1、 STDBBSI BS EN IS0 73LIb-3-ENGL 1998 D Lb2LIbbS Ob735 BRITISH STANDARD Water quality - Determination of the acute lethal toxicity of substances to a fresh water fish Bruchgduno rerio Hamilton-Buchanan (Teleostei, Cyprinidae) - Flow-through method 1 The European Standard EN IS0 7346-3 : 1997 has the
2、status of a British Standard ICs 13.060.01 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW OL4 = BS EN IS0 BS 6068 : geetion 5.4 : 1998 7346-3 : 1998 BS EN IS0 73463 : 1998 National foreword This British Standard is the English language version of EN IS0 7346-3 : 1997. It is i
3、dentical with IS0 7346-3 : 1996. It supersedes BS 6068 : Section 5.4 : 1985 which is withdrawn. The UK participation in its preparation was entrusted by Technical Committee EW3, Water quality, to Subcommittee EW3/5, Biological methods, which has the responsibility to: - aid enquirers to understand t
4、he text; - present to the responsible internationalEuropean 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
5、subcommittee can be obtained on request to its secrew. BS EN IS0 73463 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 BS 6068 : Section 5.4. The various Section
6、s of BS 6068 are comprised within Parts 1 to 7, which, together with Part O, are listed below. Part O Introduction Part 1 Glossaq Part 2 Physica, chemical and biochemical methods Part 3 MWlQgiCd methods Part 4 M.irrobidogical methods Part 5 Biological methods Part 6 Sampling Part 7 Precision and acc
7、umcy Cross-references Attention 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 publicati
8、ons may be found in the BSI Standards Catalogue under the section entitled International Standards Correspondence Index, or by using the Find facility of the BSI Standards Electronic Catalogue. Compliance with a British Standard does not of itself confer immunity from legai obligations. Summary of p
9、ages This document comprises a front cover, an inside front cover, the EN IS0 title page, the EN IS0 foreword page, the Is0 title page, pages ii and iii, a blank page, pages 1 to 11, a blank page, an inside back cover and a back cover. This British Standard, having been prepared under the direction
10、of the Heaith and Environment Sector Board, was published under the authority of the Standards Board and comes into effect on 15 January 1998 Amendments issued since publication AmdNo. I Date I Text affected O BSI 1998 ISBN O 680 28490 5 STD-BSI BS EN IS0 734b-3-ENGL 1778 lb24bb7 Ob73531 772 EUROPEA
11、N STANDARD NORME EUROPENNE EUROPISCHE NORM EN IS0 73464 November 1997 ICs 13.060.01 Descriptors: see IS0 document English version Water quality - Determination of the acute lethal toxicity of substances to a freshwater fish Brachydanio rerio Hamilton- Buchanan (Teleostei, Cyprinidae) - Part 3: Flow-
12、through method (IS0 7346-3:1996) Qualit de leau - Dennination de la toxicit aigu8 Male de subtances vis-vis dun poisson deau douce I 8ra-b renb Hamiilon-Buchanan (Tloslei, Cyprinidae) I - Partie 3: Mthode avec renouvellement continu (IS0 7346-3:1996) Wasseheschaffenheil - Bestimmung der akuten letal
13、en Toxiritt von Substanzen gegenber einem SwasseAech Bmchycbb renb Hamillon-Buchanan (Teieostei, Cyprinidae) -Teil 3: Durchuverfahren (IC0 7346-3:1996) This European Standard was approved by CEN on 30 October 1997. CEN members are bound to comply with the CEWCENELEC Inlemai Regulations which slipdat
14、e lhe conditions for giving this European Slandard the 8tatus of a national standard without any alteration. Up-to-date iists and bibliographical references concerning such national standards may be obtained on application to lhe Central Secretariat or to any CEN member. This European Slandard exist
15、s in three offil versions (English, French, Qerman). A version in any other language made by translation under the responsibility of a CEN member into Is own language and notified to the Central Secretariat has the same status as the official versions. CEN members are the national standards bodies o
16、f Austria, Belgium, Czech Repubiic, Denmark, Finland, France, Oermany, Qreece, Icelend, Ireland, llaly, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMlTIEE FOR STANDARDIZATION COMITE EUROPEEN DE NORMALISATION EUROPISCHES KOMITEE FOR NORMUNG Ce
17、ntral Secretariat rue dc Staasart. 36 8-1050 Biuuel8 O 1997 CEN All rights of exploitation in any form and by any means reserved woridwide for CEN national Members. Ref. No. EN IS0 7346-31997 E STD-BSI BS EN IS0 73qb-3-ENGL 1998 Lb24bb9 b73532 b09 EN IS0 7346-3 : 1997 Foreword The text of the Intern
18、ational Standard from Technical Committee ISO/TC 147 “Water quality“ of the International Organization for Standardization (IS01 has been taken over as an European Standard by Technical Cornmittee CEN/TC 230 “Water analysis“, the secretariat of which is held by DIN. This European Standard shall be g
19、iven the status of a national standard, either by publication of an identical text or by endorsement, at the latest by May 1998, and conflicting national standards shall be withdrawn at the latest by May 1998. According to the CENKENELEC Internal Regulations, the national standards organizations of
20、the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of th
21、e International Standard IS0 7346-3:1996 has been approved by CEN as a European Standard without any modification. STD.BSI BS EN IS0 734b-3-ENGL 1798 Lb24bb7 Ob73533 545 EN IS0 7346-3 : 1997 I N T ER NAT I O NA L STANDARD IS0 734613 Second edition 1996-06-1 5 Water quality - Determination of the acu
22、te lethal toxicity of substances to a freshwater fish Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) - Part 3: Flow-through method * 2 Qualit de leau - Dtermination de la toxicit aigu ltale de substances vis-vis dun poisson deau douce Brachydanio rerio Hamilton-Buchanan (Tlostei, Cyprin
23、idae0 - Partie 3: Mthode avec renouvellement continu Reference number IS0 7346-3:1996(E) _ STD.BSI BS EN IS0 734b-3-ENGL 1998 m Lb24bb7 b73534 481 m EN IS0 7346-3 : 1997 Foreword IS0 (the International Organization for Standardization) is a worldwide federation of national standards bodies (IS0 memb
24、er bodies). The work of preparing International Standards is normally carried out through IS0 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. International organizations, government
25、al and non-governmental, in liaison with ISO, also take part in the work. IS0 collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. Draft International Standards adopted by the technical committees are circulated to the memb
26、er bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. International Standard IS0 7346-3 was prepared by Technical Committee iSO/TC 147, Water quality, Subcommittee SC 5, Biological methods. This second edition cancels a
27、nd replaces the first edition (IS0 7346-3:1984), which has been technicaliy revised. IS0 7346 consists of the following parts, under the general title Water quality - Determination of the acute lethal toxicity of substances to a freshwater fish Brachydanio rerio Hamilton-Buchanan fleleostei, Cyprini
28、daefl : - Part 1: Static method - Part 2: Semi-static method - Part 3: Flow-through method Annexes A, B and C of this part of IS0 7346 are for information only. D.rciiptor8: water, quality, water pollution, tests, water tests, biological tests, determination, toxicity. ii EN IS0 7346-3 : 1997 Introd
29、uction The three parts of IS0 7346 describe methods of determining the acute lethal toxicity of substances to the zebra fish (Brachydanio rerio Hamilton-Buchanan) but it must be emphasized that the recommended use of the zebra fish does not preclude the use of other species. The methodologies presen
30、ted here may also be used for other species of freshwater, marine or brackish water fish, with appropriate modifications of, for example, dilution water quality and the temperature conditions of the test. Within the three parts of IS0 7346, a choice can be made between static, semi-static and flow-t
31、hrough methods. The static test, described in IS0 7346-1, in which the solution is not renewed, has the advantage of requiring simple apparatus, 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
32、-through method, described in this part of IS0 7346, in which the test solution is replenished continuously, overcomes such problems but requires the use of more complex appar- atus. In the semi-static procedure, described in IS0 7346-2, the test sol- utions are renewed every 24 h or 48 h, this meth
33、od being a compromise between the other two. The flow-through method can be used for most types of substances, in- cluding those unstable in water, but the concentrations of the test sub- stance are determined wherever possible. The static method is limited to the study of substances whose tested co
34、ncentrations remain relatively constant during the test period. The semi-static method can be used for testing those substances whose concentrations can be maintained satis- factorily throughout the test by renewal of the solutions every 24 h or 48 h. Special arrangements may be necessary for substa
35、nces which are highly volatile. To assist in the preparation and maintenance of concentrations of sub- stances which may be lethal at concentrations close to that of their aqueous solubility, a small volume of solvent may be used, as specified in the methods. . li1 - STD.BSI BS EN IS0 734b-3-ENGL 19
36、98 Lb24bb9 Ob7353b 254 D EN IS0 7346-3 : 1997 Water quality - Determination of the acute lethal toxicity of substances to a freshwater fish Brachyanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) - Part 3: Flow-through method 1 Scope This part of IS0 7346 specifies a flow-through method for the d
37、etermination of the acute lethal toxicity of stable, non-volatile, single substances, soluble in water under specified conditions, to a freshwater fish Brachyanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) - common name, zebra fish in water of a specified quality. % % ;Il The method is applicab
38、le for assigning, for each test substance, broad categories of acute lethal toxicity to Brachyanio rerio under the test conditions. The results are insufficient by themselves to define water quality standards for environmental protection. The method is also applicable when using certain other specie
39、s of freshwater fish as the test organism”. The method may be adapted for use with other freshwater fish and marine and brackish water fish with appropriate modification of the test conditions, particularly with respect to the quantity and quality of the dilution water and the temperature. 2 Princip
40、le Determination, under specif ied conditions, of the concentrations at which a substance is lethal to 50 % of a test population of Brachyanio rerio after exposure periods of 24 h, 48 h, 72 h and 96 h to that substance in the ambient water. These median lethal concentrations are designated the 24 h
41、- LC50, 48 h - LC50. 72 h - LC50 and 96 h - LC50. The test is carried out in two stages: a) a preliminary test which gives an approximate in- dication of the acute median lethal concentrations and serves to determine the range of concen- trations for the final test; b) a final test, the results of w
42、hich alone are re- ported. 1) The following species of freshwater fish can be used, in addition to Brachydanio rerio, without modification to this part of IS0 7346. - Lepomis macrochirus (Teleostei, Centrarchidae - Oryzias latipes (Teleostei, Poeciliidae) - Pimephales promelas (Teleostei, Cyprinidae
43、) - Poecilia rericulara (Teleostei, Poeciliidae) Previous page is blank 1 STD.BSI BS EN IS0 73qb-3-ENGL 1998 m LbZLibb Ob73537 290 EN IS0 7346-3 : 1997 Where evidence is available to show that test con- centrations remain relatively constant (.e. within about 20 % of the nominal values) throughout t
44、he 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 about 80 %, or greater than 120 %, of the nominal values, then the analytical values are used in estim
45、ating 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 chemi- cal have declined significantly at any stage during the test, then, irrespec
46、tive of whether or not chemical analytical data are available, the LC50 cannot be de- fined 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 estimated from the nominal concentration
47、s used. 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 approximately 250 mg/l, expressed as calcium carbonate, and shall contain the following concen- trations of salts dissolved in distilled or deionized water: 294,O
48、mg/l CaC1,.2H20 123,3 mg/l MgSO4-7H,O 63,O mg/l NaHCO, 5,5 mg/l KCI Aerate the dilution water until the concentration of dissolved oxygen reaches at least 90 % of its air saturation value (ASV) and the pH is constant at 7.8 f 0,2. If necessary, adjust the pH of the solution by adding sodium hydroxid
49、e solution or hydrochloric acid. The dilution water thus prepared shall receive no further forced aeration before use in the tests. 3 Test organism and reagents 3.3 Stock solutions of test substances The reagents shall be of recognized analytical grade. The water used for the preparation of solutions shall be glassdistilled water or deionized water of at least equivalent purity. 3.1 Test organism The test species shall be Brachydanio rerio Hamilton- Buchanan (Teleostei, Cyprinidae), commo
