1、BRITISH STANDARD BS EN ISO 7346-1:1998 BS 6068-5.2: 1998 Water quality Determination of the acute lethal toxicity of substances to a fresh water fish Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) Static method The European Standard EN ISO 7346-1:1997 has the status of a British Standar
2、d ICS 13.060.01BSEN ISO 7346-1:1998 This British Standard, having been prepared under the directionof the Health and Environment Sector Board, waspublished under the authorityof the Standards Boardand comes into effect on 15January 1998 BSI 03-1999 ISBN 0 580 28488 3 Amendments issued since publicat
3、ion Amd. No. Date CommentsBSEN ISO 7346-1:1998 BSI 03-1999 i Contents Page National foreword ii Foreword 2 Text of EN ISO 7346-1 1BSEN ISO 7346-1:1998 ii BSI 03-1999 National foreword This British Standard is the English language version of EN ISO 7346-1:1997. It is identical with ISO 7346-1:1996. I
4、t supersedes BS 6068-5.2:1985 which is withdrawn. The UK participation in its preparation was entrusted by Technical Committee EH/3, Water quality, to Subcommittee EH/3/5, Biological methods, which has the responsibility to: aid enquirers to understand the text; present to the responsible internatio
5、nal/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. A list of organizations represented on this subcommittee can be obtained on request to its s
6、ecretary. BS EN ISO 7346-1 is one of a series of standards on water quality, others of which have been, or will be, published as Sections of BS 6068. This standard has therefore been given the secondary identifier BS6068-5.2. The various Sections of BS 6068 are comprised within Parts 1 to 7, which,
7、together with Part 0, are listed below. Part 0: Introduction; Part 1: Glossary; Part 2: Physical, chemical and biochemical methods; Part 3: Radiological methods; Part 4: Microbiological methods; Part 5: Biological methods; Part 6: Sampling; Part 7: Precision and accuracy. Cross-references Attention
8、is drawn to the fact that CEN and CENELEC Standards normally include an annex which lists normative references to international publications with their corresponding European publications. The British Standards which implement these international or European publications may be found in the BSI Stan
9、dards Catalogue under the section entitled “International Standards Correspondence 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
10、for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, pages i and ii, theEN title page, page 2, the ISO title page, pages ii and iii, pages 1 to 8, an
11、inside back cover and a back cover. This standard has been updated (see copyright date) and may have had amendments incorporated. This will be indicated in the amendment table on theinside front cover.EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 7346-1 November 1997 ICS 13.060.01 Descrip
12、tors: See ISO document English version Water quality Determination of the acute lethal toxicity of substances to a freshwater fish Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) Part 1: Static method (ISO 7346-1:1996) Qualit de leau Dtermination de la toxicit aigu ltale de substances vi
13、s-vis dun poisson deau douce Brachydanio rerio Hamilton-Buchanan (Tlostei, Cyprinidae) Partie 1: Mthode statique (ISO 7346-1:1996) Wasserbeschaffenheit Bestimmung der akuten letalen Toxizitt von Substanzen gegenber einem Swasserfisch Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) Teil 1
14、: Statisches Verfahren (ISO 7346-1:1996) This European Standard was approved by CEN on 30 October 1997. 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.
15、 Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the Central Secretariat 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
16、under the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy,
17、Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom. CEN European Committee for Standardization Comit Europen de Normalisation Europisches Komitee fr Normung Central Secretariat: rue de Stassart, 36 B-1050 Brussels 1997 CEN All rights of exploitation in any form
18、and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 7346-1:1997 EENISO 7346-1:1997 BSI 03-1999 2 Foreword The text of the International Standard from Technical Committee ISO/TC 147 “Water quality” of the International Organization for Standardization (ISO) has been taken ov
19、er as an European Standard by 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 of an identical text or by endorsement, at the latest by May 1998, and conflicting na
20、tional standards shall be withdrawn at the latest by May 1998. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece
21、, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of the International Standard ISO 7346-1:1996 has been approved by CEN as a European Standard without any modification.ENISO 7346-1:1997 ii BSI 03-1999
22、 Contents Page Foreword ii Introduction 1 1 Scope 1 2 Principle 1 3 Test organism and reagents 2 4 Apparatus 2 5 Test environment 3 6 Procedure 3 7 Expression of results 4 8 Test report 5 Annex A (informative) Environmental parameters for maintenance and breeding of zebra fish (Brachydanio rario Ham
23、ilton-Buchanan) 6 Annex B (informative) Suggested form for recording data 7 Annex C (informative) Bibliography Inside back cover Figure 1 Graphical interpolation of LC50 (linear scales) 4 Figure 2 Graphical interpolation of LC50 (logarithmic and probability scales) 5EN ISO7346-1:1997 BSI 03-1999 iii
24、 Foreword ISO (the International Organization for Standardization) is a worldwide 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
25、technical committee has been established has the right to be represented on that 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 matter
26、s of electrotechnical standardization. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. International Standard ISO 7346-1
27、was prepared by Technical Committee ISO/TC147, Water quality, Subcommittee SC5, Biological methods. This second edition cancels and replaces the first edition (ISO7346-1:1984), which has been technically revised. ISO 7346 consists of the following parts, under the general title Water quality Determi
28、nation of the acute lethal toxicity of substances to a freshwater fish Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae): Part 1: Static method; Part 2: Semi-static method; Part 3: Flow-through method. Annex A, Annex B and Annex C of this part of ISO 7346 are for information only. Descript
29、ors: Water, quality, water pollution, tests, water tests, biological tests, determination, toxicity.iv blankENISO 7346-1:1997 BSI 03-1999 1 Introduction The three parts of ISO 7346 describe methods of determining the acute lethal toxicity of substances to the zebra fish (Brachydanio rerio Hamilton-B
30、uchanan) but it must be emphasized that the recommended use of the zebra fish does not preclude the use of other species. The methodologies presented here may also be used for other species of freshwater, marine or brackish water fish, with appropriate modifications of, for example, dilution water q
31、uality and the temperature conditions of the test. Within the three parts of ISO 7346, a choice can be made between static, semi-static and flow-through methods. The static test, described in this part of ISO 7346, in which the solution is not renewed, has the advantage of requiring simple apparatus
32、, although the substances in the test vessel may become depleted during the course of the test and the general quality of the water may deteriorate. The flow-through method, described in ISO 7346-3, in which the test solution is replenished continuously, overcomes such problems but requires the use
33、of more complex apparatus. In the semi-static procedure, described in ISO 7346-2, the test solutions are renewed every 24 h or 48 h, this method being a compromise between the other two. The flow-through method can be used for most types of substances, including those unstable in water, but the conc
34、entrations of the test substance are determined wherever possible. The static method is limited to the study of substances whose tested concentrations remain relatively constant during the test period. The semi-static method can be used for testing those substances whose concentrations can be mainta
35、ined satisfactorily throughout the test by renewal of the solutions every 24h or 48h. Special arrangements may be necessary for substances which are highly volatile. To assist in the preparation and maintenance of concentrations of substances which may be lethal at concentrations close to that of th
36、eir aqueous solubility, a small volume of solvent may be used, as specified in the methods. 1 Scope This part of ISO 7346 specifies a static method for the determination of the acute lethal toxicity of stable, non-volatile, single substances, soluble in water under specified conditions, to a freshwa
37、ter fish Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) common name, zebra fish in water of a specified quality. The method is applicable for assigning, for each test substance, broad categories of acute lethal toxicity to Brachydanio rerio under the test conditions. The results are ins
38、ufficient by themselves to define water quality standards for environmental protection. The method is also applicable when using certain other species of freshwater fish as the test organism 1) . The method may be adapted for use with other freshwater fish and marine and brackish water fish with app
39、ropriate modification of the test conditions, particularly with respect to the quantity and quality of the dilution water and the temperature. 2 Principle Determination, under specified conditions, of the concentrations at which a substance is lethal to50% of a test population of Brachydanio rerio a
40、fter exposure periods of24h, 48h, 72h and 96h to that substance in the ambient water. These median lethal concentrations are designated the 24h LC50, 48h LC50, 72h LC50 and96h LC50. The test is carried out in two stages: a) a preliminary test which gives an approximate indication of the acute median
41、 lethal concentrations and serves to determine the range of concentrations for the final test; b) a final test, the results of which alone are reported. 1) The following species of freshwater fish can be used, in addition to Brachydanio rerio, without modification to this part of ISO7346. Lepomis ma
42、crochirus (Teleostei, Centrarchidae) Oryzias latipes (Teleostei, Poeciliidae) Pimephales promelas (Teleostei, Cyprinidae) Poecilia reticulata (Teleostei, Poeciliidae)ENISO 7346-1:1997 2 BSI 03-1999 Where evidence is available to show that test concentrations remain relatively constant (i.e.within ab
43、out 20 % of the nominal values) throughout the test, then either measured or nominal concentrations are used in the estimation of the LC50. Where such analyses show that the concentrations present remain relatively constant but are less than about80%, or greater than 120%, of the nominal values, the
44、n the analytical values are used in estimating the LC50. Where evidence is not available to show that the test concentrations remained at an acceptable level throughout the test period, or where it is known (or suspected) that the concentrations of the test chemical have declined significantly at an
45、y stage during the test, then, irrespective of whether or not chemical analytical data are available, the LC50 cannot be defined using this test method. In these cases, the test is not necessarily invalidated but it can only be stated that the LC50 of the substance is # x mg/l, the value, x, being e
46、stimated from the nominal concentrations used. 3 Test organism and reagents The reagents shall be of recognized analytical grade. The water used for the preparation of solutions shall be glass-distilled water or deionized water of at least equivalent purity. 3.1 Test organism The test species shall
47、be Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae), commonly known as the zebra fish. Each test fish shall have a total length of 30mm 5mm, which, in principle, corresponds to a mass of 0,3g 0,1g. They shall be selected from a population of a single stock. This stock should have been acc
48、limatized and, in any case, maintained for at least 7d prior to the test in dilution water, continuously aerated using bubbled air (see 3.2), under conditions of water quality and illumination similar to those used in the test. They shall be fed as normal up to the 24 h period immediately preceding
49、the test. Test fish shall be free of overt disease or visible malformation. They shall not receive treatment for disease during the test or in the 2 weeks preceding the test. Subsequent to the test, fish remaining alive should be suitably disposed of. Environmental conditions for the maintenance and breeding of zebra fish are given inAnnex A. 3.2 Standard dilution water The freshly prepared standard dilution water shall have a pH of 7,8 0,2, and a calcium hardness of a
