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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1-16 May, 2007
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Study has been performed according to OECD and/or EU guidelines and according to GLP principles.
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
according to guideline
EU Method C.2 (Acute Toxicity for Daphnia)
according to guideline
ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
Details on sampling:
- Concentrations: 0, 10, 22, 46, 100 and 220 mg/l
- Sampling method: at t=0 h and t=48 h 2 ml from the approximate centre of the test vessels. At the end of the exposure period, the replicates were pooled at each concentration before sampling.
- Sample storage conditions before analysis: freezer
Details on test solutions:
Preparation of test solutions started with a solution of 100 (combined limit/range-finding test) or 220 mg/l (final test) applying 15 minutes of magnetic stirring to accelerate the dissolving of the test substance in the test medium. The lower test concentrations (0.1, 1.0 and 10 mg/l in combined limit/range-finding test; 10, 22, 46 and 100 mg/l in final test) were prepared by subsequent dilutions of the 100 or 220 mg/l solution in test medium. The final test solutions were all clear and colourless.
Controls: Test medium without test substance or other additives
Test organisms (species):
Daphnia magna
Details on test organisms:
- Source: In-house laboratory culture with a known history
- Age at study initiation (mean and range, SD): <24 hours
- Method of breeding: With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
- Feeding during test: no feeding

- Type and amount of food: a suspension of fresh water algae
- Feeding frequency: daily
Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
Post exposure observation period:
180 mg CaCO3 per litre

Test temperature:
19.6 - 20.4°C

7.6 - 7.9

Dissolved oxygen:
8.6 mg/l

Not aplicable.
Nominal and measured concentrations:
Only samples taken from the two highest test concentrations and the control were analysed, based on the biological results. Analysis of the samples taken at the start of the test showed measured concentrations of 88 and 213 mg/l at the nominal concentrations of 100 and 220 mg/l, respectively. During the exposure period these measured concentrations remained constant (i.e. 89-91% of initial). Based on these results, the nominal concentrations were used to calculate the effect parameters.
Details on test conditions:
- Test vessel: 100 ml, all-glass
- Aeration: No aeration of the test solutions
- No. of organisms per vessel: 5 per vessel containing 80 ml of test solution
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

- Source/preparation of dilution water:
Medium M7: trace elements, macro nutrients and vitamins are added to freshly prepared ISO medium to reach the following concentrations:
Trace elements: B 0.125 mg/l
Fe 0.05 mg/l
Mn 0.025 mg/l
Li, Rb and Sr 0.0125 mg/l
Mo 0.0063 mg/l
Br 0.0025 mg/l
Cu 0.0016 mg/l
Zn 0.0063 mg/l
Co and I 0.0025 mg/l
Se 0.0010 mg/l
V 0.0003 mg/l
Na2EDTA.2H2O 2.5 mg/l
Macro nutrients: Na2SiO3. 9H2O 10.0 mg/l
NaNO3 0.27 mg/l
KH2PO4 0.14 mg/l
K2HPO4 0.18 mg/l
Vitamins: Thiamine 75.0 µg/l
B12 1.0 µg/l
Biotin 0.75 µg/l
Adjusted ISO medium: the following chemicals (analytical grade) are dissolved in tap water purified by Reverse Osmosis :
Macro salts: CaCl2.2H2O 211.5 mg/l
MgSO4.7H2O 88.8 mg/l
NaHCO3 46.7 mg/l
KCl 4.2 mg/l

- Photoperiod: 16 hours daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : EC50 daily

- Spacing factor for test concentrations: 2.2
- Range finding study: study was a combined limit/range finding test
- Test concentrations: 0.1, 1.0 and 10 mg/l
Reference substance (positive control):
potassium dichromate
Key result
48 h
Dose descriptor:
Effect conc.:
> 220 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Details on results:
Combined limit/range finding test:
After 48 hours of exposure 60% immobility was observed at 100 mg/l. No significant immobility was observed at any of the lower test concentrations during the test period. Therefore, the expected EC50 was just above nominal 100 mg/l.
Immobility at 48h: 10% at 0.1 mg/l; 0% at 1.0 mg/l; 10% at 10 mg/l; 60% at 100 mg/l.

Final test:
No significant immobility was observed at any of the test concentrations during the test period. Therefore, the responses recorded at nominal 100 mg/l were not in agreement with the results of the combined limit/range-finding test. However, the responses recorded in this final test allowed for reliable determination of an EC50 because the results at 220 mg/l supported the results at 100 mg/l.
Immobility at 48h: 0% at 10 mg/l; 5% at 22 mg/l; 0% at 46 mg/l; 5% at 100 and 220 mg/l.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Mortality at 48h: 0% at 0.10 mg/l; 5% at 0.18 mg/l; 50% at 0.32 mg/l; 85% at 0.56 mg/l; 100% at 1.0 and 1.8 mg/l
- EC50/LC50: 48h-EC50 was 0.34 mg/l with a 95% confidence interval between 0.28 and 0.40 mg/l.
Validity criteria fulfilled:
Under the conditions of the present study Ditetrahydrofurylpropane did not induce significant acute immobilisation of Daphnia magna at 220 mg/l after 48 hours of exposure (NOEC).
The 48h-EC50 was beyond 220 mg/l based on analytically confirmed nominal concentrations.

Description of key information

A study was conducted according to OECD 202. No significant immobility was observed at any of the test concentrations during the test period.

Key value for chemical safety assessment

Additional information

The 48h-EC50 was beyond 220 mg/l based on analytically confirmed nominal concentrations.