in hartem Wasser eine homogene, physikalisch stabile Lösung ergeben, und die im Lebensmittelbereich, in der Industrie, im Haushalt und in öffentlichen Einrichtungen benutzt werden, ausgenommen davon sind Bereiche und Situationen, in denen die Desinfektion medizinisch indiziert ist, und Produkte, die auf lebenden Geweben benutzt werden.
Diese Europäische Norm gilt grundsätzlich für den folgenden Anwendungsbereich:
a) Verarbeitung, Vertrieb und Verkauf von:
1) Lebensmitteln tierischer Herkunft:
- Milch und Milchprodukte;
- Fleisch und Fleischprodukte;
- Fisch, Meerestiere und daraus hergestellte Zubereitungen;
- Eier und Eiprodukte;
- Tiernahrung;
- usw.
2) Lebensmitteln pflanzlicher Herkunft:
- Getränke;
- Früchte, Gemüse und daraus hergestellte Zubereitungen (einschließlich Zucker, Destillationsprodukte);
- Mehl, gemahlene und gebackene Produkte;
- Tiernahrung;
- usw.
b) Öffentliche Einrichtungen und Haushaltsbereiche:
- Verpflegungseinrichtungen;
- Öffentliche Bereiche;
- Öffentliche Verkehrsmittel;
- Schulen;
- Kindergärten;
- Geschäfte;
- Sportstätten;
- Abfallbehälter (Mülltonnen);
- Hotels;
- Wohngebäude;
- Klinisch nicht relevante Bereiche von Krankenhäusern;
- Büroräume;
- usw.;
c) Andere industrielle Bereiche:
- Verpackungsmaterial;
- Biotechnologie (Hefe, Proteine, Enzyme usw.);
- Hersteller von Pharmazeutika;
- Hersteller von Kosmetika und Toilettenartikeln;
- Hersteller von Textilien;
- Weltraumforschungsindustrie und Computerindustrie;
- usw.
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 domestiques et en collectivité - Méthode d'essai sans action mécanique et prescriptions (phase 2/étape 2)
La présente Norme européenne décrit une méthode d'essai (phase 2, étape 2) et les prescriptions minimales relatives a l'activité bactéricide et/ou fongicide des désinfectants chimiques qui forment une préparation homogene, physiquement stable dans l'eau dure et qui sont utilisés dans le domaine de l'agro-alimentaire, dans l'industrie, dans le domaine domestique et en collectivités, a l'exception des domaines et des situations ou la désinfection est médicalement prescrite et a 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) fabrication, distribution et vente au détail de :
1) produits alimentaires d'origine animale :
- lait et produits laitiers ;
- viande et produits dérivés de la viande ;
- poisson, fruits de mer et leurs produits dérivés ;
- ?ufs et produits avicoles ;
- produits alimentaires pour animaux ;
- etc. ;
2) produits alimentaires d'origine végétale :
- boissons ;
- fruits, légumes et leurs dérivés (y compris le sucre, les produits de distillerie ?) ;
- farine, minoterie et boulangerie ;
- produits alimentaires pour animaux ;
- etc. ;
b) domaines domestiques et collectivité :
- établissements de restauration;
- lieux publics ;
- transports publics ;
- écoles ;
- haltes-garderies ;
- magasins ;
- salles de sport ;
- conteneurs pour déchets (poubelles ?) ;
- hôtels ;
- lieux d'habitation ;
- zones cliniquement non sensibles des hôpitaux;
- bureaux ;
- etc. ;
c) autres secteurs industriels :
- matériaux d'emballage ;
- biotechnologie (levure, protéines, enzymes, ?.). ;
- pharmaceutique ;
- cosmétique et produits d'hygiene corporelle ;
- textiles ;
- industrie spatiale, industrie informatique ;
- etc..
Kemična razkužila in antiseptiki - Kvantitativni preskus na neporoznih površinah za ocenjevanje 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)
General Information
Relations
Standards Content (Sample)
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflächen-Versuch zur Bestimmung der bakteriziden und/oder fungiziden Wirkung chemischer Desinfektionsmittel in den Bereichen Lebensmittel, Industrie, Haushalt und öffentliche Einrichtungen - Prüfverfahren ohne mechanische Behandlung und Anforderungen (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 domestiques et en collectivité - Méthode d'essai sans action mécanique et prescriptions (phase 2/étape 2)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/step2)71.100.35Kemikalije za dezinfekcijo v industriji in domaChemicals for industrial and domestic disinfection purposesICS:Ta slovenski standard je istoveten z:EN 13697:2001SIST EN 13697:2002en01-september-2002SIST EN 13697:2002SLOVENSKI
STANDARD
SIST EN 13697:2002
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 13697August 2001ICS 11.080.20; 71.100.35English versionChemical disinfectants and antiseptics - Quantitative non-poroussurface test for the evaluation of bactericidal and/or fungicidalactivity of chemical disinfectants used in food, industrial,domestic and institutional areas - Test method and requirementswithout mechanical action (phase 2/step2)Antiseptiques et désinfectants chimiques - Essai quantitatifde surface non-poreuse pour l'évaluation de l'activitébactéricide et/ou fongicide des désinfectants chimiquesutilisés dans le domaine de l'agro-alimentaire, dansl'industrie, dans les domaines domestiques et encollectivité - Méthode d'essai sans action mécanique etprescriptions (phase 2/étape 2)Chemische Desinfektionsmittel und Antiseptika -Quantitativer Oberflächen-Versuch zur Bestimmung derbakteriziden und/oder fungiziden Wirkung chemischerDesinfektionsmittel in den Bereichen Lebensmittel,Industrie, Haushalt und öffentliche Einrichtungen -Prüfverfahren ohne mechanische Behandlung undAnforderungen (Phase 2/Stufe 2)This European Standard was approved by CEN on 25 July 2001.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the 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 translationunder the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2001 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 13697:2001 ESIST EN 13697:2002
EN 13697:2001 (E)2ContentspageIntroduction.41Scope.52Normative references.63Terms and definitions.64Requirements.74.1Requirements for bactericidal activity on surfaces.74.2Requirements for fungicidal activity on surfaces.74.3Requirements for bactericidal and fungicidal activity on surfaces.85Test method.85.1Principle.85.2Materials and reagents.85.3Apparatus and glassware.125.4Preparation of bacterial and fungal suspensions and test solutions.135.5Procedure.165.6Calculation and expression of results.185.7Test report.19Annex A (normative)
Test for validation of neutralisation.21A.1Principle.21A.2Preparation of neutralisation test suspensions and test surfaces.21A.3Preparation of test solution.21A.4Test for validation of neutralisation.21A.5Counting procedure.21A.6Validation.22Annex B (informative)
Neutralisers.23Annex C (informative)
Example of a typical test report.24Annex D (informative)
Corresponding referenced strains.26Annex E (informative)
Information on the application and interpretation of European Standards onchemical disinfectants and antiseptics.27E.1General guidelines for the application and interpretation of test methods in accordance withEuropean Standards for chemical disinfectants and antiseptics.27E.2Guide to interpretation of tests for chemical disinfectants and antiseptics.28Bibliography.29SIST EN 13697:2002
EN 13697:2001 (E)3ForewordThis European Standard has been prepared by Technical Committee CEN/TC 216, "Chemical disinfectants andantiseptics", 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 orby endorsement, at the latest by February 2002, and conflicting national standards shall be withdrawn at the latestby February 2002.Annexes designated "normative" are part of the body of the standard. Annexes designated "informative" are givenonly for information.Annex A is normative.Annex B, C, D and E are informative.According to the CEN/CENELEC Internal Regulations, the national standards organizations of the followingcountries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland,France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden,Switzerland and the United Kingdom.SIST EN 13697:2002
EN 13697:2001 (E)4IntroductionThis European Standard describes a surface test method for establishing whether a product proposed as adisinfectant in the fields described in clause 1 has or does not have bactericidal and/or fungicidal activity onnon-porous surfaces.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 includingconditions which may influence the action of disinfectants. Each use concentration found from this test correspondsto defined experimental conditions.The conditions are intended to cover general purposes and to allow reference between laboratories and producttypes.However for some applications the recommendations of use of a product can differ and therefore additional testconditions need to be used.SIST EN 13697:2002
EN 13697:2001 (E)51 ScopeThis European Standard specifies a test method (phase 2/step 2) and the minimum requirements for bactericidaland/or fungicidal activity of chemical disinfectants that form a homogeneous physically stable preparation in hardwater and that are used in food, industrial, domestic and institutional areas, excluding areas and situations wheredisinfection 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 : milk and milk products ; meat and meat products ; fish, seafood and products ; eggs and egg products ; animal feeds ; etc.2) Food of vegetable origin : beverages ; fruits, vegetables and derivatives (including sugar distillery) ; flour, milling and backing ; animal feeds ; etc.b) Institutional and domestic areas : catering establishments ; public areas ; public transports ; schools ; nurseries ; shops ; sports rooms ; waste container (bins) ; hotels ; dwellings ; clinically non sensitive areas of hospitals ;SIST EN 13697:2002
EN 13697:2001 (E)6 offices ; etc.c) Other industrial areas : packaging material ; biotechnology (yeast, proteins, enzymes.) ; pharmaceutical ; cosmetics and toiletries ; textiles ; space industry, computer industry ; etc.Using this European Standard, it is possible to determine the bactericidal or fungicidal activity of the undilutedproduct. As three concentrations have to be tested, in the active to non active range, dilution of the product isrequired and, therefore, the product shall form a homogeneous stable preparation in hard water.NOTE 1The method described is intended to determine the activity of commercial formulations or active substances onbacteria and/or fungi in the conditions in which they are used.NOTE 2This method corresponds to a phase 2/step 2 test (see annex E).2 Normative referencesThis European Standard incorporates by dated or undated reference, provisions from other publications. Thesenormative references are cited at the appropriate places in the text, and the publications are listed hereafter. Fordated references, subsequent amendments to or revisions of any of these publications apply to this EuropeanStandard only when incorporated in it by amendment or revision. For undated references the latest edition of thepublication referred to applies (including amendments).EN 1040, Chemical disinfectants and antiseptics - Basic bactericidal activity - Test method and requirements(phase 1).EN 1275, Chemical disinfectants and antiseptics - Basic fungicidal activity - Test method and requirements(phase 1).EN 10088-1, Stainless steels – Part 1: List of stainless steel.EN 10088-2, Stainless steels. Technical delivery conditions for sheet/plate and strip for general purposes.EN 12353, Chemical disinfectants and antiseptics - Preservation of microbial strains used for the determination ofbactericidal and fungicidal activity.3 Terms and definitionsFor the purposes of this European Standard, the following terms and definitions apply :3.1product (for chemical disinfection and/or antisepsis)chemical agent or formulation used as a chemical disinfectant [EN 1040]3.2bactericideproduct which kills vegetative bacteria under defined conditions [EN 1040]SIST EN 13697:2002
EN 13697:2001 (E)7NOTEThe adjective derived from "bactericide" is "bactericidal".3.3bactericidal activity on surfacescapability of a product to produce at least a 104 reduction in the number of viable bacterial cells belonging toreference strains of Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Enterococcus hiraewithin 5 min at 20 °C under conditions defined by this European Standard3.4fungicideproduct which kills fungi including their spores under defined conditions [EN 1275]3.5fungicidal activity on surfacescapability of a product to produce at least a 103 reduction in number of viable fungi belonging to reference strainsof Candida albicans and Aspergillus niger within 15 min at 20 °C under conditions defined by this EuropeanStandard3.6clean conditionsconditions representative of surfaces which have received a satisfactory cleaning programme and/or are known tocontain minimal levels of organic and/or inorganic materials [EN 1276]3.7dirty conditionsconditions representative or surfaces which are known to or may contain, organic and/or inorganic materials[EN 1276]4 Requirements4.1 Requirements for bactericidal activity on surfacesThe product, when diluted in hard water and tested in accordance with clause 5, under simulated clean conditions(0,3 g/l bovine albumin and 1g/l tryptone, see 3.6) or dirty conditions (3 g/l bovine albumin and 1g/l tryptone,see 3.7) according to its practical applications and under the required test conditions shall demonstrate at least a104 reduction in viable bacterial countsFor a claim of bactericidal activity on surfaces for general purposes, the bactericidal activity shall be evaluatedusing the following four strains : Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus hirae andEscherichia coli at a temperature between 18°C ± 1°C and 25°C ± 1°C and a contact time of 5 min.The determined bactericidal concentration of the test product is suggested as being suitable for practical situationsof use.Where appropriate (specific purposes), additional specific bactericidal activity shall be determined under otherconditions of time, temperature, additional strains and interfering substances (see 5.2.1 and 5.2.2.8) in accordancewith 5.5.1 in order to take into account intended specific use conditions.NOTEFor these additional conditions the concentration defined as a result can be lower than the one obtained underthe initial conditions, at a temperature between 18 °C ± 1 °C and 25 °C ± 1 °C, 5 min, 4 selected reference strains.Products under test shall at least possess a bactericidal activity as specified in EN 1040.4.2 Requirements for fungicidal activity on surfacesThe product, when diluted in hard water and tested in accordance with clause 5, under simulated clean conditions(0,3 g/l bovine albumin and 1g/l tryptone, see 3.6) or dirty conditions (3 g/l bovine albumin, see 3.7) according to itspractical applications and under the required test conditions shall demonstrate at least a 103 reduction in viablefungal counts.SIST EN 13697:2002
EN 13697:2001 (E)8For a claim of fungicidal activity on surfaces for general purposes, the fungicidal activity shall be evaluated usingthe following two strains : Candida albicans, and Aspergillus niger at a temperature between 18 °C ± 1 °C and25 °C ± 1 °C and a contact time of 15 min.Where appropriate (specific purposes), additional specific fungicidal activity shall be determined under otherconditions of time, temperature, additional strains and interfering substances (see 5.2.1 and 5.2.2.8) in accordancewith 5.5.1 in order to take into account intended specific use conditions.NOTEFor these additional conditions the concentration defined as a result can be lower than the one obtained under theinitial conditions at a temperature between 18 °C ± 1 °C and 25 °C ± 1 °C, 15 min, 2 selected reference strains.Products under test shall at least possess a fungicidal activity as specified in EN 1275.4.3 Requirements for bactericidal and fungicidal activity on surfacesThe product, when diluted in hard water and tested in accordance with clause 5, under simulated clean conditions(0,3 g/l bovine albumin and 1g/l tryptone, see 3.6) or dirty conditions (3 g/l bovine albumin, see 3.7) according to itspractical applications and under the required test conditions shall demonstrate at least a 104 reduction in viablebacterial counts (Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus hirae, Escherichia coli) and a103 reduction in viable fungal counts (Candida albicans and Aspergillus niger).The determined bactericidal and/or fungicidal concentration of the tested product is suggested as being suitable forgeneral, practical situations of use (see 5.6.3).Where appropriate, additional specific bactericidal and/or fungicidal activity shall be determined under otherconditions of time, temperature, additional strains and interfering substances (see 5.2.1 and 5.5.1) in accordancewith 5.5.1 in order to take into account intended specific use conditions.NOTEFor these additional conditions the concentration defined as a result can be lower than the one obtained under theinitial conditions, at a temperature between 18 °C ± 1 °C and 25 °C ± 1 °C, 5 min and/or 15 min, with selected reference strains.This European Standard is applicable only to disinfectants and antiseptics, complying with EN 1040 (for bactericidalactivity) and/or EN 1275 (for fungicidal activity).5 Test method5.1 PrincipleA test suspension of bacteria or fungi in a solution of interfering substances is inoculated onto a test stainless steelsurface 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 apreviously validated neutralisation medium so that the action of the disinfectant is immediately neutralised. Thenumber 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 determinedand the reduction in viable counts attributed to the product is calculated by difference.5.2 Materials and reagents5.2.1 Test organismsThe bactericidal activity shall be evaluated using the following four strains : Pseudomonas aeruginosaATCC 15 442 1) ;
1)ATCC 15 442, ATCC 6 538, ATCC 10 541, ATCC 10 536, ATCC 10 231, ATCC 16 404 and ATCC 13311 are the collectionnumbers of strains supplied by the American Type Culture Collections. DSM 6 235, DSM 6 234, DSM 1333 and DSM 70487 arethe collection numbers of strains supplied by the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen). Thisinformation is given for the convenience of users of this standard and does not constitute an endorsement by CEN of theproduct named. Equivalent products can be used if they can be shown to lead to the same results.SIST EN 13697:2002
EN 13697:2001 (E)9 Staphylococcus aureusATCC 6 538 ; Enterococcus hiraeATCC 10 541 ; Escherichia coliATCC 10 536.The fungicidal activity shall be evaluated using the following two strains : Candida albicansATCC 10 231 ; Aspergillus nigerATCC 16 404.If required for specific applications, additional strains may be chosen from ; for example : Salmonella typhimuriumATCC 13 311 ; Lactobacillus brevisDSM 6 235 ; Enterobacter cloacaeDSM 6 234 ; Saccharomyces cerevisiae (for breweries) orATCC 9 763 or DSM 1 333 ; Saccharomyces cerevisiae var. diastaticus (for breweries)DSM 70 487.NOTESee annex D 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 ofsufficient concentration shall be verified. If the additional strains tested are not classified at a reference centre, theiridentification characteristics shall be stated. In addition, they shall be held by the testing laboratory or nationalculture under a reference for 5 years.5.2.2 Culture media and reagents5.2.2.1 GeneralThe reagents shall be of analytical grade and/or appropriate for microbiological purposes.NOTETo improve the reproducibility of the results, it is recommended that commercially available dehydrated materialshould be used whenever possible for the preparation of culture media. The manufacturer's instructions relating to thepreparation of these products should be rigorously followed.5.2.2.2 WaterThe water shall be free from substances that are toxic or inhibiting to bacteria and fungi. It shall be freshly glassdistilled and not demineralized water.Sterilise in the autoclave (see 5.3.2.1).NOTE 1If the water is sterilized during sterilisation of the reagents, this is not necessary.NOTE 2If distilled water of adequate quality is not available, water for injectable preparation (see European Pharmacopoeia)can be used.SIST EN 13697:2002
EN 13697:2001 (E)105.2.2.3 Tryptone Soya Agar (TSA)For maintenance of bacterial strains and performance of viable counts.Tryptone, pancreatic digest of casein15,0 gSoya peptone, papaic digest of Soybean meal5,0 gNaCl5,0 gAgar15,0 gWater (see 5.2.2.2)1 000,0 mlSterilise in the autoclave (see 5.3.2.1). After sterilisation the pH of the medium shall be equivalent to 7,2 ± 0,2 whenmeasured at 20
C.5.2.2.4 Malt extract agar (MEA)For maintenance of fungal strains, sporulation and performance of viable counts.Malt extract (technical grade)30,0 gSoya peptone3,0 gAgar15,0 gWater (see 5.2.2.2)1 000,0 mlSterilise in the autoclave (see 5.3.2.1). After sterilisation the pH of the medium shall be equivalent to 5,6 ± 0,2 whenmeasured at 20 °C.5.2.2.5 DiluentTryptone sodium chloride solution :Tryptone, pancreatic digest of casein1,0 gNaCl8,5 gWater (see 5.2.2.2)1 000,0 mlSterilise in the autoclave (see 5.3.2.1). After sterilisation the pH shall be equivalent to 7,0 ± 0,2 when measured at20 °C.5.2.2.6 NeutraliserThe neutraliser shall be validated for the product under test in accordance with annex A. The neutraliser shall besterile.NOTEInformation on neutralisers 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 productsHard water for dilution of products shall be prepared as follows : solution A : Dissolve 19,84 g anhydrous MgCl2 and 46,24 g anhydrous CaCl2 in water (see 5.2.2.2) and diluteto 1 000 ml ;SIST EN 13697:2002
EN 13697:2001 (E)11 solution B : Dissolve 35,02 g NaHC03 in water (see 5.2.2.2) and dilute to 1000 ml.Add at least 600 ml water (see 5.2.2.2) to 6,0 ml of solution A in a 1000 ml volumetric flask, then add 8,0 mlsolution B and dilute to 1 000 ml with water (see 5.2.2.2).Sterilise by passing through a filter with a maximum effective pore size of 0,45 µm.After adjustment, the pH of the solution shall be 7,0 ± 0,2 before use.The solution shall be stored at 4 °C to 8 °C for a maximum holding time of one month.NOTEWhen preparing the three concentrations of product test solutions (see 5.4.3) the addition of the product in thishard water solution produces a different final water hardness in each test tube. In any case the final hardness is lower than300 mg/kg of CaC03 in the test tube.5.2.2.8 Interfering substances5.2.2.8.1 GeneralThe ionic composition (pH, calcium and/or magnesium hardness) and chemical composition (mineral substances,protein, carbohydrates, lipids, detergents etc) shall be fully defined.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.The method of preparation and sterilisation together with the composition shall be noted in the test report (see 5.7).5.2.2.8.2 Bovine albumin solutionsBovine albumin solutions for the test conditions shall be prepared as follows :a) preparation for clean conditions ; dissolve 0,060 g of bovine albumin (Cohn fraction V for Dubos medium) in 100 ml of water (see 5.2.2.2) ; sterilise by membrane filtration ;The final concentration of bovine albumin in the test procedure (see 5.5.2) is 0,3 g/l.b) preparation for dirty conditions ; dissolve 0,60 g of bovine albumin (Cohn fraction V for Dubos medium) in 100 ml of water (see 5.2.2.2) ; sterilize by membrane filtration ;The final concentration of bovine albumin in the test procedure (see 5.5.2) is 3 g/l.In addition, other interfering substances can be chosen from :5.2.2.8.3 Milk (dairies.)Skimmed milk, guaranteed free of antibiotics or additives, shall be reconstituted at a rate of 100 g powder per litreof water (see 5.2.2.2). The working solution shall be prepared as follows : prepare a solution of 2,0 % (V/V) in water (see 5.2.2.2) by adding 2 parts of reconstituted milk to 98 parts ofwater ; sterilize for 30 min at 105 °C ± 3 °C (or 5 min at 121 °C ± 3 °C).The final concentration of milk in the test procedure (see 5.5.2) shall be 1,0 % (V/V) of reconstituted milk.SIST EN 13697:2002
EN 13697:2001 (E)125.2.2.8.4 Yeast extract (breweries.)Dehydrated yeast extract for bacteriology, shall be prepared as follows : prepare a 20 g/l solution in water (see 5.2.2.2), adjust to pH 7,0 ± 0,2 with sodium hydroxide ; sterilize in the autoclave (see 5.3.1).The final concentration of yeast extract in the test procedure (see 5.5.2) shall be 10 g/l.5.2.2.8.5 Sucrose (beverage, soft drink industries)Prepare a 20 g/l solution in water (see 5.2.2.2), sterilize by membrane filtration.The final concentration of sucrose in the test procedure (see 5.5.2) is 10 g/l.5.2.2.8.6 pH 5,0 and pH 9,0 buffer solutions (clean-in-place.)The buffer solution shall be described in the test report and pH values shall be recorded. The final pH in the testshall be equal to 5,0 ± 0,2 or 9,0 ± 0,2.5.2.2.8.7 Sodium lauryl sulphate (cosmetics.)Prepare a 10 g/l solution of sodium lauryl sulphate in water (see 5.2.2.2). Sterilize in the autoclave (see 5.3.1).The final concentration of sodium lauryl sulphate in the test procedure (see 5.5.2) is 5 g/l.5.2.3 Test surfaceThese shall be 1.4301 (EN 10088-1) stainless steel discs (2 cm diameter discs) with Grade 2 B in accordance withthe requirements of EN 10 088-2 finish on both sides 2). The surfaces shall be as flat as possible and this is bestachieved by using stainless steel of a gauge of 1,2 mm or 1,5 mm. The surfaces shall be used once only andsubsequently discarded.Prior to use, place the surfaces should be placed in a beaker (minimum size : 50 ml) containing not less than 20 mlof 5 % (V/V)
Decon 3) for 60 min. Immediately rinse the discs with running freshly distilled water for 10 s.The surface shall not be allowed to dry to any extent. The discs shall only be handled with forceps. Rinse the discswith water (see 5.2.2.2) for a further 10 s to ensure complete removal of the surfactant. To supply a satisfactoryflow of water, a sterilized fluid dispensing pressure vessel with suitable hose and connectors or other suitablemethod can be used and regulated to supply approximately 2 000 ml per min. To sterilise place the clean disc in abath containing 70 % (V/V) iso-propanol for 15 min. Remove the disc and dry by evaporation under laminar air flow.5.3 Apparatus and glassware5.3.1 GeneralSterilize all glassware and parts of apparatus that will come into contact with the culture media and reagents or thesample, except those which are supplied sterile, by one of the following methods :a) in the autoclave (see 5.3.2.1) by maintaining it at 121
C0 3 for a minimum holding time of 15 min ;b) in the dry heat sterilizer (see 5.3.2.1) by maintaining it at 180 °C for a minimum holding time of 30 min, at170 °C for a minimum holding time of 1 h or at 160 °C for a minimum holding time of 2 h.
2)Suitable stainless steel discs can usually be purchased from local engineering companies.3)Decon is an example of a suitable product available commercially. This information is given for the convenience of usersof this standard and does not constitute an endorsement by CEN of this product.SIST EN 13697:2002
EN 13697:2001 (E)135.3.2 Usual microbiological laboratory equipment 4) and in particular, the followin
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.