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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From October 05 to 07, 2000.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
This study was performed according to international guidelines with GLP statement. All validity criteria were fulfilled but the analytical determination was not specific (DOC analysis) and was performed only at the start of the test.
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
. The analytical determination was not specific (DOC analysis) and was performed only at the start of the test.
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
yes
Remarks:
. The analytical determination was not specific (DOC analysis) and was performed only at the start of the test.
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Remarks:
1997-07-07
Specific details on test material used for the study:
No additional information
Analytical monitoring:
yes
Details on sampling:
The test item was analytically verified at test begin.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Saturated solution (50 mg/L were weighed out), prepared with dilution water one day prior to application (24h on a magnetic stirrer) and filtered (0.45 µm) before use.
- Eluate: dilution water
- Controls: One control. Dilution water without test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable
- Concentration of vehicle in test medium (stock solution and final test solution): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None. The test item solution was clearly dissolved throughout exposure.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna STRAUS
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- und Lufthygiene des Bundes-gesundheitsamtes, Corrensplatz, D-14195 Berlin.
- Age at study initiation (mean and range, SD): less than 24h old
- Method of breeding: Daphnia types are bred in the test facility. In 2-3 L glass vessels with approximately 1.8 L culture medium, by 21°C (temperatures of 20-25°C were tolerated), in an incubator, 16h illumination, illumination strength 1.5-5.0 µmol.m^-2.s-1.
- Feeding during test: no data
- Feeding: 5 x weekly ad libitum with a mix of Scenedesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 10^6 cells/mL.

ACCLIMATION
2h in dilution water
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Remarks on exposure duration:
None
Post exposure observation period:
None
Hardness:
227 mg/L as CaCO3.
Test temperature:
21 +/- 1°C
pH:
7.32-7.72
Dissolved oxygen:
8.26-8.39 mg O2/L
Salinity:
Not applicable
Nominal and measured concentrations:
- Nominal concentration: 50 mg/L
- Measured concentration: saturated solution, 0.92 mg/L. See Table 6.1.3/1 in "Any other information on results incl. tables".
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (5 cm ID x 8 cm H), fill volume 50 mL, volume of the study medium 20 mL.
- Aeration: no data
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): not applicable
- No. of organisms per vessel: 5 (10 for the reference item)
- No. of vessels per concentration (replicates): 4 (2 for the reference item)
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): not applicable

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: M4, ELENDT (1990)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8h light/dark cycle
- Light intensity: Diffuse light, illumination range 1.5-5 µmol.m^-2.s^-1.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation

TEST CONCENTRATIONS
- Spacing factor for test concentrations: None (limit test, saturated solution)
- Range finding study: A preliminary range-finding test was conducted under static conditions with three dilutions of the saturated solution (1:1, 1:10, 1:100, NON-GLP state). For the stock solution 100 mg/L were weighed out.
- Results used to determine the conditions for the definitive study: No biological significant effect (<= 10 %) was seen in the saturated solution.

Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 0.92 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks:
corresponding to a loading rate of 50 mg/L
Remarks on result:
other: no observed effect up to the attainable test item concentration
Duration:
48 h
Dose descriptor:
other: The highest concentration without observed effect
Effect conc.:
>= 0.92 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks:
corresponding to a loading rate of 50 mg/L
Remarks on result:
other: no observed effect up to the attainable test item concentration
Details on results:
- Behavioural abnormalities: none
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: none

See results of the definitive test in the Table 6.1.3/2 in "Any other information on results incl. tables".
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- 24h-EC50 = 2.0 mg/L (CL: 1.8-2.1 mg/L)
Reported statistics and error estimates:
None

Table 6.1.3/1: DOC - Analysis of new media (0 h)

 

DOC measured (mg/L)

Test item concentration*

(mg/L)

Saturated solution

0.75

0.92

Control

0

-

* calculated based on ThTOC of 0.813 mgC/mg.

Table 6.1.3/2: Percentage number of Daphnids incapable of swimming after 24 and 48 h of exposure (n = 20)

Immobilisation (%)

Test item concentration (mg/L)

24h

48h

Replicates

Replicates

1

2

3

4

Mean value

1

2

3

4

Mean value

0.92

Control

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Validity criteria fulfilled:
yes
Conclusions:
No biological significant effect was observed up to the limit of solubility. The 48h-EC50 was > 0.92 mg/L (saturated solution).
Executive summary:

This study was performed according to OECD Guideline 202 and EU Method C.2 with GLP statement, to assess the 48h-acute toxicity of the saturated solution of the test substance to Daphnia magna, under static conditions.

In this study, a limit test was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effect on the daphnids up to the attainable test substance concentration. Due to the low water solubility of the test item, a saturated solution of the test substance with 50 mg/L was prepared and was continuously stirred over 24 hours. Then, the dispersion was filtered through a 0.45 μm membrane filter. The undiluted filtrate of the dispersion was used as the test medium. Additionally, a control was tested in parallel. 

20 test organisms were exposed to the limit test concentration and control. The test item concentration was analytically verified using DOC analysis according to DIN 38409 part 3, at the start of the test.

In the saturated solution, at 0.92 mg/L, no biologically significant effect was determined.

A reference test is carried out once per month with potassium dichromate as reference item and the reference toxicity determined. The EC50 -value of the reference item at 2.0 mg/L after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/L according to AQS, DIN Guideline 38412 L 30.

Water quality parameters of pH-value and dissolved oxygen concentration measured at 0 and 48 h were determined to be within the acceptable limits.

All validity criteria were fulfilled.

Description of key information

EU Method C.2, OECD Guideline 202, GLP, key study, validity 2:

No acute effect up to the attainable limit of solubility in test medium.

48h-EC50 > 0.92 mg/L., corresponding to a loading rate of 50 mg/L.

Key value for chemical safety assessment

Additional information

To assess the short-term toxicity of the registered substance, (-)-(3aR,5aS,9aS,9bR)-3a,6,6,9a-Tetramethyldodecahydronaphtho[2,1-b]furan, to aquatic invertebrate Daphnia magna, two studies are available.

The first study (Noack, 2000), assessed as the key study, was performed on the registered substance according to OECD Guideline 202 and EU Method C.2 with GLP statement. Daphnids were exposed to a saturated solution of the substance, under static conditions. Due to the low water solubility of the substance, a saturated solution with 50 mg/L of the test substance was prepared, continuously stirred over 24 hours, filtered through a 0.45 µm membrane filter, and then the undiluted filtrate of the dispersion was used as the test medium. The measured concentration was analysed only at the start of the test, at 0.92 mg/L. No biologically significant effect was determined during the test period, therefore the 48h-EC50 value was greater than the maximum initial solubility concentration of 0.92 mg/L. The test substance had no toxic effects on Daphnia magna up to its water solubility limit in test medium at a loading rate of 50 mg/L.

The second study (Henkel, 1993), assessed as a disregarded study, was performed on the registered substance according to German guideline DIN 38412 part 11 with GLP statement. This study was not considered valid because the results were based on nominal concentrations while the measured concentrations were well lower than nominal and the substance floating on the surface, therefore the effects observed can be related to physical effects.