SIST EN ISO 7346-1:1998
(Main)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)
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)
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 freshwater fish (Brachydanio rerio - zebra fish). Replaces the first edition.
Wasserbeschaffenheit - Bestimmung der akuten letalen Toxizität von Substanzen gegenüber einem Süßwasserfisch (Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) - Teil 1: Statisches Verfahren (ISO 7346-1:1996)
Dieser Teil der ISO 7346 legt ein statisches Verfahren zur Bestimmung der akuten letalen Konzentration von stabilen, nicht-flüchtigen, unter gegebenen Bedingungen wasserlöslichen Einzelsubstanzen gegenüber einem Süßwasserfisch (Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)) - Trivialnahme Zebrabärbling) in Wasser einer festgelegten Beschaffenheit fest.
Qualité de l'eau - Détermination de la toxicité aiguë létale de substances vis-a-vis d'un poisson d'eau douce (Brachydanio rerio Hamilton-Buchanan (Téléostei, Cyprinidae)( - Partie 1: Méthode statique (ISO 7346-1:1996)
La présente partie de l'ISO 7346 prescrit une méthode statique pour la détermination de la toxicité aiguë létale de substances simples, stables, non volatiles, solubles dans l'eau dans les conditions prescrites, vis-à-vis d'un poisson d'eau douce [Brachydanio rerio Hamilton-Buchanan (Téléostei, Cyprinidae) -- nom commun, poisson-zèbre], dans une eau de qualité donnée. La méthode est applicable pour chaque substance à expérimenter, pour donner une idée globale de la toxicité aiguë létale vis-à-vis du Brachydanio rerio, dans les conditions de l'essai. Les résultats sont en eux-mêmes insuffisants pour définir des normes de qualité de l'eau en vue de la protection de l'environnement. La méthode est également applicable à certaines autres espèces de poissons d'eau douce comme organismes pour essais1). La méthode peut être adaptée pour d'autres poissons d'eaux douce, marine ou saumâtre avec des modifications appropriées des conditions d'essai, concernant notamment la quantité et la qualité de l'eau de dilution et la température.
Kakovost vode - Ugotavljanje akutne smrtne strupenosti snovi s sladkovodnimi ribami (Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)) - 1. del: Statična metoda (ISO 7346-1:1996)
General Information
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Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN ISO 7346-1:1998
01-maj-1998
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SIST ISO 7346-1:1997
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ULEDPL %UDFK\GDQLRUHULR+DPLOWRQ%XFKDQDQ 7HOHRVWHL&\SULQLGDH GHO
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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)
Wasserbeschaffenheit - Bestimmung der akuten letalen Toxizität von Substanzen
gegenüber einem Süßwasserfisch (Brachydanio rerio Hamilton-Buchanan (Teleostei,
Cyprinidae) - Teil 1: Statisches Verfahren (ISO 7346-1:1996)
Qualité de l'eau - Détermination de la toxicité aiguë létale de substances vis-a-vis d'un
poisson d'eau douce (Brachydanio rerio Hamilton-Buchanan (Téléostei, Cyprinidae)( -
Partie 1: Méthode statique (ISO 7346-1:1996)
Ta slovenski standard je istoveten z: EN ISO 7346-1:1997
ICS:
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
SIST EN ISO 7346-1:1998 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN ISO 7346-1:1998
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SIST EN ISO 7346-1:1998
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SIST EN ISO 7346-1:1998
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SIST EN ISO 7346-1:1998
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SIST EN ISO 7346-1:1998
IS0 7346-l :1996(E)
Foreword
IS0 (the International Organization for Standardization) is a worldwide
federation of national standards bodies (IS0 member 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, governmental
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 member 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-l was prepared by Technical Committee
lSO/TC 147, Water quality, Subcommittee SC 5, Biological methods.
This second edition cancels and replaces the first edition
(IS0 7346-l : 1984), which has been technically 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 (Teleostei,
Cyprinidae) ‘J:
- 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.
0 IS0 1996
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced
or utilized in any form or by any means, electronic or mechanical, including photocopying and
microfilm, without permission in writing from the publisher.
International Organization for Standardization
Case Postale 56 l CH-1211 Geneve 20 l Switzerland
Printed in Switzerland
ii
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SIST EN ISO 7346-1:1998
0 IS0
IS0 7346-l :1996(E)
Introduction
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 presented 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-through methods. The static test, described in this
part of IS0 7346, 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-through method, described in
IS0 7346-3, in which the test solution is replenished continuously, over-
comes such problems but requires the use of more complex apparatus.
In the semi-static procedure, described in IS0 7346-2, the test solutions
are renewed every 24 h or 48 h, this method being a compromise be-
tween 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 concentrations 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 substances 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.
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SIST EN ISO 7346-1:1998
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SIST EN ISO 7346-1:1998
INTERNATIONAL STANDARD 0 ISO IS0 7346-l :1996(E)
Water quality - Determination of the acute lethal
toxicity of substances to a freshwater fish [B~&ydanio
rer;o Hamilton-Buchanan (Teleostei, Cyprinidae)] -
Part 1:
Static method
with appropriate modification of the test conditions,
1 Scope
particularly with respect to the quantity and quality of
the dilution water and the temperature.
This part of IS0 7346 specifies a static method for the
determination of the acute lethal toxicity of stable,
non-volatile, single substances, soluble in water under
2 Principle
specified conditions, to a freshwater fish
[ Brachydanio rerio Hamilton-Buchanan (Teleostei,
Determination, under specified conditions, of the
common name, zebra fish] in water of
Cyprinidae) -
concentrations at which a substance is lethal to
a specified quality.
50 % of a test population of Brachydanio rerio after
exposure periods of 24 h, 48 h, 72 h and 96 h to that
The method is applicable for assigning, for each test
substance in the ambient water. These median lethal
substance, broad categories of acute lethal toxicity to
concentrations are designated the 24 h - LC50,
Brachydanio rerio under the test conditions.
48 h - LC50, 72 h - LC50 and 96 h - LC50.
The results are insufficient by themselves to define
The test is carried out in two stages:
water quality standards for environmental protection.
a) a preliminary test which gives an approximate in-
The method is also applicable when using certain
dication of the acute median lethal concentrations
other species of freshwater fish as the test and serves to determine the range of concen-
OrganismI).
trations for the final test;
The method may be adapted for use with other b) a final test, the results of which alone are re-
freshwater fish and marine and brackish water fish 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)
- Poecilia reticulata (Teleostei, Poeciliidae)
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SIST EN ISO 7346-1:1998
IS0 7346=1:1996(E) 0 IS0
Where evidence is available to show that test con-
3.2 Standard dilution water
centrations remain relatively constant (i.e. within
about 20 % of the nominal values) throughout the
The freshly prepared standard dilution water shall
test, then either measured or nominal concentrations
have a pH of 7,8 + 0,2, and a calcium hardness of
-
are used in the estimation of the LC50. Where such
approximately 250 mg/l, expressed as calcium
analyses show that the concentrations present remain
carbonate, and shall contain the following concen-
relatively constant but are less than about 80 %, or
trations of salts dissolved in distilled or deionized
greater than 120 %, of the nominal values, then the
water:
analytical values are used in estimating the LC50.
Where evidence is not available to show that the test 294,0 mg/l CaCI,.2H,O
concentrations remained at an acceptable level
123,3 mg/l MgSOJH,O
throughout the test period, or where it is known (or
suspected) that the concentrations of the test chemi-
63,0 mg/l NaHCO,
cal have declined significantly at any stage during the
test, then, irrespective of whether or not chemical
5,5 mg/l KCI
analytical data are available, the LC50 cannot be de-
fined using this test method. In these cases, the test
Aerate the dilution water until the concentration of
is not necessarily invalidated but it can only be stated
dissolved oxygen reaches at least 90 % of its air
that the LC50 of the substance is < x mg/l, the value,
saturation value (ASV) and the pH is constant at
X, being estimated from the nominal concentrations
7,8 + 0,2. If necessary, adjust the pH of the solution
used.
by adding sodium hydroxide 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
The reagents shall be of recognized analytical grade.
3.3 Stock solutions of test substances
The water used for the preparation of solutions shall
be glass-distilled water or deionized water of at least
A stock solution of the test substance should be pre-
equivalent purity.
pared by dissolving a known amount of test sub-
stance in a defined volume of dilution water,
deionized water or glass-distilled water. To enable
stock solutions to be prepared and to assist in their
3.1 Test organism
transfer to the test vessels, substances of low
aqueous solubility may be dissolved or dispersed by
The test species shall be Brachydanio rerio Hamilton-
suitable means, including ultrasonic devices and or-
Buchanan (Teleostei, Cyprinidae), commonly known
ganic solvents of low toxicity to fish. If any such or-
as the zebra fish. Each test fish shall have a total
ganic solvent is used, its concentration in the test
length of 30 mm + 5 mm, which, in principle, corre-
-
solution shall not exceed 0,l ml/l, or the volume con-
sponds to a mass of 0,3 g + 0,l g. They shall be se-
taining 0,l g/l, whichever is the greater. Where a sol-
lected from a population of a single stock. This stock
vent is used, two sets of controls, one containing
should have been acclimatized and, in any case,
solvent at the maximum concentration used in any
maintained for at least 7 d prior to the test in dilution
test vessel and one without solvent or test substance,
water, continuously aerated using bubbled air (see
shall be included.
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
the test.
3.4 Test solutions
Test fish shall be free of overt disease or visible
Test solutions are prepared by adding apbropriate
malformation. They shall not receive treatment for
amounts of the stock solution of the test substance
disease during the test or in the 2 weeks preceding
to the dilution water to give the required concen-
the test. Subsequent to the test, fish remaining alive
trations. It is recommended that, when a stock sol-
should be suitably disposed of.
ution is prepared in distilled or deionized water, no
maintenance and more than 100 ml of stock solution should be added
Enviro nmental conditions for the
per 10 litres of dilution water.
breedi ng of ze bra fish are given in annex A.
2
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SIST EN ISO 7346-1:1998
0 IS0
IS0 7346=1:1996(E)
Maintain the temperature of the water in the stock
4 Apparatus
tanks at 23 “C + 1 “C (4.2).
All materials which may come into contact with any
liquid into which the fish are to be placed, or with
6.2 Limit test
which they may come into contact, shall be inert and
should not absorb the test substance significantly.
Using the procedures described in this part of
IS0 7346, a limit test may be performed at the limit
Usual laboratory equipment and the following.
of aqueous solubility under the conditions of the test
or at 100 mg/l, whichever is the lower, in order to
4.1 Test vessels, of sufficient capacity (which may
demonstrate that the 96 h - LC50 is greater than this
need to be greater than 10 litres), with a large area
concentration. If no fish die in the limit test, no further
of interface between the air and the test medium (of
testing is required.
about 800 cm* for 10 litres of medium) and equipped
with a securely fixed and close-fitting cover. The vol-
Perform the limit test using 10 fish, with the same
ume of the test vessels should be sufficient that a
number in the control(s).
loading rate of 1 g of fish per litre of water should not
NOTE 1 Binominal theory dictates that, when 10 fish are
be exceeded at any time during the test.
used, with zero mortality there is a 99,9 % confidence that
Before use, the test vessels shall be cleaned the 96 h - LC50 is greater than the limit-test concentration.
If mortalities occur, a complete study (see 6.3 and 6.4) may
thoroughly, for example with a non-ionic detergent
need to be considered. If sub-lethal effects are observed,
(followed by acid and solvent washes for substances
these should be recorded.
expected to adsorb strongly to the vessel).
4.2 Temperature control equipment, to regulate 6.3 Preliminary test
the temperature of the test solutions and the water
Add at least 2,5 litres, preferably 5 litres, of standard
in the stock tanks to 23 “C + 1 “C by a suitable
-
dilution water (3.2) to each of six test vessels (4.1)
method.
and aerate if necessary to restore the concentration
of dissolved oxygen to at least 90 % of its air satu-
4.3 Dip-net, made of nylon or of another chemically
ration value.
inert material, for the control vessels and another for
all the test vessels (4.1).
Prepare test solutions by adding appropriate amounts
of stock solution of the test substance (3.3) to five of
5 Test environment
the vessels in order to obtain an adequate geometric
range of concentrations, for example 1 000 mg/l;
The preparation and storage of solutions, the holding
100 mg/l; 10 mg/l; 1 mg/l and 0,l mg/l. Nothing is
of fish, and all the manipulations and tests shall be
added to the sixth vessel, which serves as a control.
carried out in premises with an atmosphere free from
The solutions shall be adjusted to and maintained at
harmful concentrations of airborne contaminants.
23 “C + 1 “C (4.2) and shall not be forcibly aerated
during the test.
Take care to avoid any unwanted disturbance that
may change the behaviour
...
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