[[4-[[2-(4-cyclohexylphenoxy)ethyl]ethylamino]-2-methylphenyl]methylene]malononitrile

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Justification for type of information:

Experimental study comparable to guideline study

Qualifier:

according to guideline

Guideline:

other: EU method C.1 (1992)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Principles of method if other than guideline:

A study was performed to assess the acute toxicity of test material to Danio rerio under static conditions.

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Reconstituted water prepared according to the recommendations of ISO 7346. This freshly prepared standard dilution water was used for the maintenance of the test animals under flow-through conditions and for the preparation of stock- and test solutions of the test item.

Details on test solutions:

To produce the only test concentration 5.4 mg of the test item were added to 5 litres of dilution water, then treated with an ultrasonic bath for 60 min. and afterwards stirred for 24 h on a magnetic stirrer. Finally undissolved particles of the test item were removed by filtration using a folded filter of pore size 7-12 μm.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Source : Aqua KloeGer (Germany)
Acclimatisation : Stock held since 2007-09-18 and acclimatised to the test conditions since then.
Feeding : Commercial fish food, daily. Discontinued 24 h prior to test.
Mortalities during acclimatisation period : < 5 %
Mean standard length (n = 14) : 3.3 cm (S.D. = 0.15 cm)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure.

Hardness:

11.1°dH (= 198 mg/L CaCO3).

Test temperature:

21.4 to 22.3°C

pH:

7.7

Dissolved oxygen:

8.4 mg/l

Nominal and measured concentrations:

1 mg/L

Details on test conditions:

Test vessels : glass aquaria holding 5 L of test media
Experimental design : 1 test concentration plus 1 control
7 animals per test concentration
no feeding during the exposure period
static system
Method of initiation : animals were placed in prepared media
Loading : 0.36 g body weight (wet weight) per litre
Photoperiod : 16 h light : 8 h dark
Photoperiod : 16 h light : 8 h dark
Temperature : 21.4 to 22.3°C
Aeration : gentle aeration via narrow glass tubes
Test item concentration/s : 1 mg/L
Method of administration : direct weighing
Medium renewal : none
Duration of exposure : 96 hours

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No toxic effect at against fishes at the limit of water solubility under exposure conditions.

Details on results:

The criterion of death used in this study was the absence of response to physical stimulation. In addition to observations on mortality at 2, 24, 48, 72 and 96 hours, type and incidence of sub-lethal effects compared with control fish were observed.

Sublethal observations / clinical signs:

Table 1:			Control
Abiotic parameters	0 h	24 h		48 h	72 h	96 h
Temperature [°C]	21.4	21.5		21.7	21.6	21.6
Oxygen [mg/L]	9.3	8.5		8.4	8.5	8.4
Oxygen [% saturation]	105	95		97	97	96
pH value	7.7	7.7		7.7	7.8	7.7
Mortality for zebra fish	2 h	24 h		48 h	72 h	96 h
(initial population: 7)
Absolute	0	0		0	0	0
Cumulative	0	0		0	0	0
Cumulative [%]	0	0		0	0	0
Chemical analysis	0 h	24 h		48 h	72 h	96 h
HPLC replicates [mg/L]	<0.002/	<0.002/		<0.002/	<0.002/	<0.002/
	<0.002	<0.002		<0.002	<0.002	<0.002
HPLC mean value [mg/L]	<0.002	<0.002		<0.002	<0.002	<0.002

Table 2:			1 mg/L
Abiotic parameters	0 h	24 h		48 h	72 h	96 h
Temperature [°C]	22.2	22.1		22.3	21.4	21.4
Oxygen [mg/L]	8.6	8.4		8.4	8.6	8.4
Oxygen [% saturation]	100	93		95	97	96
pH value	7.7	7.7		7.7	7.8	7.8
Mortality for zebra fish	2 h	24 h		48 h	72 h	96 h
(initial population: 7)
Absolute	0	0		0	0	0
Cumulative	0	0		0	0	0
Cumulative [%]	0	0		0	0	0
Chemical analysis	0 h	24 h		48 h	72 h	96 h
HPLC replicates [mg/L]	0.014	0.005		0.004	<0.002	<0.002
	0.005	0.007		0.003	<0.002	0.002
HPLC mean value [mg/L]	0.010	0.006		0.004	<0.002	<0.002

Validity criteria fulfilled:

not specified

Conclusions:

The median lethal effect concentration LC50 was observed to be >1 mg/l . No toxic effect at against fishes at the limit of water solubility under exposure conditions.

Executive summary:

A study was performed to assess the acute toxicity of test material to Danio rerio under static conditions. The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity in Directive 92/69/EEC (O.J. No. L383A, 1992) Part C, Method 1 'Acute toxicity for Fish' which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992).

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 1 mg/L of Macrolex Yellow 6G dissolved in water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and a folded filter. The water solubility of test material is very low i.e

< 0.0007 mg/L. Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. The median lethal effect concentration LC50 was observed to be >1 mg/l . No toxic effect at against fishes at the limit of water solubility under exposure conditions.

Description of key information

Short term toxicity to fish:

A study was performed to assess the acute toxicity of test material to Danio rerio under static conditions. The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity in Directive 92/69/EEC (O.J. No. L383A, 1992) Part C, Method 1 'Acute toxicity for Fish' which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992).

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 1 mg/L of Macrolex Yellow 6G dissolved in water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and a folded filter. The water solubility of test material is very low i.e

< 0.0007 mg/L. Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. The median lethal effect concentration LC50 was observed to be >1 mg/l . No toxic effect at against fishes at the limit of water solubility under exposure conditions. The water solubility of test material is < 0.0007 mg/L , hence indicating that aquatic toxicity is unlikely to occure

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1 mg/L

Additional information

Short term toxicity to fish:

A study was performed to assess the acute toxicity of test material to Danio rerio under static conditions. The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity in Directive 92/69/EEC (O.J. No. L383A, 1992) Part C, Method 1 'Acute toxicity for Fish' which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992).

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 1 mg/L of Macrolex Yellow 6G dissolved in water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and a folded filter. The water solubility of test material is very low i.e

< 0.0007 mg/L. Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. The median lethal effect concentration LC50 was observed to be >1 mg/l . No toxic effect at against fishes at the limit of water solubility under exposure conditions. The water solubility of test material is < 0.0007 mg/L , hence indicating that aquatic toxicity is unlikely to occure