1、BSI Standards PublicationBS EN 16421:2014Influence of materials on waterfor human consumption Enhancement of microbialgrowth (EMG)BS EN 16421:2014Incorporating corrigendum December 2015BS EN 16421:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 16421:2014.T
2、he purpose of this standard is to describe three European test methods that can be applied to determine the ability of nonmetallic materials to enhance microbial growth in drinking water.Each method uses different performance characteristics, which allows its use for specific materials or product ty
3、pes, but all three methods have limitations. For example, multi-layer pipes cannot currently be tested with the Method 1 and Method 3, and small components cannot easily be analysed using Method 2. Harmonized product standards will specify the particular method to follow, taking into account materia
4、l of construction and type of components. Pass/fail criteria is not included in this standard and will be specified by the individual Member State.At the time of publication, this standard is not currently included in the (UK) product testing to be used for materials in contact with drinking water a
5、s all the methodology in BS 6920, together with the pass/fail criteria, has not been replaced by European Standards. Studies to establish the degree of agreement of the results obtained using the three methods on a wide range of specific materials types will enable the establishment of comparative p
6、ass/fail criteria. To ensure the correct methodology is being used, refer to the National Approval schemes for the UK before undertaking any testing. Further information can be obtained from UK approvals bodies, such as the Drinking Water Inspectorate and the Water Regulations Advisory Scheme.The UK
7、 participation in its preparation was entrusted to Technical Committee EH/6, Effects of materials on water quality.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract
8、. Users are responsible for its correct application. The British Standards Institution 2015. Published by BSI Standards Limited 2015ISBN 978 0 580 92629 7ICS 07.100.20Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the author
9、ity of the Standards Policy and Strategy Committee on 31 January 2015.Amendments/corrigenda issued since publicationDate Text affected31 December 2015 Implementation of CEN Correction Notice 03 February 2015: Subclauses 5.5.2.2 and 5.6.9 modifiedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 1
10、6421 December 2014 ICS 07.100.20 English Version Influence of materials on water for human consumption - Enhancement of microbial growth (EMG) Influence des matriaux sur leau destine la consommation humaine - Stimulation de la croissance microbienne (SCM) Einfluss von Materialien auf Wasser fr den m
11、enschlichen Gebrauch - Frderung des mikrobiellen Wachstums This European Standard was approved by CEN on 25 October 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 witho
12、ut 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 other l
13、anguage 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, Den
14、mark, 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 COMMITTE
15、E 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 16421:2014 EEN 16421:2014 (E)
16、 2 Contents Page Foreword 6 Introduction .7 1 Scope 9 2 Normative references 9 3 Terms and definitions 10 4 Influence of materials on water for human consumption Enhancement of microbial growth (EMG) Method 1: Measured by Biomass Production Potential (BPP) measured by ATP 11 4.1 General . 11 4.2 Pri
17、nciple . 12 4.3 Apparatus 12 4.3.1 Requirements 12 4.3.2 Vessels, containers, stoppers and connectors and general laboratory apparatus . 12 4.3.3 Cleaning . 12 4.3.4 Gloves 12 4.3.5 Plates . 12 4.3.6 Test containers . 13 4.3.7 Test tubes 13 4.3.8 Incubator (or hot room) 13 4.3.9 Sterilisation ovens
18、13 4.3.10 Membrane filters . 13 4.3.11 Autoclave . 13 4.4 Reagents 13 4.4.1 Test water 13 4.4.2 Dilution water 14 4.4.3 Potassium dihydrogen orthophosphate solution . 14 4.4.4 Potassium-nitrate solution 14 4.4.5 Sodium acetate solution 15 4.4.6 Inoculum 15 4.4.7 Specifications for controls 15 4.5 Te
19、st samples and controls 16 4.5.1 Sampling, transport and storage of test samples . 16 4.5.2 Test piece preparation . 16 4.6 Pre-treatment of test pieces 17 4.6.1 General . 17 4.6.2 Flushing . 17 4.6.3 Stagnation . 17 4.6.4 Prewashing 17 4.7 Test procedure 17 4.7.1 Overview 17 4.7.2 ATP contamination
20、 . 18 4.7.3 Preparation of the test container 18 4.7.4 Test pieces 18 4.7.5 Incubation 18 4.7.6 Positive and negative controls 18 4.7.7 Changing of test water . 19 4.7.8 Measurement of ATP concentrations . 19 4.8 Calculation of test results 19 4.8.1 Biomass Production (BP) 19 4.9 Reproducibility and
21、 repeatability 22 BS EN 16421:2014EN 16421:2014 (E) 2 Contents Page Foreword 6 Introduction .7 1 Scope 9 2 Normative references 9 3 Terms and definitions 10 4 Influence of materials on water for human consumption Enhancement of microbial growth (EMG) Method 1: Measured by Biomass Production Potentia
22、l (BPP) measured by ATP 11 4.1 General . 11 4.2 Principle . 12 4.3 Apparatus 12 4.3.1 Requirements 12 4.3.2 Vessels, containers, stoppers and connectors and general laboratory apparatus . 12 4.3.3 Cleaning . 12 4.3.4 Gloves 12 4.3.5 Plates . 12 4.3.6 Test containers . 13 4.3.7 Test tubes 13 4.3.8 In
23、cubator (or hot room) 13 4.3.9 Sterilisation ovens 13 4.3.10 Membrane filters . 13 4.3.11 Autoclave . 13 4.4 Reagents 13 4.4.1 Test water 13 4.4.2 Dilution water 14 4.4.3 Potassium dihydrogen orthophosphate solution . 14 4.4.4 Potassium-nitrate solution 14 4.4.5 Sodium acetate solution 15 4.4.6 Inoc
24、ulum 15 4.4.7 Specifications for controls 15 4.5 Test samples and controls 16 4.5.1 Sampling, transport and storage of test samples . 16 4.5.2 Test piece preparation . 16 4.6 Pre-treatment of test pieces 17 4.6.1 General . 17 4.6.2 Flushing . 17 4.6.3 Stagnation . 17 4.6.4 Prewashing 17 4.7 Test pro
25、cedure 17 4.7.1 Overview 17 4.7.2 ATP contamination . 18 4.7.3 Preparation of the test container 18 4.7.4 Test pieces 18 4.7.5 Incubation 18 4.7.6 Positive and negative controls 18 4.7.7 Changing of test water . 19 4.7.8 Measurement of ATP concentrations . 19 4.8 Calculation of test results 19 4.8.1
26、 Biomass Production (BP) 19 4.9 Reproducibility and repeatability 22 EN 16421:2014 (E) 3 4.10 Test report 22 4.10.1 General . 22 4.10.2 Material/Product information 22 4.10.3 Information on the test procedure . 23 4.10.4 Test results 23 5 Influence of materials on water for human consumption Enhance
27、ment of microbial growth (EMG) Method 2: Measured by biofilm volume 23 5.1 General . 23 5.2 Principle 24 5.3 Basic requirements . 24 5.4 Apparatus and reagents . 24 5.4.1 Disinfectant 24 5.4.2 Test water . 24 5.4.3 Specifications for controls . 25 5.4.4 Test container and test module for pipes and h
28、oses 25 5.4.5 Scraper . 25 5.4.6 Centrifuge . 25 5.4.7 Glassware for centrifugation 25 5.4.8 Flowmeter . 25 5.5 Test samples and test pieces . 25 5.5.1 Sampling, transport and storage . 25 5.5.2 Preparation of test pieces. 26 5.6 Test procedure . 27 5.6.1 General . 27 5.6.2 Test procedure in test co
29、ntainer 27 5.6.3 Test procedure in test module for pipes and hoses 27 5.6.4 Preparation of the test container or test module for pipes and hoses 28 5.6.5 Pre-treatment of test pieces . 28 5.6.6 Removal of test pieces 28 5.6.7 Examination of test pieces . 28 5.6.8 Re-exposure after harvesting . 28 5.
30、6.9 Centrifugation and determination of biofilm 28 5.6.10 Examination of biofilm 29 5.6.11 Test periods . 29 5.7 Calculation of test results. 30 5.8 Evaluation of satisfactory test performance 31 5.9 Test report 32 5.9.1 General . 32 5.9.2 Material/Product information 32 5.9.3 Information on the tes
31、t procedure . 33 5.9.4 Test results 33 6 Influence of materials on water for human consumption Enhancement of microbial growth (EMG) Method 3: Measured by mean dissolved oxygen depletion 34 6.1 General . 34 6.2 Principle 34 6.3 Test premises 34 6.3.1 General . 34 6.3.2 Safety 34 6.4 Apparatus and re
32、agents . 34 6.4.1 General . 34 6.4.2 Inoculum water 34 6.4.3 Test water . 35 6.4.4 Test containers 36 6.4.5 Dissolved oxygen meter . 37 6.4.6 30 C Incubator/chamber 37 6.4.7 Polyethylene bags . 37 6.4.8 Glass plates . 37 BS EN 16421:2014EN 16421:2014 (E) 4 6.4.9 Brass couplings 37 6.4.10 Special gla
33、ss plates 37 6.5 Preparation of test pieces 37 6.5.1 General . 37 6.5.2 Nature of test pieces 37 6.6 Test piece and test container size 37 6.6.1 General . 37 6.6.2 Test sample storage . 38 6.6.3 Special requirements . 38 6.6.4 Specifications for controls 42 6.6.5 Test water control . 42 6.7 Test pro
34、cedures 42 6.7.1 Preparation of the test container 42 6.7.2 Incubation 43 6.7.3 Changing the test water . 43 6.7.4 Measurement of dissolved oxygen . 43 6.8 Calculation of results . 44 6.8.1 Calculation of the mean dissolved oxygen difference (MDOD) . 44 6.8.2 Validation of results 44 6.8.3 Repeatabi
35、lity and reproducibility 45 6.9 Test report . 45 6.9.1 General . 45 6.9.2 Material/Product information . 45 6.9.3 Information on the test procedure 47 6.9.4 Results . 47 Annex A (normative) Method 1: Measured by Biomass Production Potential (BPP) measured by ATP Assessment of the suitability of test
36、 water for its use in the BPP test (biological stability and the need for addition of trace elements) 48 Annex B (normative) Method 1: Measured by Biomass Production Potential (BPP) measured by ATP Protocol of ATP analysis 50 B.1 Introduction . 50 B.2 Apparatus and glassware 50 B.3 Reagents and auxi
37、liary substances 50 B.3.1 General . 50 B.3.2 Sterile water . 50 B.3.3 ATP solutions for standard dilutions and calibration curves 50 B.3.4 Luciferin/luciferase (enzyme) solution . 51 B.3.5 ATP extraction reagent 52 B.4 Sampling 52 B.5 Procedure 52 B.5.1 General . 52 B.5.2 Blank water 52 B.5.3 Contro
38、l dilutions . 52 B.5.4 Samples . 52 B.6 Expression of results . 52 B.6.1 Water samples . 52 B.6.2 Material samples . 53 B.7 Test record 53 BS EN 16421:2014EN 16421:2014 (E) 4 6.4.9 Brass couplings 37 6.4.10 Special glass plates 37 6.5 Preparation of test pieces 37 6.5.1 General . 37 6.5.2 Nature of
39、test pieces 37 6.6 Test piece and test container size 37 6.6.1 General . 37 6.6.2 Test sample storage . 38 6.6.3 Special requirements . 38 6.6.4 Specifications for controls 42 6.6.5 Test water control . 42 6.7 Test procedures 42 6.7.1 Preparation of the test container 42 6.7.2 Incubation 43 6.7.3 Ch
40、anging the test water . 43 6.7.4 Measurement of dissolved oxygen . 43 6.8 Calculation of results . 44 6.8.1 Calculation of the mean dissolved oxygen difference (MDOD) . 44 6.8.2 Validation of results 44 6.8.3 Repeatability and reproducibility 45 6.9 Test report . 45 6.9.1 General . 45 6.9.2 Material
41、/Product information . 45 6.9.3 Information on the test procedure 47 6.9.4 Results . 47 Annex A (normative) Method 1: Measured by Biomass Production Potential (BPP) measured by ATP Assessment of the suitability of test water for its use in the BPP test (biological stability and the need for addition
42、 of trace elements) 48 Annex B (normative) Method 1: Measured by Biomass Production Potential (BPP) measured by ATP Protocol of ATP analysis 50 B.1 Introduction . 50 B.2 Apparatus and glassware 50 B.3 Reagents and auxiliary substances 50 B.3.1 General . 50 B.3.2 Sterile water . 50 B.3.3 ATP solution
43、s for standard dilutions and calibration curves 50 B.3.4 Luciferin/luciferase (enzyme) solution . 51 B.3.5 ATP extraction reagent 52 B.4 Sampling 52 B.5 Procedure 52 B.5.1 General . 52 B.5.2 Blank water 52 B.5.3 Control dilutions . 52 B.5.4 Samples . 52 B.6 Expression of results . 52 B.6.1 Water sam
44、ples . 52 B.6.2 Material samples . 53 B.7 Test record 53 EN 16421:2014 (E) 5 B.8 Quality control of calibration curves . 53 Annex C (informative) Method 1: Measured by Biomass Production Potential (BPP) measured by ATP - ATP and its measurement 55 Annex D (informative) Method 1: Measured by Biomass
45、Production Potential (BPP) measured by ATP Evaluating procedures for removal of attached biomass 56 D.1 Introduction 56 D.2 Procedures for removal of Attached Biomass . 56 D.2.1 General . 56 D.2.2 Swabbing 56 D.2.3 Low energy sonication (LES) . 56 D.2.4 High energy sonication (HES) 56 D.3 Effectiven
46、ess of removal procedure . 57 Annex E (informative) Method 1: Measured by Biomass Production Potential (BPP) measured by ATP 58 Annex F (informative) Method 2: Measured by biofilm volume 59 Annex G (informative) Method 2: Measured by biofilm volume . 60 Annex H (informative) Method 3: Measured by me
47、an dissolved oxygen depletion . 63 Bibliography 65 BS EN 16421:2014EN 16421:2014 (E) 6 Foreword This document (EN 16421:2014) has been prepared by Technical Committee CEN/TC 164 “Water supply”, the secretariat of which is held by AFNOR. This European Standard shall be given the status of a national
48、standard, either by publication of an identical text or by endorsement, at the latest by June 2015, and conflicting national standards shall be withdrawn at the latest by June 2015. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights.
49、CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. 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, 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, Spai
