1、BSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06Soil quality - Contact test for solid samples using the dehydrogenase activity of Arthrobacter globiformisBS EN ISO 18187:2018EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 18187 February 2018 ICS 13.080.30 En
2、glish Version Soil quality - Contact test for solid samples using the dehydrogenase activity of Arthrobacter globiformis (ISO 18187:2016) Qualit du sol - Essai contact pour chantillons solides utilisant lactivit dshydrognase de Arthrobacter globiformis (ISO 18187:2016) Bodenbeschaffenheit - Feststof
3、fkontakttest unter Verwendung der Dehydrogenaseaktivitt von Arthrobacter globiformis (ISO 18187:2016) This European Standard was approved by CEN on 14 February 2018. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Stan
4、dard the status of a national standard 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 (Englis
5、h, 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-CENELEC Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bu
6、lgaria, 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, Serbia, Slovakia, Slovenia, Spain, Sweden, Switz
7、erland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels 2018 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN nat
8、ional Members. Ref. No. EN ISO 18187:2018 ENational forewordThis British Standard is the UK implementation of EN ISO 18187:2018. It is identical to ISO 18187:2016. It supersedes BS ISO 18187:2016, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee EH/4, S
9、oil 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. Users are responsible for its correct application. The British Standards Institution 2018 Published b
10、y BSI Standards Limited 2018ISBN 978 0 580 98304 7ICS 13.080.30Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 30 April 2016.Amendments/corrigenda issued since p
11、ublicationDate Text affected31 March 2018 This corrigendum renumbers BS ISO 18187:2016 as BS EN ISO 18187:2018BRITISH STANDARDBS EN ISO 18187:2018EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 18187 February 2018 ICS 13.080.30 English Version Soil quality - Contact test for solid samples u
12、sing the dehydrogenase activity of Arthrobacter globiformis (ISO 18187:2016) Qualit du sol - Essai contact pour chantillons solides utilisant lactivit dshydrognase de Arthrobacter globiformis (ISO 18187:2016) Bodenbeschaffenheit - Feststoffkontakttest unter Verwendung der Dehydrogenaseaktivitt von A
13、rthrobacter globiformis (ISO 18187:2016) This European Standard was approved by CEN on 14 February 2018. 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
14、. 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 language made by t
15、ranslation 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, Denmark, Estonia, Fi
16、nland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STA
17、NDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels 2018 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 18187:2018 EBS EN ISO 18187:20
18、18EN ISO 18187:2018 (E) 3 European foreword The text of ISO 18187:2016 has been prepared by Technical Committee ISO/TC 190 “Soil quality” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 18187:2018 by Technical Committee CEN/TC 444 “Test methods for envir
19、onmental characterization of solid matrices” the secretariat of which is held by NEN. 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 August 2018, and conflicting national standards shall be withd
20、rawn at the latest by August 2018. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN shall not be held responsible for identifying any or all such patent rights. According to the CEN-CENELEC Internal Regulations, the national st
21、andards 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,
22、 Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 18187:2016 has been approved by CEN as EN ISO 18187:2018 without any modification. BS EN ISO 18187:2018ISO 181
23、87:2016(E)Foreword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 45 Reagents and material 45.1 Test organisms 45.2 Control substrates 45.2.1 General 45.2.2 Control for soils . 45.2.3 Control for waste material . 55.3 Test substrates 55.4 Chemicals . 66 App
24、aratus . 87 Procedure. 97.1 Preparation of dilutions 97.2 Reference substance and positive control preparation . 97.3 Contact test procedure . 97.3.1 General 97.3.2 Aeration 107.3.3 Deactivation 107.3.4 Preparation of the inoculum . 117.3.5 Incubation and fluorescence measurement 117.4 Interferences
25、 118 Calculation and expression of results 128.1 Calculation . 128.1.1 Relative fluorescence 128.1.2 Determining the percentage of inhibition 128.2 Expression of results . 129 Validity of the test .1310 Statistical analysis 1311 Test report 14Annex A (informative) Results on the ring test .15Annex B
26、 (informative) Preparation of test organisms 21Annex C (informative) Testing chemical substances .23Bibliography .25 ISO 2016 All rights reserved iiiContents PageBS EN ISO 18187:2018ISO 18187:2016(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of nationa
27、l 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 that committee. Internation
28、al 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.The procedures used to develop this document and those intended f
29、or its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.o
30、rg/directives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be
31、 in the Introduction and/or on the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expressions
32、related to conformity assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 4, Biologic
33、al methods.iv ISO 2016 All rights reservedBS EN ISO 18187:2018ISO 18187:2016(E)IntroductionThis International Standard describes the miniaturized solid contact assay with Arthrobacter globiformis that allows the preliminary assessment of solid material (i.e. soil and waste materials) within 6 h. The
34、 principle of the assay relies on dehydrogenase activity inhibition of an added test organism, caused by bioavailable toxic substances in soil and waste samples. This is an ecologically relevant assay as far as it uses a ubiquitous soil bacteria species with high affinity to surfaces166which dehydro
35、genases are involved in different biological mechanisms withstanding bacteria integrity (e.g. respiratory chains). Moreover, it has been noticed that this parameter (dehydrogenase activity inhibition) is quite sensitive to different toxic substances.19101415Overall, this assay is non-labour-intensiv
36、e, rapid, cost-effective and sensitive, providing results that improve the physical and chemical assessment of natural samples while allowing a quick indication of their biological effects.The miniaturized solid contact assay is based on the solid contact assay established by Reference 7.This Intern
37、ational Standard is also based on Reference 23.The results of an interlaboratory trial towards the evaluation of test variability to assess different waste and soil samples, as well as chemicals, are presented in Annex A. ISO 2016 All rights reserved vBS EN ISO 18187:2018BS EN ISO 18187:2018Soil qua
38、lity Contact test for solid samples using the dehydrogenase activity of Arthrobacter globiformis1 ScopeThis International Standard specifies a rapid method for assessing solid samples in an aerobic suspension, by determining the inhibition of dehydrogenase activity of Arthrobacter globiformis using
39、the redox dye resazurin.It is applicable for assessing the effect of water-soluble and solid matter bounded non-volatile contaminants of natural samples, such as soils and waste materials. The test yields a result within 6 h and can therefore be used for screening potentially contaminated material.2
40、 Normative referencesThe 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 (including any amendme
41、nts) applies.ISO 5667-15, Water quality Sampling Part 15: Guidance on the preservation and handling of sludge and sediment samplesISO 10381-6, Soil quality Sampling Part 6: Guidance on the collection, handling and storage of soil under aerobic conditions for the assessment of microbiological process
42、es, biomass and diversity in the laboratoryCEN/TR 15310-1, Characterization of waste Sampling of waste materials Part 1: Guidance on selection and application of criteria for sampling under various conditionsEN 14735, Characterization of waste Preparation of waste samples for ecotoxicity tests3 Term
43、s and definitionsFor the purposes of this document, the following terms and definitions apply.3.1contact timeexposure period of the bacteria to a suspension of solid matter3.2negative controlsample of a control substrate (3.6) with a mixture of known solutions distilled water, medium B or inoculum (
44、3.12).Note 1 to entry: It is used to standardize the analysis.3.3positive controlsample of a control substrate (3.6) with a mixture of known solutions distilled water, medium B or inoculum (3.12) and a reference substanceNote 1 to entry: It is used to check the sensitivity of the test organism.INTER
45、NATIONAL STANDARD ISO 18187:2016(E) ISO 2016 All rights reserved 1BS EN ISO 18187:2018ISO 18187:2016(E)3.4blank Ablank, which sets the own fluorescence of the substrate after being deactivatedNote 1 to entry: Blank is not added with bacteria.3.5blank Bblank, which sets the natural fluorescence of th
46、e substrate without being deactivatedNote 1 to entry: Blank is not added with bacteria.3.6control substratereference or standard substrate used as a control and as medium (3.13) for preparing dilution/concentration series with test substrates (3.7) or a reference substanceEXAMPLE Quartz sand or LUFA
47、 standard soil type 2.2.3.7test substratenatural or artificial substrate that is naturally contaminated or spiked with a test chemicalNote 1 to entry: The test substrate is the test material (3.8) after being prepared for testing (e.g. sieved) and/or diluted with a control substrate (3.6).3.8test ma
48、terialoriginal sample of soil or waste material without any changes (e.g. sieving)3.9dehydrogenase activityactivity of hydrogen-abstracting enzymes which are involved in many energy and biosynthesis metabolic processes (e.g. the respiratory chain) and which require cell integrity to be produced Note
49、 1 to entry: These enzymes can reduce resazurin into resorufin in the extracellular environment.6Note 2 to entry: See Reference 21.3.10effect concentration for x % effectECxconcentration (mass fraction) of a test substance or sample that causes x % of an effect on a given endpoint within a given exposure period when compared with a controlEXAMPLE An EC50 is a concentration estimated to cause an effect on a test end point in 50 % of an exposed population over a defined exposure period.Note 1 to entry: The ECx
