Reaction mass of pentasodium 2-{[4-chloro-6-(ethyl{3-[(2-sulfonatoethyl)sulfonyl]phenyl}amino)-1,3,5-triazin-2-yl]amino}-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate and tetrasodium 2-[(4-chloro-6-{ethyl[3-(vinylsulfonyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate

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

Study period:

24 March 2006 to 26 June 2006

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:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40824/A
Batch: Red ROE 805 BOP 04/05
Appearance: dark red powder
Purity: Organic part (Na-salt): approx. 82 %; Main component 1 : approx. 36.2 %; Main component 2: approx. 27.5 %; Oligomers: 10%
Expiration date: 01 October 2010
Stability in water: Max. 7 days at room temperature
Solubility in water: >100g/Lat20°C
Storage: At room temperature at about 20 °C, in a desiccator because test substance is hygroscopic, away from direct sunlight

Analytical monitoring:

yes

Details on sampling:

Duplicate samples from each test medium and the control just before the start of the test and after 48 and 96 hours.
All samples were taken from the approximate center of the aquaria without mixing of the test media, and were deep-frozen (at about -20 °C) immediately after sampling.

Vehicle:

no

Details on test solutions:

The following nominal concentrations were tested: 1.0, 3.2, 10, 32 and 100 mg/L.
A control was tested in parallel .
The test medium of the highest nominal test concentration of 100 mg/L was freshly prepared by dissolving 750.2 mg of the test item completely in 7500 mL of test water using stirring for 15 minutes at room temperature.
Adequate volumes of the test medium of the highest test concentration were added to the test water in the aquaria and were intensively mixed to prepare the test media with the lower test concentrations.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Test organism: Zebra fish (Brachydanio rerio)
Weight: 0.21 ± 0.05 g.
Length: 3.0 ± 0.2 cm

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

125 mg/L as CaCO3

Test temperature:

21 - 22 °C

pH:

7.3 - 7.4

Dissolved oxygen:

8.1 - 8.6 mg/L

Salinity:

No data

Nominal and measured concentrations:

nominal: 1.0, 3.2, 10, 32 and 100 mg/L
measured: between 100 and 107 % of the nominal values at the start and the end of the test

Details on test conditions:

Volume: 5 liters test medium
Size: 7 fish per glass aquarium
Aeration: Slowly aerated
Light condition: A 16-hour light to 8-hour dark photoperiod, with a 30-minute transition period (light intensity during the light period was approximately within the range of 50 to 500 Lux).

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

32 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)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality occurred in the control and in the concentration of 1.0,3.2,10 and 32 mg/L.
At the concentration of 100 mg/L, one of the seven test fish was dead at the end of the test.

Results with reference substance (positive control):

No data

Reported statistics and error estimates:

These values could not be quantified due to low toxicity of FAT 40824/A at the tested concentrations.

Sublethal observations / clinical signs:

The analytically determined concentrations of the test item FAT 40824/A in the analyzed test media were between 100 and 107% of the nominal values at the start and the end of the test. The test item was stable during the test period of 96 hours under the test conditions. The biological results were based on the nominal concentrations of the test item.

In the control and in the test media up to and including the test item concentration of 32 mg/L, all fish survived until the end of the test and no visible abnormalities were observed. At the highest test item concentration of 100 mg/L, all test fish showed symptoms of toxicity after 96 hours test duration and one of the seven test fish was dead at the end of the test.

Thus, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours) and the 96-hour LC0 of FAT 40824/A to zebra fish were determined to be 32 mg/L. The 96-hour LOEC (lowest concentration tested with observable toxic effects) was determined to be 100 mg/L. The 96-hour LC50 and the 96-hour LC100 were clearly higher than 100 mg/L. These values could not be quantified due to low toxicity of FAT 40824/A at the tested concentrations.

No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions colored by the test item throughout the entire test duration.

The pH values in the test media and the control ranged from 7.3 to 7.4. The oxygen concentration was always 8.1 mg/L or higher, and thus higher than 60% oxygen saturation. The water temperature was between 21 and 22 °C.

Validity criteria fulfilled:

yes

Conclusions:

The NOEC and LC0 of FAT 40824/A was 32 mg/L while LOEC was 100 mg/L. The LC50 and the LC100 were higher than 100 mg/L.

Executive summary:

A GLP compliant acute toxicity of FAT 40824/A to zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to the OECD TG 203(1992) and EU 92/69/EEC, C.1 (1992). The nominal test item concentrations tested were 1.0, 3.2, 10, 32 and 100mg/L in parallel with a control. From the acclimated test fish batch, 10 fish were measured at the start of the test: The mean body length of the fish was 3.0 ± 0.2 cm. The mean body wet weight was 0.21 ± 0.05 g.

No mortality occurred in the control and in the concentration of 1.0, 3.2, 10 and 32 mg/L at the end of test. Based on the test results, the NOEC and LC₀ were 32 mg/L. The LOEC was 100 mg/L. The LC₅₀ and the LC₁₀₀ were higher than 100 mg/L.

Description of key information

The NOEC and LC0 were 32 mg/L. The LOEC was 100 mg/L. The LC50 and the LC100 were higher than 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

A GLP compliant acute toxicity of FAT 40824/A to zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to the OECD TG 203(1992) and EU 92/69/EEC, C.1 (1992). The nominal test item concentrations tested were 1.0, 3.2, 10, 32 and 100mg/L in parallel with a control. From the acclimated test fish batch, 10 fish were measured at the start of the test: The mean body length of the fish was 3.0 ± 0.2 cm. The mean body wet weight was 0.21 ± 0.05 g.

No mortality occurred in the control and in the concentration of 1.0, 3.2, 10 and 32 mg/L at the end of test. Based on the test results, the NOEC and LC₀ were 32 mg/L. The LOEC was 100 mg/L. The LC₅₀ and the LC₁₀₀ were higher than 100 mg/L.