Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02 Apr - 03 May 2009
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Due to the nature of the test item, analytical measurement is not possible.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Qualifier:
according to
Guideline:
other: OECD No.23 Guidance document on aquatic toxicity testing of difficult substances and mixtures
GLP compliance:
yes (incl. certificate)
Remarks:
THE DEPARTMENT OF HEALTH OF THE GOVERNMENT OF THE UNITED KlNGDOM, UK GLP Monitoring Authority

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
no
Remarks:
Due to the nature of the test item, analytical measurement is not possible.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Differential loading: Since a limittest was conducted the only test item concentration was 100 mg/L. The test solution was prepared by stirring the appropriate amount of test material in 20 L of test media for 48 h at 21 - 23 °C. At the end of the stirring phase the dispersion was settled for 4 h. The aqueous phase was seperated from the bulk of the undissolved material and filtered using a Whatman No.1 filter paper. The resultant solution was used as the test exposre solution.
- Controls: Medium without test item

Test organisms

Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: Common carp
- Source: Hampshire Carp Hatcheries; Bow Lake Fish Farm, Hampshire, UK
- Mean length at the end of the study: 56.2 mm
- Mean weight at the end of the study: 1.79 g

ACCLIMATION
- Acclimation period: 20 - 29 Apr 2009
- Acclimation conditions: Same as test conditions:Temperature: 21.5 - 23.5 °C; Dissolved oxygen: >81% ASV; Photoperiod: 16 h light and 8 h dark
- Type and amount of food during acclimation: The carp are fed with Hikari Staple carp food approximately 2% body weight per day until 24 h before test initiation.
- Feeding frequency during acclimation: daily
- Health during acclimation (any mortality observed): 3.3% mortality during acclimation period

FEEDING DURING TEST : The carp were not fed during the test

Study design

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

Test conditions

Hardness:
183 mg CaCO3/L
Test temperature:
21.0 - 22.5 °C (Control)
21.0 - 23.0 (100 mg/L)
pH:
7.7 - 8.1 (Control)
7.9 - 8.1 (100 mg/L)
Dissolved oxygen:
75 - 100% ASV (Control)
86 - 99% ASV (100 mg/L)
Nominal and measured concentrations:
Control and 100 mg/L (nominal loading rate)
Details on test conditions:
TEST SYSTEM
Test vessel:
- Type: open
- Material, size, headspace, fill volume: 28 L volume aquaria, 20 L test volume, 8 L headspace
- Renewal rate of test solution (frequency/flow rate): every 24 h
- 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: Dechlorinated main water with reverse osmosis purified water added appropriately to lower the hardness. The ratio of dechlorinated mains water to reverse osmosis water was 75:25. The hardness was calculated after mineral analysis on a sample of dilution water.
- Culture medium different from test medium: same medium was used
- Intervals of water quality measurement: The pH (to 0.1), dissolved oxygen (to 1% ASV) and temperature (to 0.5 °C) were measured. Water quality determinations for each test and control solution were carried out immediately prior to initiating the test, before and after each daily change of the test solutions and at the end of the 96 h exposure period.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h dark

EFFECT PARAMETERS MEASURED: Observations and records of mortalities were made after 24, 48, 72 and 96 h exposure period.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: no spacing facter was used since only one test item concentration was tested
Range finding study
- Test concentrations: 0, 0.1, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study data from the preliminary study identified the 96 h LL50 as being greater than 100 mg/L.
Reference substance (positive control):
yes
Remarks:
Copper (II) Sulphate (CuSO4・5H2O)

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:
- Behavioural abnormalities: No abnormal behaviour was observed.
- Mortality of control: No mortality was observed.
Results with reference substance (positive control):
LC50 (96h): 4.6 mg/L (95% confidential limits: 2.9 - 7.2 mg/L)
Reported statistics and error estimates:
All LC50 and NOEC results are calculated using ToxCalc Version 5.0 "Comprehensive Toxicity Data Analysis and Database Software"

Any other information on results incl. tables

Biological Results:

The 96 h LL50 value of the test item to Carp was estimated to be greater than 100 mg/L. The highest no observed effect loading rate (NOELR) after 96 h was 100 mg/L.

Table 1: LL50 and NOELR values (based on the nominal loading rate)

Period of exposure (h)

LL50 value

95% confidence limits

NOELR

24

>100 mg/L

Not possible to determine

100 mg/L

48

>100 mg/L

Not possible to determine

100 mg/L

72

>100 mg/L

Not possible to determine

100 mg/L

96

>100 mg/L

Not possible to determine

100 mg/L

Table 2: Abnormal behavior

Period of exposure (h)

Nominal loading rate (mg/L)

 

Control

100

0

7 n

7 n

24

7 n

7 n

48

7 n

7 n

72

7 n

7 n

96

7 n

7 n

N: number of carp displaying normal behaviour

Table 3: Cumulative mortalities

Period of exposure (h)

Nominal loading rate (mg/L)

 

Control

100

0

0

0

24

0

0

48

0

0

72

0

0

96

0

0

Table 4: Validity criteria

Criterion from the guideline

Outcome

Validity criterion fulfilled

The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test

No mortality was observed

Yes

The dissolved oxygen concentration must have been at least 60 per cent of the air

saturation value throughout the test

Oxygen concentration ≥ 60%

Yes

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
For further details please refer to “Any other information on results incl. tables”.