1、BSI Standards PublicationBS EN 15457:2014Paints and varnishes Laboratory method fortesting the efficacy of filmpreservatives in a coatingagainst fungiBS EN 15457:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 15457:2014. Itsupersedes BS EN 15457:2007 which
2、 is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee STI/28, Paint systems for non-metallic substrates.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryp
3、rovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2014. Published by BSI StandardsLimited 2014ISBN 978 0 580 84804 9ICS 87.040Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published
4、 under the authority of theStandards Policy and Strategy Committee on 31 August 2014.Amendments issued since publicationDate Text affectedBS EN 15457:2014EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 15457 August 2014 ICS 87.040 Supersedes EN 15457:2007English Version Paints and varnishes - L
5、aboratory method for testing the efficacy of film preservatives in a coating against fungi Peintures et vernis - Mthode dessai en laboratoire permettant de dterminer lefficacit des prservateurs du feuil dun revtement contre les champignons Beschichtungsstoffe - Laborverfahren fr die Prfung der Wirks
6、amkeit von Filmkonservierungsmitteln in einer Beschichtung gegen Pilze This European Standard was approved by CEN on 10 July 2014. 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 stand
7、ard 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 an
8、y 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 Repu
9、blic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN
10、 COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 15457:2014 EBS EN 15
11、457:2014EN 15457:2014 (E) 2 Contents Page Foreword 3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Principle 5 5 Apparatus and materials .6 6 Fungi .6 6.1 Fungi more likely to grow in an exterior environment .6 6.2 Fungi more likely to grow in an interior environmen
12、t7 7 Sampling and preparation of test samples and of specimens .7 7.1 Sampling .7 7.2 Preparation of test samples (see Annex A) 7 7.3 Conditioning of the test samples .7 7.4 Preparation and number of specimens .7 8 Procedure .8 8.1 Preparation of the Petri dishes with the culture medium 8 8.2 Prepar
13、ation of stock cultures and sub-cultures .8 8.3 Preparation of the spore suspension 8 8.4 Inoculation and incubation (see Annex A) 8 8.5 Assessment 8 9 Test report 9 Annex A (informative) Laboratory method for testing the efficacy of film preservatives in a coating against fungi 10 Annex B (informat
14、ive) Designation of the percentage area of disfigurements according to EN 16492:2014, Table A.3 . 11 Bibliography . 12 BS EN 15457:2014EN 15457:2014 (E) 3 Foreword This document (EN 15457:2014) has been prepared by Technical Committee CEN/TC 139 “Paints and varnishes”, the secretariat of which is he
15、ld by DIN. 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 February 2015 and conflicting national standards shall be withdrawn at the latest by February 2015. Attention is drawn to the possibility
16、 that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. This document supersedes EN 15457:2007. According to the CEN-CENELEC Internal Regulations, the national standards organizat
17、ions of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malt
18、a, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. BS EN 15457:2014EN 15457:2014 (E) 4 Introduction This document identifies criteria to assess efficacy of film preservatives in a coating against fungi. The results of the
19、 method allow evaluation of an active substance with regard to its inclusion in Annex I of the Biocidal Products Directive 98/8/EC (Directive 98/8/EC of the European Parliament and of the Council of 16 February 1998 concerning the placing of biocidal products on the market BPD) or in the list of the
20、 Biocidal Product Regulation (BPR, Regulation (EU) 528/2012). The characteristics of the biocide treated coating material should conform to national regulations with regard to health, safety and the environment. BS EN 15457:2014EN 15457:2014 (E) 5 1 Scope This European Standard specifies a laborator
21、y test method for determining the biocidal/biostatic efficacy of single active substances or combinations thereof used in film preservatives in a coating against fungal growth. This standard does not apply to coatings not susceptible to fungal growth. The test method comprises only active substances
22、 for film preservation, not the protection of the substrate itself, e.g. wood, which is dealt with in another standard. The test method is applicable for active substances used for wood and masonry coatings. It is not applicable to marine coatings. Safety, health and environmental aspects are not in
23、 the scope of this standard. Determination of the performance of film preservatives in coatings by applying ageing procedures is not within the scope of this standard. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensab
24、le 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 12469, Biotechnology - Performance criteria for microbiological safety cabinets EN 16492:2014, Paints and varnish
25、es - Evaluation of the surface disfigurement caused by fungi and algae on coatings EN 23270, Paints and varnishes and their raw materials - Temperatures and humidities for conditioning and testing (ISO 3270) EN ISO 1513, Paints and varnishes - Examination and preparation of test samples (ISO 1513) 3
26、 Terms and definitions For the purposes of this document, the following term and definition applies. 3.1 active substance substance or micro-organism that has an action on or against harmful organisms SOURCE: Biocidal Product Regulation (BPR, Regulation (EU) 528/2012), Article 3.1 (c), modified the
27、article “a“ between “or“ and “micro-organism“ was deleted 4 Principle To determine the fungicidal efficacy of film preservatives in a coating, the coating material is applied to a substrate conditioned according to EN 23270, placed onto an agar surface, inoculated with a standard fungal spore suspen
28、sion and incubated. Conclusions can be drawn to the fungicidal efficacy of the film preservatives in a coating from the intensity of the fungal growth on the surface of the specimen after incubation. The method described here is a semiquantitative, comparative method between coatings, with and witho
29、ut film preservatives. BS EN 15457:2014EN 15457:2014 (E) 6 5 Apparatus and materials 5.1 Cutting device for preparing the specimens (coated filter paper with a diameter of 55 mm). 5.2 Autoclave 5.3 Incubator capable of maintaining (24 2) C. 5.4 Pipette, in the range between 100 l to 1 000 l, with st
30、erile tips or combi-tips of 12,5 ml. 5.5 Filter paper without fungicidal effect (e.g. cellulose with a pore size of 0,45 m and a thickness of 650 m). 5.6 Automatic welding apparatus to seal the bags. 5.7 Sterilized glass bottles (100 ml). 5.8 Laboratory balance, capable of weighing to an accuracy of
31、 0,1 g. 5.9 Microscope 5.10 Device to determine cell count (commercially available counting chamber, e.g. Thoma chamber). 5.11 Wetting agent (e.g. N-Methyltaurine). 5.12 Device for applying the coating 5.13 Sterilized test tubes or other sterilized glassware for preparing slant agar cultures. 5.14 S
32、terile Drigalski spatula 5.15 Sterile platinum loop 5.16 Sterile glass funnel with cotton wool 5.17 Sterile Petri dishes (with a diameter of 94 mm, and a height of 16 mm). 5.18 Sterile tweezers 5.19 Sterile water 5.20 Class 2 microbiological safety cabinet according to EN 12469. 6 Fungi 6.1 Fungi mo
33、re likely to grow in an exterior environment a) Aureobasidium pullulans (DSM1)2404) b) Alternaria alternata (DSM 62010) 1)DSM = DSMZ = Deutsche Sammlung fr Mikroorganismen und Zellkulturen (German collection of micro organisms and cell cultures), Braunschweig, Germany. BS EN 15457:2014EN 15457:2014
34、(E) 7 c) Cladosporium cladosporioides (DSM 62121) d) Phoma violaceae (IMI2)49948ii) e) Ulocladium atrum (IMI 79906 or DSMZ 63068) 6.2 Fungi more likely to grow in an interior environment a) Aspergillus versicolor (DSM 1943) b) Aspergillus niger (DSM 12634) c) Stachybotrys chartarum (DSMZ 2144) d) Pe
35、nicillium purpurogenum (DSM 62866) e) Rhodotorula mucilaginosa (DSM 70825) The spore suspension used for the test shall be a mixture containing two fungi selected from the first group (6.1) and two fungi selected from the second group (6.2). 7 Sampling and preparation of test samples and of specimen
36、s 7.1 Sampling Take a representative sample of the coating material or of the coating system for testing in accordance with EN ISO 1513. 7.2 Preparation of test samples (see Annex A) Coat a strip of filter paper without biocidal effect with the coating material to be tested. The application rate sha
37、ll be that recommended by the coating manufacturer for normal use. 7.3 Conditioning of the test samples Condition the test sample in a horizontal position for at least 5 days at (23 2) C and (50 5) % relative humidity, in accordance with EN 23270. NOTE The conditioning time might vary according to t
38、he coating material and end use corresponding to information given by the manufacturer. 7.4 Preparation and number of specimens After conditioning, three specimens each with a diameter of 55 mm shall be prepared from the test samples. The specimens shall be sealed in a plastics bag and sterilized us
39、ing gamma radiation of 10 kGy. Other methods of sterilization may be agreed between the parties. For each test series three specimens coated with coating material containing the film preservative, three specimens coated with the same coating material without film preservative and three specimens of
40、the uncoated substrate shall be tested. 2)IMI = CABI = Bioscience Genetic Resource Collection, Egham, UK. BS EN 15457:2014EN 15457:2014 (E) 8 8 Procedure 8.1 Preparation of the Petri dishes with the culture medium A malt (3 %)-agar(1,5 %)-medium shall be sterilized in the autoclave. After cooling th
41、e medium to 55 C to 60 C, 20 ml shall be poured into each sterile Petri dish under aseptic conditions. 8.2 Preparation of stock cultures and sub-cultures Sub-cultures shall be obtained by inoculating spore material from a stock culture to freshly prepared agar slope culture media and shall be used f
42、or preparing the spore suspension. From these sub-cultures further sub-cultures can be derived in sufficient number. After the inoculation the sub-cultures shall be incubated at (24 2) C until good sporulation has been achieved. This might require 3 days to 7 days, depending on the fungal species us
43、ed for testing. The sub-cultures can be stored satisfactorily at 3 C to 7 C for a period of 3 months. 8.3 Preparation of the spore suspension For preparing the spore suspension a well-sporulating sub-culture is used. To this sub-culture 5 ml of sterile deionized water should be added (if required al
44、so add a surfactant e.g. 0,1 % N-Methyltaurine). The spores shall be carefully washed down from the agar slope, using a platinum loop as an aid, filtered through a sterile glass funnel with cotton wool and collected in a sterile glass bottle. After counting the spore concentration by using a commerc
45、ially available counting chamber and diluting each with sterile water to 106to 107spores/ml all spore suspensions intended for testing shall be mixed in equal parts. 8.4 Inoculation and incubation (see Annex A) In addition to the coated and uncoated specimens (see 7.4) a further three Petri dishes c
46、ontaining the nutritive agar medium only shall be inoculated. The sterilized specimens shall be placed centrally onto the surface of the culture media using sterile tweezers. The coated surface of the specimen shall be face up and there shall be full contact without air bubbles between the specimens
47、 and the surface of the culture medium. Should the coating lead to an undulation of the filter paper the paper should be kept even and in close contact with the agar by appropriate means. Otherwise a different substrate may be used. Check that the substrate does not inhibit growth of each selected t
48、est organism under the test conditions. If the substrate does inhibit growth it cannot be used. Under aseptic conditions in a safety cabinet the specimens shall be uniformly inoculated with 0,2 ml each of the mixed spore suspension using a suitable pipette. It is permissible to use up to an addition
49、al 0,8 ml of diluent to ensure an even distribution over the surface. Afterwards the suspension is spread out onto the agar plate with Drigalski spatula. The surface of the specimen shall remain free of agar medium during this operation. Following inoculation incubate the agar plates at (24 2) C. 8.5 Assessment Assess the fungal growth 7 days, 14 days and 21 days after the inoculation, using the assessment scale EN 16492:2014, Table A.3 (see also Annex B).The assessment is carried out visually macroscopically. NOTE 1 If required a microsco
