Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 - 30 Nov 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Niedersächsisches Umweltministerium, Hannover, Germany

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: control, 100 mg/L
- Sampling method: Samples were taken from the test solutions and an additional replicate without test organisms (duplicate samples).

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 200 mg test substance was applied to 2 L of test medium. The solution was shaken with 20 rpm for 24 h. Thereafter, the dispersion was centrifuged (20 min, 3000 rpm) to remove undissolved particles.
- Eluate: no
- Differential loading: no
- Controls: yes, test medium control

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebrafish
- Source: Aquarium am Aegi, Hannover, Germany
- Maintenance of the brood fish: Holding was performed at 23 ± 2 °C and diffuse light. Water was changed at least once per week. No disease treatments were administered throughout holding and testing.
- Length at study initiation: 2.67 cm
- Weight at study initiation: 0.2 g

ACCLIMATION
- Acclimation period: at least 12 d (mortality < 5% within the last 7 d of acclimatisation)
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: Trouvit 40 / 2, Milkivit; Bürgheim, Germany
- Feeding frequency during acclimation: Three times a week (4% of fish body weight per day); Fish were not fed until 24 h before test start.

FEEDING DURING TEST
- None

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
76 mg/L as CaCO3
Test temperature:
21.0 - 22.5 °C
pH:
7.04 - 7.53
Dissolved oxygen:
100% oxygen saturation
Nominal and measured concentrations:
nominal: control, 100 mg/L
measured DOC: please refer to "Any other information on results incl. tables"
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass aquaria with test volume of 2 L per vessel.
- Type (delete if not applicable): loosely covered by glass tops
- Aeration: gentle aeration was provided
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 1 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water of local origin; water was filtered on activated charcoal and aerated for at least 24 h to remove chlorine
- Total hardness: 40 - 180 mg CaCO3/L
- pH: 6.0 - 8.0
- Culture medium different from test medium: same as test

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: natural photoperiod
- Light intensity: 0.1 - 10 µmol/m²*s

EFFECT PARAMETERS MEASURED
- Mortality was recorded after 24, 48, 72 and 96 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable, limit test
- Range finding study
- Test concentrations: One limit Water Accommodated Fraction (WAF) was tested incl. a control
- Results used to determine the conditions for the definitive study: No mortality was recorded. Thus, a limit test is justified.
Reference substance (positive control):
not required

Results and discussion

Effect concentrations
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of control: 0%
Reported statistics and error estimates:
Since no effects were recorded, no statistical analysis was necessary.

Any other information on results incl. tables

Table 1: Results from DOC analysis

Nominal loading [mg/L]

DOC [mg/L]

Parallel determination

Mean

0 h

96 h

0 h

96 h

100

--

--

46.5

47.0

--

46.8

100*

43.3

43.5

42.8

43.1

43.4

43.0

Control

--

--

6.4

6.5

--

6.5

Control*

1.6

1.6

1.7

1.8

1.6

1.8

* test loading without test organisms

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
Mortality in control: 0%; O2 saturation: 100%