EN 13624:2003

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Chemische Desinfektionsmittel und Antiseptika - Quantitativer Suspensionsversuch zur Prüfung der fungiziden Wirkung chemischer Desinfektionsmittel für Instrumente im humanmedizinischen Bereich - Prüfverfahren und Andforderungen (Phase 2, Stufe 1)Antiseptiques et désinfectants chimiques - Essai quantitatif de suspension pour l'évaluation de l'activité fongicide des désinfectants chimiques utilisés pour les instruments en médecine - Méthode d'essai et exigences (phase 2, étape 1)Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of fungicidal activity of chemical disinfectants for instruments used in the medical area - Test method and requirements (phase 2, step 1)11.080.20Dezinfektanti in antiseptikiDisinfectants and antisepticsICS:Ta slovenski standard je istoveten z:EN 13624:2003SIST EN 13624:2004en,fr,de01-februar-2004SIST EN 13624:2004SLOVENSKI
STANDARD
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 13624December 2003ICS 11.080.20English versionChemical disinfectants and antiseptics - Quantitative suspensiontest for the evaluation of fungicidal activity of chemicaldisinfectants for instruments used in the medical area - Testmethod and requirements (phase 2, step 1)Antiseptiques et désinfectants chimiques - Essai quantitatifde suspension pour l'évaluation de l'activité fongicidedesdésinfectants chimiques utilisés pour les instruments enmédecine - Méthode d'essai et prescriptions (phase 2,étape 1)Chemische Desinfektionsmittel und Antiseptika -Quantitativer Suspensionsversuch zur Prüfung derfungiziden Wirkung chemischer Desinfektionsmittel fürInstrumente im humanmedizinischen Bereich -Prüfverfahren und Andforderungen (Phase 2, Stufe 1)This European Standard was approved by CEN on 3 November 2003.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,Hungary, Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Slovakia, Spain, Sweden, Switzerland and UnitedKingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2003 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 13624:2003 ESIST EN 13624:2004

Referenced strains in national collections.27Annex B (informative)
Neutralizer and Rinsing liquids.28B.1Neutralizers.28B.2Rinsing liquids.29B.3Neutralizer added to the agar for counting :.29Annex C (informative)
Flow diagrams.30C.1Dilution neutralization method.30C.2Membrane filtration method.31Annex D (informative)
Example of a typical test report.34Annex E (informative)
Information on the application and interpretation of European standards onchemical disinfectants and antiseptics.36E.1Introduction.36E.2General guidelines for the application and interpretation of test methods in accordancewith European Standards for chemical disinfectants and antiseptics.36E.3Guide to the interpretation of tests for chemical disinfectants and antiseptics.37Annex ZA (informative)
Clauses of this European Standard addressing essential requirements orother provisions of EU Directives.38Bibliography.39SIST EN 13624:2004

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 standard and does not constitute an endorsement by CEN ofthe product named.SIST EN 13624:2004

2)Disposable equipment is an acceptable alternative to reusable glassware.3)Vortex® in an example of a suitable product available commercially. This information is given for the convenience ofusers of this standard and does not constitute an endorsement by CEN of this product.SIST EN 13624:2004

4)cfu/ml = colony forming unit(s) per millilitreSIST EN 13624:2004

5)Malassez is an example of a suitable product available commercially. This information is given for the convenience ofusers of this standard and does not constitute an endorsement by CEN of this product.SIST EN 13624:2004

and 30 min ;¾ allowed deviation for each chosen contact time is ± 10 s.c) Interfering substance :¾ obligatory interfering substance to be tested is 0,30 g/l bovine albumin (see 5.2.2.8.2) under clean ora mixture of 3 ml/l sheep erythrocytes and 3,0 g/l bovine albumin (see 5.2.2.8.3) under dirtyconditions – according to practical applications ;¾ additional interfering substances may be tested according to specific field of application.
d) Test organisms :¾ obligatory test organisms are: Candida albicans and Aspergillus niger or only Candida albicans (see5.2.1).5.5.1.2 Choice of test methodThe method of choice is the dilution-neutralization method (see 5.5.2). To determine a suitable neutralizercarry out the validation of the dilution neutralization method (see 5.5.2.3, 5.5.2.4 and 5.5.2.5 in connectionwith 5.5.2.6) using a neutralizer, chosen according to laboratory experience and published data.If this neutralizer is not valid, repeat the validation test using an alternative neutralizer containing acombination of polysorbate 80
(30 g/l), saponin (30 g/l), L-histidine (1 g/l), lecithin (3 g/l), sodium thiosulfate(5 g/l) in either diluent (see 5.2.2.4) or in phosphate buffer 0,0025 mol/l (see B.1).If both neutralizers are found to be invalid, the membrane filtration method (see 5.5.3) may be used in place ofthe dilution-neutralization method.NOTEIn special circumstances it may be necessary to add neutralizer to MEA (see 5.2.2.3).If neutralizer is added to MEA the same amount shall be added to MEA used in the test procedure.5.5.1.3 Validation and control procedures - General instructionsThe neutralization and/or removal of the fungicidal and/or fungistatic activity of the product shall be controlledand validated - only for the highest product test concentration - for each of the used test organisms and foreach experimental condition (interfering substance, temperature, contact time). These procedures(experimental condition control, neutralizer or filtration control and method validation) shall be performed atthe same time with the test and with the same neutralizer – or rinsing liquid – used in the test.In the case of ready-to-use-products use water (see 5.2.2.2) instead of hard water.If because of problems with neutralization a neutralizer has been added to MEA (see 5.5.1.3) used for thevalidation and control procedures the MEA used for the test shall contain the same amount of this neutralizeras well.5.5.1.4 Equilibration of temperaturePrior to testing, equilibrate all reagents (product test solutions (see 5.4.2), test suspension (see 5.4.1.4),validation suspension (see 5.4.1.5), diluent (see 5.2.2.4), hard water (see 5.2.2.7) interfering substances(see 5.2.2.8) to the test temperature of q
using the water bath (see 5.3.2.2) controlled at q. Check that thetemperature of the reagents is stabilized at q.The neutralizer (see 5.2.2.5) or the rinsing liquid (see 5.2.2.6) and water (see 5.2.2.2) shall be equilibrated ata temperature of 20 °C ± 1 °C.SIST EN 13624:2004

(see 5.5.1.1 a) for 2 min ± 10 s. At the end of this time, add 8,0 ml of one ofthe product test solutions (see 5.4.2). Restart the stopwatch at the beginning of the addition, mix (see5.3.2.6) and place the tube in a water bath controlled at q for the chosen contact time t (see 5.5.1.1 b).Just before the end of t, mix (see 5.3.2.6) again ;b) At the end of t take a 1,0 ml sample of the test mixture Na and transfer into a tube containing 8,0 mlneutralizer (see 5.2.2.5) and 1,0 ml water (see 5.2.2.2). Mix (see 5.3.2.6) and place in a water bathcontrolled at 20 °C . After a neutralization time of 5 min ± 10 s, immediately take a sample of 1,0 ml of theneutralized test mixture Na (containing neutralizer, product test solution, interfering substance, testsuspension) in duplicate and inoculate using the pour plate or the spread plate technique ;c) Perform the procedure a) to b) using the other product test solutions at the same time ;d) Perform the procedure a) to c) applying the other obligatory and - if appropriate - other additionalexperimental conditions (see 5.5.1.1.).5.5.2.3 Experimental condition control “A” (Validation of the selected experimental conditions orverification of the absence of any lethal effect in the test conditions)a) Pipette 1,0 ml of the interfering substance used in the test (see 5.5.2.2) into a tube. Add 1,0 ml of thevalidation suspension (see 5.4.1.5). Start the stopwatch immediately, mix (see 5.3.2.6) and place the tube min ± 10 s. At the end of this time add 8,0 ml of hard water(see 5.2.2.7). (In the case of ready-to-use products: water (see 5.2.2.2) instead of hard water). Restartthe stopwatch at the beginning of the addition, mix (see 5.3.2.6) and place the tube in a water batht. Just before the end of t, mix (see 5.3.2.6) again;b) At the end of t, take a sample of 1,0 ml of this mixture A in duplicate and inoculate using the pour plate orthe spread plate technique (see 5.5.2.2b).Incubation and counting see 5.5.2.6.
6)For a graphical representation of this method see C.1SIST EN 13624:2004
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