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1、BRITISH STANDARD BS ISO 16869:2001 Plastics Assessment of the effectiveness of fungistatic compounds in plastics formulations ICS 83.080.01 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrol
2、led Copy, (c) BSI BS ISO 16869:2001 This British Standard, having been prepared under the direction of the Sector Policy and Strategy Committee for Materials and Chemicals, was published under the authority of the Standards Policy and Strategy Committee on 16 January 2002 BSI 16 January 2002 ISBN 0
3、580 38857 3 National foreword This British Standard reproduces verbatim ISO 16869:2001 and implements it as the UK national standard. The UK participation in its preparation was entrusted to Technical Committee PRI/21, Plastics test methods, which has the responsibility to: A list of organizations r
4、epresented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards Corresp
5、ondence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not
6、of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European development
7、s and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the ISO title page, pages ii to v, a blank page, pages 1 to 9 and a back cover. The BSI copyright date displayed in this document indicates when the document was last issued. Amendments is
8、sued since publication Amd. No. DateComments Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI Reference number ISO 16869:2001(E) ISO1002 INTERNATIONAL STANDARD ISO 16869 First edition 2001-11-15 Plastics Assessment of the effectiveness of fun
9、gistatic compounds in plastics formulations Plastiques valuation de lefficacit des composs fongistatiques dans les formulations de plastiques Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ii Licensed Copy: sheffieldun sheffieldun, na, Mon
10、Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ISO 16869:2001(E) iii Contents Page Foreword.iv Introduction.v 1 Scope1 2 Normative references1 3 Terms and definitions .1 4 Principle2 5 Apparatus and materials.2 6 Test specimens5 7 Preparation of specimens.5 8 Procedure.6 9 Assessment of
11、fungal growth 7 10 Expression of results7 11 Precision and bias.8 12 Test report8 Bibliography9 Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ISO 16869:2001(E) iv Foreword ISO (the International Organization for Standardization) is a world
12、wide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on t
13、hat committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in
14、 accordance with the rules given in the ISO/IEC Directives, Part 3. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard
15、requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this International Standard may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. International S
16、tandard ISO 16869 was prepared by Technical Committee ISO/TC 61, Plastics, Subcommittee SC 6, Ageing, chemical and environmental resistance. Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ISO 16869:2001(E) v Introduction It is a well known
17、phenomenon that plasticizers as well as other ingredients in plastics formulations can be attacked by bacteria, yeasts and fungi, the latter being the most important deteriogens. Microbial attack results in a reduction of the quality of the plastic, causing embrittlement as well as discoloration. Th
18、is deterioration is of economic importance. The prevention of fungal attack can be achieved by the incorporation of a fungistatic compound into the formulation. The function of this fungistat is to inhibit the growth of any fungi present on the surface of the plastic product. The method described in
19、 this International Standard determines the effectiveness of fungistatic compounds incorporated into the plastic against the fungi used in the test. Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI blank Licensed Copy: sheffieldun sheffieldun
20、, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI INTERNATIONAL STANDARD ISO 16869:2001(E) 1 Plastics Assessment of the effectiveness of fungistatic compounds in plastics formulations WARNING Handling and manipulation of microorganisms that are potentially hazardous requires a hig
21、h degree of technical competence and may be subject to current national legislation and regulations. Only personnel trained in microbiological techniques should carry out such tests. Codes of practice for disinfection, sterilization and personal hygiene must be strictly observed. It is recommended t
22、hat workers consult IEC 60068-2-10:1988, appendix A “Danger to personnel”, and ISO 7218:1996, Microbiology of food and animal feeding stuffs General rules for microbiological examinations. 1 Scope This International Standard specifies a method for determining the effectiveness of fungistatic compoun
23、ds in protecting susceptible ingredients like plasticizers, stabilizers, etc., in plastics formulations. The method demonstrates whether or not a plastic product is actively protected against fungal attack. The evaluation is by visual examination. The test is applicable to any articles made of plast
24、ic that are in the form of films or plates no thicker than 10 mm. In addition, porous materials such as plastic foams may be tested provided that they are in the above-mentioned form. In contrast to ISO 846, the test films are not sprayed with a fungal spore suspension but covered with a layer of te
25、st agar containing spores. It has been found that this leads to a better distribution of the spores as well as providing a good supply of water necessary for spore germination on plastic surfaces that are normally hydrophobic. 2 Normative references The following normative documents contain provisio
26、ns which, through reference in this text, constitute provisions of this International Standard. For dated references, subsequent amendments to, or revisions of, any of these publications do not apply. However, parties to agreements based on this International Standard are encouraged to investigate t
27、he possibility of applying the most recent editions of the normative documents indicated below. For undated references, the latest edition of the normative document referred to applies. Members of ISO and IEC maintain registers of currently valid International Standards. ISO 291:1997, Plastics Stand
28、ard atmospheres for conditioning and testing EN 554:1994, Sterilization of medical devices Validation and routine control of sterilization by moist heat 3 Terms and definitions For the purposes of this International Standard, the following terms and definitions apply. 3.1 plastic susceptible to fung
29、al attack plastic material that contains in its formulation one or more nutrients that support fungal growth Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ISO 16869:2001(E) 2 3.2 fungistat a compound that prevents fungal growth on a materi
30、al that is normally susceptible to fungal attack 4 Principle Test specimens are exposed to a suspension of mixed fungal spores. The spores are applied to the surface of the test specimen in a thin layer of an agar medium without an added carbon source. In this way, uniform distribution of the spores
31、 is achieved as well as an optimum supply of water. The absence of fungistatic agents in the plastic material will lead to germination of the fungal spores and initial growth. When the ingredients in the material are susceptible to fungal attack and no active fungistat is included in the formulation
32、, further growth and sporulation will occur over and around the test specimen. The presence of an active fungistat in the material will lead to suppression of spore germination and initial growth in the area over and around the test specimen. Fungistatic agents can migrate into the agar around the t
33、est specimen, thereby suppressing germination and appearing to give an increased zone of inhibition. Although not relevant to the interpretation of the test results, the inhibition zone can be an indication of the behaviour of the fungistat under test. 5 Apparatus and materials 5.1 Apparatus Sterili
34、ze all glassware and all parts of the rest of the apparatus that will come into contact with the culture media and/or reagents (except those which are supplied sterile) by one of the following methods: method A: autoclave (see 5.1.2) at 121 C for a minimum of 15 min; method B: use a dry-heat sterili
35、zer (see 5.1.2) at 180 C for at least 30 min, at 170 C for at least 1 h, or at 160 C for at least 2 h; method C: use a membrane-filtration system of pore size 0,45 m. 5.1.1 Incubator, maintained at 24 C 1 C. 5.1.2 Sterilization apparatus: 5.1.2.1 For moist-heat sterilization, an autoclave complying
36、with the requirements of EN 554. 5.1.2.2 For dry-heat sterilization, a hot-air oven maintained at one of the temperatures specified in method B above. 5.1.2.3 For membrane-filtration sterilization, a membrane-filtration apparatus, of pore size as specified in method C above. 5.1.3 Analytical balance
37、, accurate to 0,1 mg. 5.1.4 Laboratory centrifuge, speed 2 000 rpm to 5 000 rpm. 5.1.5 Counting chamber (for direct counting with the help of a microscope). 5.1.6 Microscope, magnification 100. 5.1.7 pH-meter, having an accuracy of 0,1 pH-units, capable of temperature correction. Licensed Copy: shef
38、fieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ISO 16869:2001(E) 3 5.1.8 Vortex shaker, operating at 2 000 rpm to 2 500 rpm. 5.1.9 Containers: test tubes, flasks or bottles of suitable capacity. 5.1.10 Petri dishes, 90 mm to 100 mm in diameter and at least 15
39、 mm deep. 5.1.11 Graduated pipettes, with nominal capacities of 1,0 ml and 15,0 ml. Calibrated automatic pipettes may be used. 5.1.12 Graduated measuring cylinder, minimum capacity 30 ml. 5.1.13 Glass beads, diameter 3 mm to 5 mm. 5.2 Culture media and reagents All reagents shall be of analytical gr
40、ade and/or of a grade appropriate for microbiological purposes. 5.2.1 Water Any water used shall be distilled or deionized and have a conductivity of 1 S/cm. 5.2.2 Malt-extract agar (MEA) Malt extract 30,0 g Soya peptone 3,0 g Agar-agar 15,0 g Water (5.2.1) to make up to 1 000 ml Sterilize in the au
41、toclave (see 5.1.2). After sterilization, the pH of the medium shall be 7,0 0,2. 5.2.3 Chaetomium agar NaNO3 2,0 g MgSO47H2O 0,5 g KCl 0,5 g Fe2(SO4)3H2O 0,01 g KH2PO4 0,14 g K2HPO4 1,20 g Agar-agar 15,0 g Yeast extract 0,02 g Microcellulose 20,0 g or Carboxymethyl-cellulose (Na salt) 10,0 g Water (
42、5.2.1) to make up to 1 000 ml Sterilize in the autoclave (see 5.1.2). After sterilization, the pH of the medium shall be 7,2 0,2. Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ISO 16869:2001(E) 4 5.2.4 Wetting agent Prepare a 5 % (m/V) sto
43、ck solution of polysorbate 80 (polyoxyethylenesorbitane monooleate) in water. To harvest the spores, dilute the stock solution with water to 0,05 % (m/V). 5.2.5 Stock solution for nutrient-salt solution and agar NaCl 0,5 g FeSO47H2O 0,2 g ZnSO47H2O 0,2 g MnSO41H2O 0,06 g Water (5.2.1) to make up to
44、1 000 ml Before storage for a lengthy period, the stock solution shall be sterilized by membrane filtration. 5.2.6 Nutrient-salt solution KH2PO4 2,62 g Na2HPO42H2O 0,2 g MgSO47H2O 0,7 g NH4NO3 1,0 g Stock solution (5.2.5) 10 ml Water (5.2.1) to make up to 1 000 ml Sterilize in the autoclave (see 5.1
45、.2). After sterilization, the pH of the medium shall be 5,5 0,2. 5.2.7 Nutrient-salt agar KH2PO4 2,62 g Na2HPO42H2O 0,20 g MgSO47H2O 0,70 g NH4NO3 1,0 g Agar-agar 15,0 g Stock solution (5.2.5) 10 ml Water (5.2.1) to make up to 1 000 ml Heat the prepared medium till the agar-agar is molten, then meas
46、ure the temperature and adjust the pH to 5,5 0,1. Sterilize in an autoclave (see 5.1.2). Agar plates shall be freshly prepared when no validated method for long-term storage (longer than 3 days) is available. The sterilized solutions may be stored for up to 3 months. The storage conditions shall pre
47、clude any evaporation occurring. Licensed Copy: sheffieldun sheffieldun, na, Mon Nov 27 08:14:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ISO 16869:2001(E) 5 5.3 Organisms and cultivation 5.3.1 Test organisms 5.3.1.1 Aspergillus niger ATCC 6275 5.3.1.2 Chaetomium globosum ATCC 6205 5.3.1.3 Paecilo
48、myces variotii CBS 628.66 5.3.1.4 Penicillium funiculosum ATCC 9644 5.3.1.5 Trichoderma longibrachiatum ATCC 13631 The test fungi shall be obtained from a national culture collection (e.g. ATCC = American Type Culture Collection, USA; CBS = Centraalbureau voor Schimmelcultures, NL). If there are par
49、ticular reasons for doing so, and by agreement between the interested parties, other fungi may be used. In any case, all strains used shall be listed in the test report. 5.3.2 Culture conditions Cultivation of strains 5.3.1.1 and 5.3.1.3 to 5.3.1.5 shall be on a malt-extract agar (5.2.2) slant in a test tube at 24 C 1 C for 14 to 21 days. Cultivation of strain 5.3.1.2 shall be on Chaetomium agar (5.2.3) at 24 C 1 C for 14 to 21 days. Stock cultures may be kept on agar slants or, preferably, freeze-dried or frozen. 6 Test speci
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