1、November 2014 Translation by DIN-Sprachendienst.English price group 9No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).IC
2、S 87.040!%)E“2250634www.din.deDDIN EN 15458Paints and varnishes Laboratory method for testing the efficacy of film preservatives in acoating against algae;English version EN 15458:2014,English translation of DIN EN 15458:2014-11Beschichtungsstoffe Laborverfahren fr die Prfung der Wirksamkeit von Fil
3、mkonservierungsmitteln in einerBeschichtung gegen Algen;Englische Fassung EN 15458:2014,Englische bersetzung von DIN EN 15458:2014-11Peintures et vernis Mthode dessai en laboratoire permettant de dterminer lefficacit des prservateurs dufeuil dun revtement contre les algues;Version anglaise EN 15458:
4、2014,Traduction anglaise de DIN EN 15458:2014-11SupersedesDIN EN 15458:2007-10www.beuth.deDocument comprises 14 pagesIn case of doubt, the German-language original shall be considered authoritative.10.14 DIN EN 15458:2014-11 2 A comma is used as the decimal marker. National foreword This document (E
5、N 15458:2014) has been prepared by Technical Committee CEN/TC 139 “Paints and varnishes” (Secretariat: DIN, Germany). The responsible German body involved in its preparation was the DIN-Normenausschuss Beschichtungsstoffe und Beschichtungen (DIN Standards Committee Coatings and Coating Materials), W
6、orking Committee NA 002-00-15 AA Bautenbeschichtungen. Amendments This standard differs from DIN EN 15458:2007-10 as follows: a) a new term “active substance” has been introduced (3.1); b) the designations of the algal cultures has been updated; c) the standard has been editorially revised. Previous
7、 editions DIN EN 15458: 2007-10 EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 15458 August 2014 ICS 87.040 Supersedes EN 15458:2007English Version Paints and varnishes - Laboratory method for testing the efficacy of film preservatives in a coating against algae Peintures et vernis - Mthode de
8、ssai en laboratoire permettant de dterminer lefficacit des prservateurs du feuil dun revtement contre les algues Beschichtungsstoffe - Laborverfahren fr die Prfung der Wirksamkeit von Filmkonservierungsmitteln in einer Beschichtung gegen Algen This European Standard was approved by CEN on 10 July 20
9、14. 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
10、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-CEN
11、ELEC 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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Ita
12、ly, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Ave
13、nue 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 15458:2014 E2 Contents Page Foreword 3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Principle 5 5 Apparatus and
14、materials .5 6 Microorganisms .6 7 Sampling and preparation of test samples and of specimens .6 7.1 Sampling .6 7.2 Preparation of test samples (see Annex A) 6 7.3 Conditioning of the test samples .7 7.4 Preparation and number of specimens .7 8 Procedure .7 8.1 Preparation of Bolds Basal Medium 7 8.
15、2 Preparation of the stock solutions .7 8.3 Preparation of the Petri dishes with culture medium 8 8.4 Preparation of stock cultures and sub-cultures .8 8.5 Preparation of the algal suspension 8 8.6 Inoculation and incubation (see Annex A) 9 8.7 Assessment 9 9 Test report 9 Annex A (informative) Labo
16、ratory method for testing the efficacy of film preservatives in a coating against algae . 11 Bibliography . 12 DIN EN 15458:2014-11 EN 15458:2014 (E) 3 Foreword This document (EN 15458:2014) has been prepared by Technical Committee CEN/TC 139 “Paints and varnishes”, the secretariat of which is held
17、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 th
18、at 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 15458:2007. According to the CEN-CENELEC Internal Regulations, the national standards organization
19、s 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, Malta,
20、Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. DIN EN 15458:2014-11 EN 15458:2014 (E) 4 Introduction This document identifies criteria to assess the efficacy of film preservatives in a coating against algae. The results
21、of the 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
22、of the 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. DIN EN 15458:2014-11 EN 15458:2014 (E) 5 1 Scope This European Standard specifies
23、a laboratory test method for determining the biocidal/biostatic efficacy of single active substances or combinations thereof used in film preservatives in a coating against algal growth. The standard does not apply to coatings not susceptible to algal growth. The test method comprises only active su
24、bstances 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 ar
25、e not in 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 ind
26、ispensable 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 23270, Paints and varn
27、ishes 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 Terms and definitions For the purposes of this document, the following term and definition applies. 3.1 act
28、ive 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 article “a“ between “or“ and “micro-organism“ was deleted 4 Principle For the determination of the algicidal
29、 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 algal suspension and incubated over a certain period of time under conditions appropriate for algal growth. Conclus
30、ions can be drawn with regard to the algicidal efficacy of the film preservatives in a coating from the intensity of the algal growth on the coated surface of the specimen after incubation. The method described in this standard is a semiquantitative, comparative method between coatings with and with
31、out film preservatives. 5 Apparatus and materials 5.1 Cutting device for preparing the specimens (coated filter paper, with a diameter of 55 mm). 5.2 Autoclave for sterilization. 5.3 Incubator, capable of maintaining (23 2) C. 5.4 Pipette, in the range between 100 l to 1 000 l, with sterile tips or
32、combi-tips of 12,5 ml. 5.5 Filter paper without biocidal effect (e.g. cellulose with a pore size of 0,45 m and a thickness of 650 m). DIN EN 15458:2014-11 EN 15458:2014 (E) 6 5.6 Automatic welding apparatus to seal the bags. 5.7 Sterilized glass bottles (100 ml, 0,5 l, 1 l). 5.8 Sterilized test tube
33、s or other sterilized glassware for preparing the slant agar cultures. 5.9 Bold modified Basal medium as specified in the method (see 8.1). 5.10 Stock solution (see 8.2). 5.11 Culture flask with cap (0,5 l or 1 l). 5.12 Laboratory balance, capable of weighing to an accuracy of 0,1 g. 5.13 Microscope
34、 5.14 Device to determine cell count (commercially available counting chamber, e.g. Thoma chamber). 5.15 Device for applying the coating 5.16 Sterile Petri dishes (with a diameter of 94 mm and a height of 16 mm). 5.17 Sterile tweezers 5.18 Sterile water 5.19 Class 1 microbiological safety cabinet ac
35、cording to EN 12469. 5.20 Luxmeter 5.21 Cold white or daylight lamp 6 Microorganisms Blue-green algae Nostoc commune SAG1)1453-3; Blue-green algae Gloeocapsa atrata Ktzing (syn. Anacystis montana) CCAP2)1430/1; Green algae Klebsormidium flaccidum SAG 335-5; Green algae Stichococcus bacillaris SAG 37
36、9-1a = CCAP 379/1A. From these four microorganisms one blue-green and one green algae are selected. 7 Sampling and preparation of test samples and of specimens 7.1 Sampling Take a representative sample of the coating material or coating system for testing in accordance with EN ISO 1513. 7.2 Preparat
37、ion of test samples (see Annex A) 1)SAG = (Sammlung von Algenkulturen (Culture Collection of Algae), Gttingen; available at: Georg-August Universitt Gttingen, Germany. 2)CCAP = Culture Collection of Algae and Protozoa; SAMS Research Services Ltd, Scottish Marine institute Oban, Scotland, UK. DIN EN
38、15458:2014-11 EN 15458:2014 (E) 7 Coat a strip of filter paper without biocidal effect with the coating material/system to be tested. The application rate shall be that recommended by the coating manufacturer for normal use. 7.3 Conditioning of the test samples Condition the test samples in a horizo
39、ntal 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 the coating material and end use corresponding to information given by the manufacturer. 7.4 Preparation and number of specimens After conditi
40、oning, three specimens, each of a diameter of 55 mm shall be prepared from the test samples. The specimens shall be sealed in a plastics bag and sterilized using gamma radiation of 10 kGy. Other methods of sterilization may be agreed upon between the parties. For each test series, three specimens co
41、ated with coating material containing the film preservative, three specimens coated with the same coating material without film preservative and three specimens of the uncoated substrate shall be tested. 8 Procedure 8.1 Preparation of Bolds Basal Medium 3)For the algal nutritive solution the followi
42、ng substances are required: a) 10 ml each of stock solutions a) to f) in 8.2; b) 1 ml each of trace element stock solutions g) to j) in 8.2; c) 940 ml demineralized or distilled water; d) 15 g agar (only for the solid nutritive medium). The solution shall be sterilized in the autoclave. For the test
43、 both solid (with 1,5 % agar) and also liquid nutritive medium are required. 8.2 Preparation of the stock solutions Stock solutions: a) NaNO310,0 g Distilled water 400 ml b) CaCl22H2O 1,0 g Distilled water 400 ml c) MgSO47H2O 3,0 g Distilled water 400 ml d) K2HPO43,0 g Distilled water 400 ml e) KH2P
44、O47,0 g Distilled water 400 ml f) NaCl 1,0 g Distilled water 400 ml Trace element stock solutions: g) Ethylenediaminetetraacetic acid 50 g KOH 31 g Distilled water 1 000 ml h) FeSO47H2O 4,98 g Distilled water (acidified) 1 000 ml 3)Bischoff, H. W. Gloeocapsa atrata Ktzing (syn. Anacystis montana) CC
45、AP 1430/1, Klebsormidium flaccidum SAG 335-5, Stichococcus bacillaris SAG 379-1a = CCAP 379/1A) sub-cultures shall be prepared on agar slants for use as stock cultures. The incubation takes place at room temperature and under illumination, using a cycle of 16 h illumination and 8 h darkness. Fluores
46、cent tubes of the type daylight or white light shall be used, at a distance of about 50 cm at about (1 000 200) lx. Alternatively, Bolds Basal Medium (according to 8.1) without agar can be used to prepare liquid cultures. NOTE 1 From experience it is known that the stock cultures will have grown in
47、about 2 weeks to such an extent that sub-cultures can be prepared from them. Sufficient stock cultures should always be kept in reserve. Sub-cultures: From the stock cultures sub-cultures shall be prepared in conical flasks containing liquid nutritive medium, followed by incubation as described abov
48、e. NOTE 2 Experience shows that these sub-cultures will have grown after 7 days to 14 days, to such an extent that they can be used further. Cultures with filamentous algae can be shaken up somewhat more frequently, to loosen and disintegrate the filaments. 8.5 Preparation of the algal suspension Before starting the tests, 200 ml of an actively-growing, ready-to-use individual algal culture shall be mixed with 200 ml of sterile algal nutrient solution. These two components shall be mixed thorough
