N-[2-[(2-bromo-4,6-dinitrophenyl)azo]-5-(diallylamino)-4-methoxyphenyl]acetamide

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:

April 2-6, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Control mortality in one replicate was greater than 10%. However, since there was 10% mortality in the control fish of another test conducted at the same time using the same batch of stock, the overall health of the fish was considered satisfactory.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Stock of animals were maintained and the test was performed in dechlorinated mains water.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: Upwey Trout Farm, Weymouth, Dorset
- Length at study initiation (mean): 46.5 mm
- Weight at study initiation (mean): 0.67 g

ACCLIMATION
- Acclimation period: three weeks
- Acclimation conditions (same as test or not): not same as test, temperature - 7.9-17.6°C, dissolved oxygen - >80% air saturation
- Type and amount of food during acclimation: Trout Fry feed, B.P. Nutrition (UK) Limited, ad libitum

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

Test animals were observed for mortality at 24, 48, 72 and 96 h

Test temperature:

13-16.5°C

pH:

7.4-8.6

Dissolved oxygen:

76-100% air saturation

Nominal and measured concentrations:

Nominal: 0, 100, 180, 320, 560 and 1000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: plastic aquarium
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 10 L
- Aeration: yes, continuous diffusion of air
- No. of organisms per vessel: ten
- No. of vessels per concentration (replicates): two
- No. of vessels per control (replicates): two
- Biomass loading rate: not specified

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated mains water

OTHER TEST CONDITIONS
- Photoperiod: 16 h light: 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

189 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The results of this study indicate that the test substance was toxic to rainbow trout at a concentration of 100 mg/L and above. The highest concentration of 1000 mg/L resulted in 100% mortality at 48 h. The lowest test concentration which caused 100% mortality within 96 h was 560 mg/L. The highest no observed effect concentration could not be determined as 10% mortality was recorded in the lowest concentration of 100 mg/L.

The control mortality in the study was 20%.

Validity criteria fulfilled:

no

Conclusions:

Under the study conditions, the 96 h LC50 of the test substance to rainbow trout (Oncorhynchus mykiss) was determined to be 189 mg/L (nominal).

Executive summary:

A study was conducted to determine the toxicity of the test substance (100% purity) to rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203. The fish were exposed for 96 h to nominal concentrations of 0, 100, 180, 320, 560 and 1000 mg/L. Mortality and symptoms were assessed daily. No analytical verification of test concentrations was conducted. Exposure to 320 mg/L and above induced mortality from 24 h. At the lowest concentration there was 10% mortality at 96 h. Control mortality in one replicate was greater than 10%. However, since there was 10% mortality in the control fish of another test conducted at the same time using the same batch of stock, the overall health of the fish was considered satisfactory. Under the study conditions, the nominal 96 h LC50 was determined to be 189 mg/L (Seaber, 1994).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not specified

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Stock of animals were maintained and the test was performed in dechlorinated mains water.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: Upwey Trout Farm, Weymouth, Dorset
- Length at study initiation (mean): 51.8 mm
- Weight at study initiation (mean): 1.34 g

ACCLIMATION
- Acclimation period: 3 d
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: Trout Fry feed, B.P. Nutrition (UK) Limited, ad libitum

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

Test animals were observed for mortality at 24, 48, 72 and 96 h

Test temperature:

15.2-15.5°C

pH:

8.0-8.3

Dissolved oxygen:

86.9-93% air saturation

Nominal and measured concentrations:

Nominal: 0, 56, 100, 180, 320 and 560 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: plastic aquarium
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 10 L
- Aeration: yes, continuous diffusion of air
- No. of organisms per vessel: ten
- No. of vessels per concentration (replicates): two
- No. of vessels per control (replicates): two
- Biomass loading rate: not specified

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated mains water

OTHER TEST CONDITIONS
- Photoperiod: 16 h light: 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

91 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The results of this study indicate that the test substance was rapidly toxic to rainbow trout at a concentration of 560 mg/L. All of the exposed fish at this concentration died at 24 h. The lowest concentration of the sample which caused 100% mortality within 96 h was 180 mg/L. A no observed effect concentration could not be determined as 20% mortality was recorded in the lowest concentration of 56 mg/L within 96 h.

The control mortality in the study was within the required 10% level.

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the nominal 96 h LC50 of the test substance was determined to be 91 mg/L.

Executive summary:

A study was conducted to determine the toxicity of the test substance (43% purity) to rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203. The fish were exposed for 96 h to nominal concentrations of 0, 56, 100, 180, 320 and 560 mg/L (equivalent to 0, 20, 40, 72, 128 and 224 mg a.i./L). Mortality and symptoms were assessed daily. No analytical verification of test concentrations was conducted. Exposure to 320 mg/L and above induced mortality from 24 h. At the lowest concentration there was 20% mortality at 96 h. Under the study conditions, the nominal 96 h LC50 of the test substance was determined to be 91 mg/L (Butler, 1993).

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

other: raw data

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Raw data available, not GLP compliant

Principles of method if other than guideline:

Groups of 10 Zebra fish (Danio rerio) were exposed to the test substance for 96 h at nominal concentrations of 100, 300 and 1000 mg/L. Mortality was recorded at 1, 4, 24, 48, 72 and 96 h. Water temperature, pH and oxygen content were monitored.

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Number of animals: 10 zebra fish per test concentration
Age: 4-6 months
Size: about 3 cm
Weight: 0.25-0.35 g
Supplier: West Aquarium GmbH, D-3422 Bad Lauterberg

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

23°C

pH:

8 +/- 0.2

Dissolved oxygen:

7.8-8.3

Nominal and measured concentrations:

- Nominal: 100, 300 and 1000 mg test substance/L
- Active ingredient basis: 62.5, 187.5 and 625 mg/L

Details on test conditions:

- Water: 3 liters, water used according to ISO guidelines, in a full-glass basins, 5 liter basins (15x20x20 cm) tempered. Basins have been cleaned with RBS-35 solution (FLUKA 83462) and rinsed with clean water.
- Testing water should be kept at a pH between 6 and 8.5. In case of deviations, pH needs to be adjusted with NaOH or HCl solution.
- O2-concentration must be kept at 4.8 mg/L at a minimum. Sufficient oxygen supply must be ensured. O2-target concentration 6.4 mg/L
- Acclimatization period: minimum 12 days at 21-25 °C for Zebra fish.
- Feeding periods: daily (TETRA MIN, flocks, Dr. U.Baersch GmbH, D-4520 Melle, Germany)
- Day-night cycles: 12/12h
- Termination: For termination of the experiment MS-222 (Sandoz) should be used at 1% (Methacain).

Testing solution:
1.5 ml Stock-solution 1
1.5 ml Stock-solution 2
15 ml Stock-solution 3
add 1 liter of deionized water for zebrafish
where
stock solution 1 is 250 g CaCl x 2 H2O (Merck 2382) + 29 g NaCl (Merck 6404E) + 9 g NaNO3 (Merck 6537) add 1 liter of deionized water
stock solution 2 is 115 g MgSO x 7 H2O (Merck 5886) + 79 g Na2SO4 (Merck 822286) add 1 liter of deionized water
stock solution 3 is 27 g NaHO3 (Merck 6329E) + add 1 liter of deionized water

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 149 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

not specified

Conclusions:

Under the study conditions, the nominal 96 h LC50 of the test substance was determined to be ca. 149 mg/L.

Executive summary:

A study was conducted to determine the toxicity of the test substance (37.3% purity), based on total organic carbon content) to Zebra fish (Danio rerio). Ten fish/group were exposed to the test substance at nominal concentrations of 100, 300 and 1000 mg/L (equivalent to ca. 37, 112 and 373 mg a.i./L respectively). The animals were observed daily for mortality during 96 h. Oxygen, pH and temperature were monitored. No analytical validation of test concentrations was conducted. There was no mortality at 100 mg/L. All fish died within 48 h at 300 mg/L and within 4 h at 1000 mg/L. Under the study conditions, the nominal 96 h LC50 of the test substance was determined to be ca. 149 mg/L (Huntsman, 1986).

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: DIN 38412

Deviations:

not specified

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

no

Test organisms (species):

Leuciscus idus

Details on test organisms:

TEST ORGANISM
- Common name: Golden orfe
- Strain: Linnaeus
- Source: Fischzucht Paul Eggers, D-2354 Hohenwestedt, FRG
- Length at study initiation (mean and range): 8.4 cm (7.5-9.3)
- Weight at study initiation (mean and range): 6.0 g (4.4-7.6)

ACCLIMATION
- Acclimation period: 5 mo
- Acclimation conditions (same as test or not): same
- Type and amount of food during acclimation: Growing Feed F/B 50
- Feeding frequency during acclimation: ad libitum
- Health during acclimation (any mortality observed): No mortality

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

Mortality of the test animals were observd at 1, 4, 24, 48, 72 and 96 h during the exposure period

Hardness:

2.5 mmol/L

Test temperature:

20-21°C

pH:

7.0-7.4

Dissolved oxygen:

4.7-8.5 mg/L

Salinity:

The resalting was carried out by addition of: 294.0 mg/L CaCl2.2H2O, 123.3 mg/L MgSO4.7H2O, 63.0 mg/L NaHCO3, and 5.5 mg/L KCl

Conductivity:

max 10 micro mho

Nominal and measured concentrations:

0. 46.4, 100, 215, 464 and 1000 mg/L (nominal concentrations)

Details on test conditions:

TEST SYSTEM
- Test vessel: all glass aquarium
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 30 cm x 22 cm x 24 cm; 10 L
- Aeration: yes, continuous aeration with oil-free air
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): one
- No. of vessels per control (replicates): one
- Biomass loading rate: 6 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: fully demineralised tap water
- Acid capacity: 0.8 mmol/L
- Na/K ratio: 10:1
- Ca/mg ratio: 4:1
- Culture medium different from test medium: yes
- Intervals of water quality measurement: 1, 24, 48, 72 and 96 h

OTHER TEST CONDITIONS
- Adjustment of pH: not specified
- Photoperiod: 16 h light + 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.15

- Range finding study
- Test concentrations: not specified, LC50 between 100 and 1000 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 - < 215 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities:
- Observations on body length and weight: None
- Other biological observations: Apathy, tumbling and narcotic-like state at 1 and 4 h of exposure
- Any observations (e.g. precipitation): Undissolved test substance was visible on the bottom of the aquarium increasing with the increase in test concentrations

Validity criteria fulfilled:

not specified

Conclusions:

Under the study conditions, the nominal 96 h LC 50 of the test substance to golden orfe (Leuciscus idus L.) was determined to be >100<215 mg/L.

Executive summary:

A study was conducted to determine the toxicity of the test substance (in the form of aqueous dispersion of 40% purity) to golden orfe (Leuciscus idus L.) according to the guideline DIN 38412. The fish were exposed for 96 h to nominal concentrations of 0, 46.4, 100, 215, 464 and 1000 mg/L (equivalent to 0, 19, 40, 86, 186 and 400 mg a.i./L). Mortality and symptoms were assessed daily. No analytical verification of test concentrations was conducted. Apart from apathy, tumbling and narcotic-like state at 1 and 4 h, no symptoms were observed in any group. Exposure at 215, 464 and 1000 mg/L induced mortality from 4 h. Under the study conditions, the nominal 96 h LC 50 was determined to lie between 100 and 215 mg/L (Munk, 1990).

Description of key information

The structural analogue substance Disperse Blue 79:1 Br was used in four tests to determine the short-term toxicity to fish. The test conducted with the highest substance purity was selected as key study: 96-h LC50 to rainbow trout = 189 mg/L, nominal (OECD 203). The LC50 is far above the water solubility limit of both source and target chemical.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

189 mg/L

Additional information

A study was conducted to determine the toxicity of the structural analogue substance (100% purity) to rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203. The fish were exposed for 96 h to nominal concentrations of 0, 100, 180, 320, 560 and 1000 mg/L. Mortality and symptoms were assessed daily. No analytical verification of test concentrations was conducted. Exposure to 320 mg/L and above induced mortality from 24 h. At the lowest concentration there was 10% mortality at 96 h. Control mortality in one replicate was greater than 10%. However, since there was 10% mortality in the control fish of another test conducted at the same time using the same batch of stock, the overall health of the fish was considered satisfactory. Under the study conditions, the nominal 96 h LC50 was determined to be 189 mg/L (Seaber, 1994).

A study was conducted to determine the toxicity of the test substance (37.3% purity), based on total organic carbon content) to Zebra fish (Danio rerio). Ten fish/group were exposed to the test substance at nominal concentrations of 100, 300 and 1000 mg/L (equivalent to ca. 37, 112 and 373 mg a.i./L respectively). The animals were observed daily for mortality during 96 h. Oxygen, pH and temperature were monitored. No analytical validation of test concentrations was conducted. There was no mortality at 100 mg/L. All fish died within 48 h at 300 mg/L and within 4 h at 1000 mg/L. Under the study conditions, the nominal 96 h LC50 of the test substance was determined to be ca. 149 mg/L (Huntsman, 1986).

A study was conducted to determine the toxicity of the test substance (43% purity) to rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203. The fish were exposed for 96 h to nominal concentrations of 0, 56, 100, 180, 320 and 560 mg/L (equivalent to 0, 20, 40, 72, 128 and 224 mg a.i./L). Mortality and symptoms were assessed daily. No analytical verification of test concentrations was conducted. Exposure to 320 mg/L and above induced mortality from 24 h. At the lowest concentration there was 20% mortality at 96 h. Under the study conditions, the nominal 96 h LC50 of the test substance was determined to be 91 mg/L (Butler, 1993).

A study was conducted to determine the toxicity of the test substance (in the form of aqueous dispersion of 40% purity) to golden orfe (Leuciscus idusL.) according to the guideline DIN 38412. The fish were exposed for 96 h to nominal concentrations of 0, 46.4, 100, 215, 464 and 1000 mg/L (equivalent to 0, 19, 40, 86, 186 and 400 mg a.i./L). Mortality and symptoms were assessed daily. No analytical verification of test concentrations was conducted. Apart from apathy, tumbling and narcotic-like state at 1 and 4 h, no symptoms were observed in any group. Exposure at 215, 464 and 1000 mg/L induced mortality from 4 h. Under the study conditions, the nominal 96 h LC 50 of the test substance was determined to lie between 100 and 215 mg/L (Munk, 1990).

The Seaber (1994) study was conducted with the purest form of the test substance. The other studies were performed with substances partly mixed with dispersants or other chemicals so that exposure through water is artificially increased but not relevant under environmental conditions as the substance is not soluble. Therefore, this study was selected as the key study. Moreover, the Seaber (1994) and Butler (1993) studies were conducted according to OECD 203 compared to the other studies which were conducted as per older guidelines.