reaction product of diphenylmethanediisocyanate, octylamine, 4-ethoxyaniline and ethylenediamine (1:0,37:1,53:0,05)

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:

1999-10-23 to 1999-02-15

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: No concentrations in excess of the water solubility limit of the test substance were tested. Due to the low solubility of the test substance in test water the following dosage was chosen: A supersaturated stock suspension of the test substance with a nominal concentration of 100 mg/L was prepared by dosing 500 mg of the test substance into 5 litre test water. No auxiliary solvent or emulsifier was used. The test substance was mixed into the test water as homogeneously as possible by ultrasonic treatment for 15 minutes and intense stirring. The supersaturated stock suspension was stirred on a magnetic stirrer at room temperature in the dark over 48 hours to dissolve respectively disperse a maximum concentration of the test substance in the stock suspension. Due to the results of a pre-experiment no differences of dissolved amount of test substance in test water were determined after stirring periods between 2 and 48 hours. The stock suspension of the test substance was filtered through a cellulose-nitrate filter with a defined pore size of 0.2 µm just before the start of the test. Due to this procedure it was seen to that the substance was solved in the test water up to the solubility limit. All particles of the test substance up to a size of 0.2 μm were solved or suspended in the test water. Thus, it was guaranteed that the substance was tested at the maximum solubility limit. No concentration above the solubility limit of the test substance in the used test water were tested to avoid physical effects of undissolved test substance onto the test animals. Additionally, a control was tested in parallel. Thus, a limit test was performed to demonstrate that the test substance has no toxic effect on fish up to the solubility limit in the used test water. The only concentration tested was a filtrate of a supersaturated stock suspension of nominal 100 mg test substance/L and a control.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Source: The test fish were obtained from City Zoo, 64283 Darmstadt, Germany.
- Age at study initiation: juveniles
- Length at study initiation: 3.0 ± 0.13 cm
- Weight at study initiation: 0.20 ± 0.03 g

ACCLIMATION
- Acclimation period: 5 weeks
- Type and amount of food: TETRA MIN Hauptfutter, TETRA-Werke, D-49324 Melle, Germany
- Feeding frequency: During holding until one day before test start the fish were fed with a commercial fish diet
- Health during acclimation: During the 5 weeks prior to the test no fish died in the test fish batch and all fish were healthy

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No

Hardness:

Not specified

Test temperature:

21 - 22 °C

pH:

7.6 to 7.9

Dissolved oxygen:

at least 8.0 mg/L

Salinity:

Not applicable

Conductivity:

Not specified

Nominal and measured concentrations:

100 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: 9-litre glass aquaria with 5 litre test medium
- Aeration: The test media were slightly aerated during the test.
- No. of organisms per vessel: 7 fish were introduced in each aquarium
- No. of vessels per concentration: one vessel
- No. of vessels per control: one vessel

TEST MEDIUM / WATER PARAMETERS
- in accordance to the guideline

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16 h light: 8 h dark
- Light intensity: 213 - 248 lux

EFFECT PARAMETERS MEASURED:
The test fish were observed after approximately 3, 24, 48, 72 and 96 hours test duration for symptoms of intoxication and mortality.

Reference substance (positive control):

no

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:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

All fish survived until the end of the test and no signs of intoxication were observed.

Results with reference substance (positive control):

No positive control performed (not applicable).

Reported statistics and error estimates:

Not applicable.

Validity criteria fulfilled:

yes

Conclusions:

The test item has not toxic effects to fish up to a nominal concentration of 100 mg/L after 96 hours of exposure.

Executive summary:

The acute toxicity of the test substance UREA 4 to Zebra fish (Brachydanio rerio) was determined in a 96-hour static test. Due to the low water solubility limit of the test substance a filtrate of a supersaturated stock suspension of nominal 100 mg/L was tested. Thus, no concentrations above the solubility limit of the test substance in the used test water were tested. Additionally, a control was tested in parallel. Thus, a limit test was performed to demonstrate that the test substance has no toxic effect on fish up to the solubility limit in the used test water. Due to the low water solubility limit no analytical dose verification could be done in the present test. Therefore, the biological results can not be related to a specific concentration of the test substance but to the solubility limit in the test medium. The 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), respectively the 96-hour LC 0 of UREA 4 to Zebra fish was determined to be at least up to the solubility limit of the test substance/L in test water. The NOEC and the LC 0 might even be higher than this concentration, but concentrations in excess of the solubility limit have not been tested. The 96-hour LOEC, the 96-hour LC 50 and the 96-hour LC 100 were clearly higher than the solubility limit of the test substance in test water. These values could not be quantified due to the absence of toxicity of UREA 4 up to the tested concentration.

Description of key information

The acute toxicity to fish was assessed under static conditions to a nominal concentration of 100 mg/L. The LC50 was calculated to be > 100 mg/L. The NOEC was greater or equal 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The acute toxicity of the test substance UREA 4 to Zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to EU method C.1 and OECD guideline 203. Due to the low water solubility limit of the test substance a filtrate of a supersaturated stock suspension of nominal 100 mg/L was tested. Thus, no concentrations above the solubility limit of the test substance in the used test water were tested. Additionally, a control was tested in parallel. Thus, a limit test was performed to demonstrate that the test substance has no toxic effect on fish up to the solubility limit in the used test water. Due to the low water solubility limit no analytical dose verification could be done in the present test. Therefore, the biological results can not be related to a specific concentration of the test substance but to the solubility limit in the test medium. The 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), respectively the 96-hour LC 0 of UREA 4 to Zebra fish was determined to be at least up to the solubility limit of the test substance/L in test water. The NOEC and the LC 0 might even be higher than this concentration, but concentrations in excess of the solubility limit have not been tested. The 96-hour LOEC, the 96-hour LC 50 and the 96-hour LC 100 were clearly higher than the solubility limit of the test substance in test water. These values could not be quantified due to the absence of toxicity of UREA 4 up to the tested concentration.