1、BSI Standards PublicationBS EN 15458:2014Paints and varnishes Laboratory method fortesting the efficacy of filmpreservatives in a coatingagainst algaeBS EN 15458:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 15458:2014. Itsupersedes BS EN 15458:2007 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2、 is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee STI/28, Paint systems for non-metallic substrates.A list of organizations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 84803 2ICS 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 15458:2014EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 15458 August 2014 ICS 87.040 Supersedes EN 15458:2007English Version Paints and varnishes - L
5、aboratory method for testing the efficacy of film preservatives in a coating against algae Peintures et vernis - Mthode dessai en laboratoire permettant de dterminer lefficacit des prservateurs du feuil dun revtement contre les algues Beschichtungsstoffe - Laborverfahren fr die Prfung der Wirksamkei
6、t von Filmkonservierungsmitteln in einer Beschichtung gegen Algen 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 standard w
7、ithout 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 oth
8、er 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,
9、 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 COMM
10、ITTEE 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 15458:2014 EBS EN 15458:2
11、014EN 15458:2014 (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 .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
12、(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 8.2 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-cult
13、ures .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) Laboratory method for testing the efficacy of film preservatives in a coating against algae . 11 Bibliography . 12 BS EN 15458:2014EN 15458:2014 (E)
14、 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 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
15、 latest by February 2015 and conflicting national standards shall be withdrawn at the latest by February 2015. Attention is drawn to the possibility 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
16、 all such patent rights. This document supersedes EN 15458:2007. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Est
17、onia, 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 the United Kingdom. BS EN 15458:2014EN 154
18、58:2014 (E) 4 Introduction This document identifies criteria to assess the efficacy of film preservatives in a coating against algae. The results 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/E
19、C 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 Biocidal Product Regulation (BPR, Regulation (EU) 528/2012). The characteristics of the biocide treated coating material should conform to national
20、 regulations with regard to health, safety and the environment. BS EN 15458:2014EN 15458:2014 (E) 5 1 Scope This European Standard specifies a laboratory test method for determining the biocidal/biostatic efficacy of single active substances or combinations thereof used in film preservatives in a co
21、ating against algal growth. The standard does not apply to coatings not susceptible to algal growth. The test method comprises only active substances 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 f
22、or active substances used for wood and masonry coatings. It is not applicable to marine coatings. Safety, health and environmental aspects are 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 o
23、f this standard. 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 (in
24、cluding any amendments) applies. EN 12469, Biotechnology - Performance criteria for microbiological safety cabinets 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 prepara
25、tion of test samples (ISO 1513) 3 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
26、), Article 3.1 (c), modified the article “a“ between “or“ and “micro-organism“ was deleted 4 Principle For the determination of the algicidal 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, i
27、noculated with a standard algal suspension and incubated over a certain period of time under conditions appropriate for algal growth. Conclusions 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
28、the specimen after incubation. The method described in this standard is a semiquantitative, comparative method between coatings with and without film preservatives. 5 Apparatus and materials5.1 Cutting device for preparing the specimens (coated filter paper, with a diameter of 55 mm). 5.2 Autoclave
29、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 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). BS EN 15458:2014EN 15458:2
30、014 (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 tubes 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
31、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 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 (w
32、ith 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 according 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
33、Ktzing (syn. Anacystis montana) CCAP2)1430/1; Green algae Klebsormidium flaccidum SAG 335-5; Green algae Stichococcus bacillaris SAG 379-1a = 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 S
34、ampling Take a representative sample of the coating material or coating system for testing in accordance with EN ISO 1513. 7.2 Preparation of test samples (see Annex A) 1)SAG = (Sammlung von Algenkulturen (Culture Collection of Algae), Gttingen; available at: Georg-August Universitt Gttingen, German
35、y. 2)CCAP = Culture Collection of Algae and Protozoa; SAMS Research Services Ltd, Scottish Marine institute Oban, Scotland, UK. BS EN 15458:2014EN 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 r
36、ecommended by the coating manufacturer for normal use. 7.3 Conditioning of the test samples Condition the test samples 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 the coating
37、material and end use corresponding to information given by the manufacturer. 7.4 Preparation and number of specimens After conditioning, 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 ra
38、diation of 10 kGy. Other methods of sterilization may be agreed upon 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 the un
39、coated substrate shall be tested. 8 Procedure 8.1 Preparation of Bolds Basal Medium 3)For the algal nutritive solution the following 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 di
40、stilled water; d) 15 g agar (only for the solid nutritive medium). The solution shall be sterilized in the autoclave. For the test 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
41、 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) KH2PO47,0 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
42、ml h) FeSO47H2O 4,98 g Distilled water (acidified) 1 000 ml 3)Bischoff, H. W. Gloeocapsa atrata Ktzing (syn. Anacystis montana) CCAP 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. T
43、he incubation takes place at room temperature and under illumination, using a cycle of 16 h illumination and 8 h darkness. Fluorescent 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) w
44、ithout agar can be used to prepare liquid cultures. NOTE 1 From experience it is known that the stock cultures will have grown in 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 cult
45、ures sub-cultures shall be prepared in conical flasks containing liquid nutritive medium, followed by incubation as described above. 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 alg
46、ae 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 com
47、ponents shall be mixed thoroughly, but without introducing contamination, so as to disperse cells and break-up filaments. The resulting suspension should be slightly coloured and contain approximately 106cfu/ml4). To obtain a mixture of different algal species for test use, combine the required indi
48、vidual algal cultures with sterile algal nutrient solution in the above proportions (e.g. two different test species will require 2 200 ml algal suspension plus 2 200 ml sterile algal nutrient solution = 800 ml total). 4)cfu = colony forming units. BS EN 15458:2014EN 15458:2014 (E) 9 8.6 Inoculation
49、 and incubation (see Annex A) In addition to the coated and uncoated specimens (see 7.4) a further three Petri dishes containing the nutritive medium only shall be inoculated. In a safety cabinet the sterilized specimens shall be placed centrally onto the solid algal nutritive medium in the Petri dishes. The coated surface of the specimen shall be face up and there shall be full contact without air bubbles between the specimen and the surface of the culture medium. Coat the specimens with a layer (1 mm thick) of the algal sus
