reaction mass of: tetrasodium 4-amino-6-(5-(2,6-difluoropyrimidin-4-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(sulfatoethylsulfonyl)phenylazo)naphthalene-2,7-disulfonate;tetrasodium 4-amino-6-(5-(4,6-difluoropyrimidin-2-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(2-sulfatoethylsulfonyl)phenylazo)naphthalene-2,7-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:

1998-03-02 to 1998-05-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Principles of method if other than guideline:

None

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/L and blank control
- Sampling method: water samples taken from the middle of the test chamber by pipetting approx 20 ml in to a screw neck glass bottle
- Sample storage conditions before analysis: No data

Vehicle:

no

Details on test solutions:

Identity and concentration of auxiliary solvent for dispersal: None
Preparation details: Once the water had been added to the test substance, the mixture was homogenised using an Ultra Turrax and poured into the test chamber under stirring with a glass rod.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish (Danio rerio Hamilton-Buchanan)
- Batch number: 16a/97/L2a
- Strain: not applicable
- Source: Hoechst Marion Roussel Deutschland GmbH Global preclinincal Development - Drug Safety
- Age at study initiation: > 71 days old at start of study (details not included in study report)
- Length at study initiation = mean 2.7 cm, variation 2.7-2.9, SD = 0.09 cm

ACCLIMATION
- Acclimation period: Stock held since 11 December 1997 and acclimated to for 15 days.
- Acclimation conditions
- Temp. = 22 ± 1 °C
- Type and amount of food: tetra Min, tetra Werke, twice daily
- Health during acclimation (any mortality observed): Not stated
- Population density: ≤1g fish/L water

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

2.1 mmol/L (Ca2+ and Mg2+)

Test temperature:

21.3-21.7 °C (during test)

pH:

7.5-8.0 (during test)

Dissolved oxygen:

7.4-10.1 mg/L during study

Salinity:

N/A

Nominal and measured concentrations:

Nominal: 100 mg/L and 0 mg/L (control)
Measured concentrations: 96.8-97.1 mg/L and <0.5 mg/L (control)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria holding 4 L of test media
- Aeration: not aerrated during the course of the study
- Renewal rate of test solution (frequency/flow rate): None
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water prepared according to the recommendations of ISO 7346
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Photoperiod: 12 light/12 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Mortality. In addition subjective assessments were made on the type and incidence of sub-lethal effects compared with control fish, which included recording of visible changes in appearance and behaviour. Assessments were made at 3, 6, 24, 48, 72 and 96 hours.

TEST CONCENTRATIONS
- Range finding study: Not carried out
- Test concentrations: 100 mg/L

Reference substance (positive control):

no

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)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality was observed at any point in time in either in the exposed or the control fish.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

None

Validity criteria fulfilled:

yes

Conclusions:

The 96 hour LC50, LC0 and NOEC of the test substance was determined to be > 100 mg/L (analytically verified).

Executive summary:

A study was performed to assess the acute toxicity of test substance to Danio rerio (zebra fish) under static conditions. The study was conducted in accordance with EU Method C.1 and in compliance with GLP. Groups of seven fish of the recommended size were exposed to a limit test substance concentration of nominally 100 mg/L. The test substance was directly weighed to the test medium and no auxiliaries were used. The test substance was measured in the water at 96.97 mg/L. A control was also included (concentration <0.5 mg/L, limit of detection).

Observations were made on the number of dead fish and the incidence of sublethal effects after 3, 6, 24, 48, 72 and 96 hours of exposure. No mortality and no sublethal effects were observed in any of the test fish at any point in time. Therefore, under the employed test conditions the LC50 value at any time from 0-96 h was > 100 mg/L. An LC50 cannot be established, however it will be >100 mg/L.

This toxicity study is classified as acceptable and satisfies the guideline requirement.

Description of key information

The 96 hour LC50 value of the test substance was determined to be >100 mg/L (measured test substance). No mortality and no sublethal effects were observed in any of the test fish at any points in time. Therefore, no LC50 can be established.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

A study was performed to assess the acute toxicity of the test substance to Danio rerio (zebra fish) under static conditions. The study was conducted in accordance with EU method C.1 and in compliance with GLP. Groups of seven fish of the recommended size were exposed to a limit test substance concentration of nominally 100 mg/L. The test substance was directly weighed into the test medium and no auxiliaries were used. The test substance concentration in water was analytically measured at 96.97 mg/L. A control was included (concentration <0.5 mg/L).

Observations were made on the number of dead fish and the incidence of sublethal effects after 3, 6, 24, 48, 72 and 96 h of exposure. No mortality and no sublethal effects were observed in any of the test fish at any point in time. Therefore, under the employed test conditions the LC50 value at any time from 0-96 h was > 100 mg/L.

This toxicity study is classified as acceptable and satisfies the guideline requirement.