1、June 2012 Translation by DIN-Sprachendienst.English price group 14No 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).ICS 1
2、3.060.70!$|“1896196www.din.deDDIN EN ISO 8692Water quality Fresh water algal growth inhibition test with unicellular green algae(ISO 8692:2012)English translation of DIN EN ISO 8692:2012-06Wasserbeschaffenheit Swasseralgen-Wachstumshemmtest mit einzelligen Grnalgen (ISO 8692:2012)Englische bersetzun
3、g von DIN EN ISO 8692:2012-06Qualit de leau Essai dinhibition de la croissance des algues deau douce avec des algues vertesunicellulaires (ISO 8692:2012)Traduction anglaise de DIN EN ISO 8692:2012-06SupersedesDIN EN ISO 8692:2005-01www.beuth.deIn case of doubt, the German-language original shall be
4、considered authoritative.Document comprises 28 pages05.12 DIN EN ISO 8692:2012-06 2 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee ISO/TC 147 “Water quality” in collaboration with Technical Committee CEN/TC 230 “Water analysis” (Secret
5、ariat: DIN, Germany). The responsible German body involved in its preparation was the Normenausschuss Wasserwesen (Water Practice Standards Committee), Working Committee NA 119-01-03 AA Wasseruntersuchung, Working Group NA 119-01-03-05-01 AK Biotests. This standard is part of the series Deutsche Ein
6、heitsverfahren zur Wasser-, Abwasser- und Schlammuntersuchung Testverfahren mit Wasserorganismen (Gruppe L) (German standard methods for the examination of water, waste water and sludge Test methods using water organisms (group L) and describes method L 9. Designation of the method: Fresh water alga
7、l growth inhibition test with unicellular green algae (L 9): Method DIN EN ISO 8692 L 9 This standard has been published to implement the Water Framework Directive (WFD), Directive 2000/60/EC of the European Parliament and of the Council of 23 October 2000 establishing a framework for Community acti
8、on in the field of water policy.*)The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 5667-16 DIN EN ISO 5667-16 ISO 15088 DIN EN ISO 15088 Amendments This standard differs from DIN EN ISO 8692:2005-01 as follows: a) a definition of the ter
9、m “lowest ineffective dilution” (LID) has been included (3.3); b) Annex B specifying an alternative test procedure with algae from algal beads, with direct measurement of algal growth in spectrophotometric cells has been included; c) a new Annex C covering the procedure for immobilization of algae i
10、n alginate beads has been included (see 5.1); d) the standard has been editorially revised. Previous editions DIN 38412-9: 1989-05 DIN EN 28692: 1993-04 DIN EN ISO 8692: 2005-01 *)Registered in the DITR database of DIN Software GmbH, obtainable from: Beuth Verlag GmbH, 10772 Berlin. DIN EN ISO 8692:
11、2012-06 3 Expert assistance and specialized laboratories will be required to perform the analysis described in this standard. Existing safety regulations are to be taken into account. Depending on the objective of the analysis, a check shall be made on a case-by-case basis as to whether and to what
12、extent additional conditions will have to be specified. German standard methods for the examination of water, waste water and sludge Standard methods published as DIN Standards are obtainable from Beuth Verlag GmbH, either individually or grouped in volumes. The standard methods included in the loos
13、e-leaf publication entitled Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlammuntersuchung will continue to be published jointly by Wiley-VCH Verlag and Beuth Verlag GmbH. All standard methods relevant to the Abwasserverordnung (Waste Water Regulation) (AbwV)*) included in the new Regulat
14、ion on Section 57, paragraph 1, number 1, of the Gesetz zur Ordnung des Wasserhaushaltes (German Water Management Act*) concerning Anforderungen an das Einleiten von Abwasser in Gewsser together with the Abwasserverordnung and the Gesetz zur Ordnung des Wasserhaushalts have been published by Beuth V
15、erlag GmbH as a loose-leaf collection Analysenverfahren in der Abwasserverordnung Rechtsvorschriften und Normen (Supplements). Standards or draft standards bearing the group title “German standard methods for the examination of water, waste water and sludge” are classified under the following catego
16、ries (main titles): General information (group A) (DIN 38402) Sensory analysis (group B) (DIN 38403) Physical and physicochemical parameters (group C) (DIN 38404) Anions (group D) (DIN 38405) Cations (group E) (DIN 38406) Substance group analysis (group F) (DIN 38407) Gaseous constituents (group G)
17、(DIN 38408) Parameters characterizing effects and substances (group H) (DIN 38409) Biological-ecological methods of analysis (group M) (DIN 38410) Microbiological methods (group K) (DIN 38411) Test methods using water organisms (group L) (DIN 38412) Individual constituents (group P) (DIN 38413) Slud
18、ge and sediments (group S) (DIN 38414) Bio-assays with microorganisms (group T) (DIN 38415) In addition to the methods described in the DIN 38402 to DIN 38415 series of standards, there are a number of European and International Standards available as DIN EN, DIN EN ISO and DIN ISO Standards, which
19、also form part of the collection of German standard methods. Information on Parts of these series of standards that have already been published can be obtained from the offices of the Normenausschuss Wasserwesen, telephone +49 30 2601-2448, or from Beuth Verlag GmbH, Burggrafenstr. 6, 10787 Berlin.
20、*)Registered in the DITR database of DIN Software GmbH, obtainable from: Beuth Verlag GmbH, 10772 Berlin. DIN EN ISO 8692:2012-06 4 National Annex NA (informative) Bibliography DIN EN ISO 5667-16, Water quality Sampling Part 16: Guidance on biotesting of samples DIN EN ISO 15088, Water quality Deter
21、mination of the acute toxicity of waste water to zebrafish eggs (Danio rerio) EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 8692 February 2012 ICS 13.060.70 Supersedes EN ISO 8692:2004English Version Water quality - Fresh water algal growth inhibition test with unicellular green algae (IS
22、O 8692:2012) Qualit de leau - Essai dinhibition de la croissance des algues deau douce avec des algues vertes unicellulaires (ISO 8692:2012) Wasserbeschaffenheit - Swasseralgen-Wachstumshemmtest mit einzelligen Grnalgen 8692:2012) This European Standard was approved by CEN on 14 February 2012. CEN m
23、embers 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 on applic
24、ation 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-CENELEC Mana
25、gement 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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Nether
26、lands, 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 Management Centre: Avenue Marnix 17, B-1000 Brussels 2012 CEN All rights of exploi
27、tation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 8692:2012: E(ISODIN EN ISO 8692:2012-06 2 EN ISO 8692:2012 (E) Contents Page Foreword .3 1 Scope .4 2 Normative reference .4 3 Terms and definitions 4 4 Principle .5 5 Reagents and media .5 6 Apparatus 7
28、 7 Procedure 8 7.1 Preparation of growth medium .8 7.2 Preparation of pre- culture and inoculum 8 7.3 Choice of test sample concentrations 8 7.4 Preparation of test sample and stock solutions .8 7.5 Preparation of test and control batches .9 7.6 Incubation 9 7.7 Measurements 10 8 Validity criteria .
29、 10 9 Calculation 10 9.1 Plotting of growth curves . 10 9.2 Calculation of percentage inhibition . 11 9.3 Determination of ErCx(e.g. ErC10and ErC50) 11 10 Expression of results 11 11 Interpretation of results 12 12 Precision . 12 13 Test report 12 Annex A (normative) Rapid screening of waste water a
30、lgal growth inhibition 14 Annex B (normative) Test procedure with algae from algal beads, with direct measurement of algal growth in spectrophotometric cells . 17 Annex C (normative) Procedure for immobilization of algae in alginate beads . 22 Bibliography 24 EN ISO 8692:2012 (E) 3 Foreword This doc
31、ument (EN ISO 8692:2012) has been prepared by Technical Committee ISO/TC 147 “Water quality” in collaboration with Technical Committee CEN/TC 230 “Water analysis” the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by publication o
32、f an identical text or by endorsement, at the latest by August 2012, and conflicting national standards shall be withdrawn at the latest by August 2012. 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
33、be held responsible for identifying any or all such patent rights. This document supersedes EN ISO 8692:2004. 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,
34、Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, 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. Endorsement noti
35、ce The text of ISO 8692:2012 has been approved by CEN as a EN ISO 8692:2012 without any modification. DIN EN ISO 8692:2012-06 WARNING Persons using this International Standard should be familiar with normal laboratory practice. This International Standard does not purport to address all of the safet
36、y problems, if any, associated with its use. It is the responsibility of the user to establish appropriate safety and health practices and to ensure compliance with any national regulatory conditions.IMPORTANT It is absolutely essential that tests conducted in accordance with this International Stan
37、dard be carried out by suitably qualified staff.1 ScopeThis International Standard specifies a method for the determination of the growth inhibition of unicellular green algae by substances and mixtures contained in water or by waste water. This method is applicable for substances that are easily so
38、luble in water.With modifications to this method, as specified in ISO 14442 and ISO 5667-16, the inhibitory effects of poorly soluble organic and inorganic materials, volatile compounds, heavy metals and waste water can be tested.A rapid algal growth inhibition screening test for waste water is desc
39、ribed in Annex A.An alternative test procedure with algae from algal beads, with direct measurement of algal growth in spectrophotometric cells, is described in Annex B.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensab
40、le 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.ISO 5667-16, Water quality Sampling Part 16: Guidance on biotesting of samplesISO/TR 11044, Water quality Scientific
41、and technical aspects of batch algae growth inhibition testsISO 14442, Water quality Guidelines for algal growth inhibition tests with poorly soluble materials, volatile compounds, metals and waste waterISO/TS 20281, Water quality Guidance on statistical interpretation of ecotoxicity data3 Terms and
42、 definitionsFor the purposes of this document, the following terms and definitions apply.3.1cell densitynnumber of cells per volume of mediumNOTE Cell density is expressed in cells per millilitre.EN ISO 8692:2012 (E) 4DIN EN ISO 8692:2012-06 3.2effective concentrationconcentration of the test sample
43、 (ECx) at which an effect of x % is measured, if compared to the controlNOTE To unambiguously denote an EC value deriving from growth rate, it is proposed to use the symbol “ErC”.3.3lowest ineffective dilutionLIDdilution level at which no inhibition, or only effects not exceeding the test-specific v
44、ariability, are observedNOTE Adapted from ISO 15088:200713, 3.5.3.4specific growth ratemproportional rate of increase in cell density per time: =1nntddwheren is the cell density, expressed in cells per millilitre;t is the time, expressed in days.NOTE Specific growth rate is expressed in reciprocal d
45、ays (day1).ISO/TR 11044:2008, 3.24 PrincipleMonospecies algal strains are cultured for several generations in a defined medium containing a range of concentrations of the test sample, prepared by mixing appropriate quantities of growth medium, test sample, and an inoculum of exponentially growing al
46、gal cells. The test batches are incubated for a period of (72 2) h during which the cell density in each test solution is measured at least every 24 h.Inhibition is measured as a reduction in specific growth rate relative to control cultures grown under identical conditions.5 Reagents and media5.1 T
47、est organism, using either of the following planktonic fresh water algae species:a) Desmodesmus subspicatus (R. Chodat) E. Hegewald et A. Schmidt1)(86.81 SAG2);b) Pseudokirchneriella subcapitata (Korshikov) Hindak3)(ATCC 22662TM,2)CCAP 278/42)or 61.81 SAG2).NOTE 1 The two species do not show identic
48、al responses to toxic agents.NOTE 2 Both algae species are planktonic green algae belonging to the order of Sphaeropleales (Chlorophyta, Chlorophyceae) and are usually unicellular in culture.1) This species is formerly known as Scenedesmus subspicatus Chodat.2) Trade names of strains are examples of suitable strains available commercially. This information is given for the convenience of users of this document and does not constitute an endorsement by ISO of these products.3) This specie
