Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Nov - Dec 1993
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
31 July 1992
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
Acid Red 299

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 10, 29, 83 mg/L
- Sampling method: the samples were taken from the middle of the water body using pippetes

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
stock solution: The test item (1000mg) were dissolved in 250mL distilled water.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish (Danio rerio)
- Source: ASCANIA-AQUARIUM, Aschersleben, Germany
- Age at study initiation: juvenile fish
- Length at study initiation (mean): 33.7mm
- Mortality of test population: <5%

ACCLIMATION
- Acclimation period: minimum 7 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: standard food daily until 2 days before commencement of test

FEEDING DURING TEST
- Food type: standard food (pestled trout fish food)
- Frequency: daily until 2 days before commencement of test

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
c(Ca++ and Mg++) = 2.5mmol/L
Test temperature:
22.7-23.1°C
pH:
pH Control: 8.0
pH Test: 8.0 - 8.2
Dissolved oxygen:
88-95%
Nominal and measured concentrations:
nominal: 0, 10, 17, 29, 49, 83 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: aquarium
- fill volume: 4L
- Aeration: yes
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 1

OTHER TEST CONDITIONS
- Adjustment of pH: no
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
29 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
49 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
37.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: LC50 as geometric mean between LC0 and LC100
Details on results:
- Behavioural abnormalities:
- Observations: animals of test group no: 1 a light pink coloring was observed
- Other biological observations: animals of test group no: 2 and 3 were slightly damaged and swam during the first hour on the water surface and reacted oversensitive to external stimuli. At the end of the exposure a reddish coloring on the animales was observed.
- Due to the intense coloring of the test solution no observation could be made in respect of test group 4 and 5.
- At the end of the exposure period a dark, flaky precipitation was observed on the bottom of the test vessels (increased with higher concentrations). These is probably caused of the excrements of the fish.
- Mortality of control: no
- Other adverse effects control: none observed

Any other information on results incl. tables

            Exposure Time
 Conc. (mg/L)  24 (hours)  48 (hours)  72 (hours)  96 (hours)
 LC0  29  29  29  29
 LC100  49  49  49  49

Mortality

              Live animals after:  
 Test Group No.  Conc. (mg/L)  24 h  48 h  72 h  96 h  Mortality Rate after 96 h (%)
 0 (Control)  0  7  7  7  7  0
 1  10  7  7  7  7  0
 2  17  7  7  7  7  0
 3  29  7  7  7  7  0
 4  49  0  -  -  -  100
 5  83  0  -  -  -  100

pH values during exposure

    Test Group     pH values during exposure (hours)
 No.  Conc. (mg/L)  2  24  48  72  96
 0  0  8,0  8,0  8,0 8,0   8,0
 1  10  8,0  8,0  8,0  8,0  8,0
 2  17  8,0  8,0  8,0  8,0  8,0
 3  29  8,1  8,0  8,0  8,1  8,1
 4  49  8,1  8,1  -  -  -
 5  83  8,2  8,1  -  -  -

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The 96-h LC50 to Danio rerio was determined to be > 29 and < 49mg/L. The calculated LC50 (as geometric mean value between LC0 and LC100) is 37.7 mg/L. The substance is acutely harmful to fish.