2,2'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[N-(4-chloro-2,5-dimethoxyphenyl)-3-oxobutyramide]

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-08-27 - 2002-08-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The test was performed with GLP compliance following relevant guidelines. The results are plausible and well presented.

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Remarks:

Principles of Good Laboratory Practice (GLP) § 19a, German Chemical Law (ChemG), Annex 1 (May 8, 2001) (implementation of the OECD Principles of Good Laboratory Practice as published on March 23, 1999, Official Journal of the European Union L77/9

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

A nominal concentration of 100 mg/L was used for the preparation of a test solution at maximum saturation of the test substance. After treatment with ultrasonic waves for approx. 30 minutes the preparation was stirred for approximately 4 days to ensure that the solubility limit of the test substance in the test water is reached. The resulting dispersion was pre-filtered through a cellulose filter followed by filtration through a membrane filter of 0.45 µm pore size.
The tested concentration was visually present as clear solution.
Analytical monitoring of the concentration of the test substance in the test water was not conducted since no relevant results could be expected. The solubility limit of the test substance is below the analytical detection limit. From the properties of the test substance it could be concluded that the test substance is chemically stable in water over the test interval of 96 hours.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species : zebrafish, Danio rerio (HAMILTON-BUCHANAN)
Origin : Aventis Pharma Deutschland GmbH, ProTox
Date of hatching : February 28, 2002
Delivery date : May 02, 2002

The fish were kept for 14 days in water for dilution before the start of the study under the following conditions:
Temperature : 22 ± 1 °C
Oxygen content : >= 80 % of the saturation value
Duration of light period : 12 hours daily
Feeding : twice daily ad libitum
Food : Tetra Min, Tetra Werke, Meile (Germany)

The body length of 7 representative fish from each batch was measured:
Batch No.: 2/2002/L2 (31-Aug-2002); n=7; Body length (Variation range): 2.5 to 2.7; (cm) mean: 2.6; s: ± 0.08

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Hardness:

The total hardness of the dilution water was determined weekly and was at 2.7 mmol Ca^2+ + Mg^2+/L during the study.

Test temperature:

Temperature (°C)
Concentration group: 21.2 to 21.3
Control group: 21.2 to 21.3

pH:

Concentration group: 7.5 to 7.9
Control group: 7.3 to 7.9

Dissolved oxygen:

Oxygen content (mg/L):
Concentration group: 6.5 to 8.6
Control group: 7.0 to 9.0

Salinity:

Water quality was according to ISO/DIS 73461/1

Nominal and measured concentrations:

A nominal concentration of 100 mg/L was used for the preparation of a test solution at maximum saturation of the test substance, see above.

Details on test conditions:

The study was conducted in a static system. The test chambers, which were calibrated to 4 liters, were made of glass (length 20 cm, width 15 cm, height 20 cm) and stood in a water bath made of Hostalit Z® with a Plexiglas viewing panel. The temperature of the water bath was regulated by a thermostat to 22 ± 1 °C. The chambers were illuminated from above from 06.00 a.m. to 06.00 p.m. The light intensity directly over the chambers was approximately 700 lux.
The test chambers were not aerated during the course of the study.
After the stirring period and the filtration the test preparation was aerated before Insertion of the test fish using glass capillaries with a bubble-frequency of 1 - 3 per second.

Study groups
There were 7 fish in each group (control and nominal test concentration of 100 mg/L).

Test procedure
After the test concentration had been prepared and water parameters recorded, 7 fish were assigned to each test and control chamber. The fish were not fed during the entire study period. Inspection of the fish took place after 3, 6, 24, 48, 72 and 96 hours and involved recording the mortality and visible alterations in appearance and behavior. The water parameters were measured and recorded before study start and after 0, 24, 48, 72 and 96 hours.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Observations
The 100 mg/L group showed no effect in comparison with the control. In the control group no effects were observed.

Mortality
In this 96-hour acute toxicity study of the submission substance in zebrafish (Danio rerio) mortality occurred neither in the 100 mg/L-group (nominal concentration) nor in the control group

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

not applicable

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this test the LC50 of the submission substance after 96 hours was > 100 mg/L (based an nominal concentration) and is above the solubility limit of the test substance in the test water.

Executive summary:

The submission substrance was tested in zebrafish (Danio rerio) for 96 hours in a static system.

The test was performed with GLP compliance and following relevant guidelines with reliability category 1.

The nominal concentration tested was 100 mg/L and a control (0 mg/L).

A nominal concentration of 100 mg/L was used for the preparation of a test solution at maximum saturation of the test substance. After treatment with ultrasonic waves for approx. 30 minutes the preparation was stirred for approximately 4 days to ensure that the solubility limit of the test substance in the test water was reached. The resulting dispersion was pre-filtered through a cellulose filter followed by filtration through a membrane filter of 0.45 µm pore size.

The tested concentration was visually present as clear solution. Analytical monitoring of the concentration of the test substance in the test water was not conducted since no relevant results could be expected. The solubility limit of the test substance was below the analytical detection limit. From the properties of the test substance it could be concluded that the test substance is chemically stable in water over the test interval of 96 hours.

In this 96-hour acute toxicity study the 100 mg/L group showed no effect in comparison to the control. In the control group no effects were observed.

Under the conditions of this test the LC₅₀ of the submission substance after 96 hours was > 100 mg/L (based on nominal concentration) and is far above the solubility limit of the test substance in the test water.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2002-08-27 - 2002-08-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The test was performed with GLP compliance following relevant guidelines. The results are plausible and well presented.

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Remarks:

Principles of Good Laboratory Practice (GLP) § 19a, German Chemical Act (ChemG), Annex 1 (May 8, 2001)

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

A nominal concentration of 100 mg/L was used for the preparation of a test solution at maximum saturation of the test substance. After treatment with ultrasonic waves for approx. 30 minutes the preparation was stirred for approximately 4 days to ensure that the solubility limit of the test substance in the test water is reached. The resulting dispersion was pre-filtered through a cellulose filter followed by filtration through a membrane filter of 0.45 µm pore size.
The tested concentration was visually present as clear solution.
Analytical monitoring of the concentration of the test substance in the test water was not conducted since no relevant results could be expected. The solubility limit of the test substance is below the analytical detection limit. From the properties of the test substance it could be concluded that the test substance is chemically stable in water over the test interval of 96 hours.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species : zebrafish, Danio rerio (HAMILTON-BUCHANAN)
Origin : Aventis Pharma Deutschland GmbH, ProTox
Date of hatching : February 28, 2002
Delivery date : May 02, 2002

The fish were kept for 14 days in water for dilution before the start of the study under the following conditions:
Temperature : 22 ± 1 °C
Oxygen content : >= 80 % of the saturation value
Duration of light period : 12 hours daily
Feeding : twice daily ad libitum
Food : Tetra Min, Tetra Werke, Meile (Germany)

The body length of 7 representative fish from each batch was measured:
Batch No.: 2/2002/L2 (31-Aug-2002); n=7; Body length (Variation range): 2.5 to 2.7; (cm) mean: 2.6; s: ± 0.08

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Hardness:

The total hardness of the dilution water was determined weekly and was at 2.7 mmol Ca2+ + Mg2+/L during the study.

Test temperature:

Temperature (°C)
Concentration group: 21.2 to 21.3
Control group: 21.2 to 21.3

pH:

Concentration group: 7.4 to 7.9
Control group: 7.3 to 7.9

Dissolved oxygen:

Oxygen content (mg/L):
Concentration group:6.3 to 8.5
Control group: 7.0 to 9.0

Salinity:

Water quality was according to ISO/DIS 73461/1

Nominal and measured concentrations:

A nominal concentration of 100 mg/L was used for the preparation of a test solution at maximum saturation of the test substance, see above.

Details on test conditions:

The study was conducted in a static system. The test chambers, which were calibrated to 4 liters, were made of glass (length 20 cm, width 15 cm, height 20 cm) and stood in a water bath made of Hostalit Z® with a Plexiglas viewing panel. The temperature of the water bath was regulated by a thermostat to 22 ± 1 °C. The chambers were illuminated from above from 06.00 a.m. to 06.00 p.m. The light intensity directly over the chambers was approximately 700 lux.
The test chambers were not aerated during the course of the study.
After the stirring period and the filtration the test preparation was aerated before Insertion of the test fish using glass capillaries with a bubble-frequency of 1 - 3 per second.

Study groups
There were 7 fish in each group (control and nominal test concentration of 100 mg/L).

Test procedure
After the test concentration had been prepared and water parameters recorded, 7 fish were assigned to each test and control chamber. The fish were not fed during the entire study period. Inspection of the fish took place after 3, 6, 24, 48, 72 and 96 hours and involved recording the mortality and visible alterations in appearance and behavior. The water parameters were measured and recorded before study start and after 0, 24, 48, 72 and 96 hours.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Observations
The 100 mg/L group showed no effect in comparison with the control. In the control group no effects were observed.

Mortality
In this 96-hour acute toxicity study of the test substance in zebrafish (Danio rerio) mortality occurred neither in the 100 mg/L-group (nominal concentration) nor in the control group

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

not applicable

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this test the LC50 of the submission substance after 96 hours was > 100 mg/L (based an nominal concentration) and is above the solubility limit of the test substance in the test water.

Executive summary:

The submission substance was tested in zebrafish (Danio rerio) for 96 hours in a static system.

The test was performed with GLP compliance and following relevant guidelines with reliability category 1.

The nominal concentration tested was 100 mg/L.

A nominal concentration of 100 mg/L was used for the preparation of a test solution at maximum saturation of the test substance. After treatment with ultrasonic waves for approx. 30 minutes the preparation was stirred for approximately 4 days to ensure that the solubility limit of the test substance in the test water was reached. The resulting dispersion was pre-filtered through a cellulose filter followed by filtration through a membrane filter of 0.45 µm pore size.

The tested concentration was visually present as clear solution. Analytical monitoring of the concentration of the test substance in the test water was not conducted since no relevant results could be expected. The solubility limit of the test substance was below the analytical detection limit. From the properties of the test substance it could be concluded that the test substance is chemically stable in water over the test interval of 96 hours.

In this 96-hour acute toxicity study, the 100 mg/L group showed no effect in comparison to the control. In the control group no effects were observed.

Under the conditions of this test the LC₅₀ of the submission substance after 96 hours was > 100 mg/L (based on nominal concentration) and is far above the solubility limit of the test substance in the test water.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1994-01-31 - 1994-02-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Due to the poor solubility of the test item in solvent and water, no more than 0.1 mg/L had been tested in a limit test to avoid precipitation during the test.

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1400 (Fish Acute Toxicity Test)

Principles of method if other than guideline:

not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP-compliance in accordancve with directive 88/320 EEC; date of inspection

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
not applicable

Analytical monitoring:

yes

Details on sampling:

Verification of Test Concentrations
Chemical analysis of the test preparations at 0, 24 and 96 hours showed the measured test concentrations to be near nominal. It was therefore considered justifiable to estimate the LC50 values in terms of nominal test concentrations only.

The test material concentration in the test samples was determined spectrophotometrically.

Samples
An aliquot (500 ml) of each test sample was extracted with three portions (3 x 50 ml) of dichloromethane, each extract being filtered through anhydrous sodium sulphate. The combined extracts were evaporated to dryness and the residue dissolved in dichloromethane such that the final theoretical test concentration was a nominal 2.5 mg/l.

Stability:
Pre-study test samples were prepared and analysed initially and then after storage at ambient temperature in light and dark conditions over a 24-hour period (equivalent to period of media renewal). The results obtained were:
114% of nominal initially,
94.7% of nominal after 24h at ambient temperature in light,
97.4% of nominal after 24h at ambient temperature in the dark.
The test samples have been considered to be sufficiently stable for the purposes of this study.

Vehicle:

yes

Details on test solutions:

Preliminary solution in dimethylformamide.
200 mg of test material was dispersed in solvent and the volume adjusted to 100 mL to give a 200 mg/100 mL solvent stock dispersion. 1 mL of this solvent stock solution was dispersed in 20 litres of dechlorinated tap water to give the test concentration 0.10 mg/L.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

Name Rainbow trout (Oncorhynchus mykiss)
Source Parkwood Trout Farm, Wigmore, Kent, UK.
Acclimatisation
Stock held since 4 January 1994 and acclimatised to test conditions from 24 - 31 January 1994.
The stock fish were maintained in a glass fibre tank with a "single pass" water renewal system. The lighting was controlled to give a 16 hours light and 8 hours darkness cycle.
Temperature 14.0°C
Dissolved Oxygen larger or equal to 9.1 mg O2/L
Feeding Commercial trout pellets daily. Discontinued 48h prior to test.
Mortalities 1% in the 7 days prior to the start of the test.
Medication None.
Mean standard length 4.6 cm (SD = 0.2 cm).
Mean weight 0.73 g (SD = 0.098 g).

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

not applicable

Hardness:

Total hardness 95 mg/L as CaCO3.

Test temperature:

14°C in the controls and the three test replicates.

pH:

controls: 7.4 - 7.6
test samples: 7.4 - 7.6

Dissolved oxygen:

[mg O2/L]
controls: 9.1 - 9.9
test samples: 9.1 - 9.9

Salinity:

not applicable

Nominal and measured concentrations:

Chemical analysis of the test preparations at 0, 24 and 96 hours showed the measured test concentrations to be near nominal. It was therefore considered justifiable to estimate the LC50 values in terms of nominal test concentrations only.

Details on test conditions:

Test Water
Laboratory tap water, dechlorinated by the addition of sodium thiosulphate. Total hardness 95 mg/L as CaCO3. Total Organic Carbon content 1.34 mg C/L and a particulate matter content of 0.8 mg/L.
Exposure Conditions
Test vessels : Glass aquaria holding 20 L of test media.
Experimental design :
1 test concentration in triplicate plus 1 control and 1 solvent control (including 50 µL/L auxiliary solvent) each in duplicate. 10 animals per replicate. This experimental design conforms to a "Limit Test" to confirm that at the highest attainable test concentration of 0.10 mg/L, no mortalities or adverse reactions to exposure were observed.

Method of initiation : Fish were placed, at random, in prepared test media.
Loading: 0.37 g bodyweight/litre.
Photoperiod: 16h light:8h dark.
Temperature: 14.0°C
Aeration: via narrow bore glass tubes.
Test concentration: 0.10 mg/L in triplicate. This test concentration was assigned following a preliminary range-finding study, where 3 fish were exposed to the following concentration for a period of 96 hours: 0.10 mg/L. The results showed no mortalities at the concentration of 0.10 mg/L. Preliminary solubility studies performed showed that at concentrations in excess of 0.10 mg/L a precipitate was formed on addition of the solvent stock dispersion to water. Therefore the range-finding study was only carried out at 0.10 mg/L.

Medium renewal: Daily. The study was performed using semi-static test conditions as opposed to dynamic, continuous flow test conditions due to the test material being observed to precipitate out of the solvent stock dispersion over a 24-hour period. However a good dispersion of the test material in solvent could be prepared from which a homogeneous aliquot could be sampled in order to prepare the test dispersions for a semi-static test regime.

Duration of exposure : 96 hours.

Environmental measurements : pH, dissolved oxygen concentration and temperature were recorded daily throughout the study.
Criteria of death: Absence of (i) respiratory movement and (ii) response to a physical stimulation.

Reference substance (positive control):

not required

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: zero mortalities and absence of any behavioural reactions to exposure

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: zero mortalities and absence of any behavioural reactions to exposure

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: zero mortalities and absence of any behavioural reactions to exposure

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: zero mortalities and absence of any behavioural reactions to exposure

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

There were no mortalities or other adverse reactions to exposure in 30 fish exposed to a concentration of 0.10 mg/L for a period of 96 hours.
Neither in the two controls, nor in the two solvent controls occurred any mortalities during the test period.

0.10 mg/L was the highest test concentration that could be prepared due to the limited solubility of the test material in water and auxiliary solvent and having due regard to the amount of auxiliary solvent permitted in the test under the EPA Guidelines. Other recognised auxiliary solvents were used during the preliminary range-finding experiments. However dimethylformamide was found to give the best testable dispersion of the test material in water. At higher test concentrations there was a marked precipitation of the test material on addition of the solvent stock solution to water.

Chemical analysis of the test preparations at 0, 24 and 96 hours showed the measured test concentrations to be near nominal. It was therefore considered justifiable to estimate the LC50 values in terms of nominal test concentrations only.

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

not applicable

Validity criteria fulfilled:

yes

Conclusions:

The test item had been tested on rainbow trout (Oncorhynchus mykiss) for acute toxicity (96h) in a limit test (0.1 mg/L due to poor solubility) according to OECD 203 and US EPA (title 40, part 797, section 1400) and compliant to GLP (reliability category 1). The NOEC for all time points (3, 6, 244, 48, 72, and 96 hours) had been determined as >= 0.1 mg/L and the LC50 (3, 6, 244, 48, 72, and 96 hours) as > 0.1 mg/L.

Executive summary:

The test item had been tested on rainbow trout (Oncorhynchus mykiss) for acute toxicity (96h) in a limit test according to OECD 203 and US EPA (title 40, part 797, section 1400) and compliant to GLP (reliability category 1). 0.10 mg/L was the highest test concentration that could be prepared due to the limited solubility of the test material in water and auxiliary solvent and having regard to the amount of auxiliary solvent permitted in the test under the EPA Guidelines. Neither in the controls and solvent controls, nor in the three test replicates of 10 fish, each occurred any mortalities or could behavioural abnormalities be observed. Therefore, the NOEC for all time points (3, 6, 244, 48, 72, and 96 hours) had been determined as

>= 0.1 mg/L and the LC50 (3, 6, 244, 48, 72, and 96 hours) as > 0.1 mg/L.

Description of key information

Under the conditions of this test the LC50 of the submission substance after 96 hours was > 100 mg/L (based an nominal concentration) and is above the solubility limit of the test substance in the test water.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Three acute toxicity studies of reliability 1 for Pigment Yellow 83, and its structural analogues, Pigment Yellow 12 and 170 on Danio rerio and Oncorhynchus mykiss are available. As could be expected from a grouping approach, for all three pigments of the Diarylide Yellow group consistently no toxicity towards fish has been found in any of these OECD 203 studies with nominal concentrations well above the solubility limit (0.4 μg/L).