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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:
From July 28,2010 to August 5,2010
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well described GLP compliant study conducted to recognized international test guidelines
according to guideline
EU Method C.2 (Acute Toxicity for Daphnia)
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Fatty acids, C6-24 and C6-24-unsatd., Me esters, distn. residues- Physical state: black , brown semisolid- Analytical purity:100% - Storage condition of test material: room temperature- Solubility: < 10% in water, soluble in acetone, hexane and dichloromethane
Analytical monitoring:
Details on sampling:
Two 2 L beakers were prepared. One was filled with 1L M4 medium for the control.
The following test substance preparation was used:
100 mg + 1000 mL M4 medium
The volume of M4 medium were weighed into one 2L beaker, stirred with magnetic stirrer (300 rpm) and the corresponding amount of the test substance was added to the stirring medium. Preparation was stirred for 24 h.
The test substance preparation was then filtered through a filter paper circle Ø 320 mm (Whatman Schleicher & Schuell GmbH, Dassel, Germany). The control was treated the same way.
Details on test solutions:
Based on a NON-GLP pre-test the test substance C-SAT 100051 was identified as poorly soluble in M4 medium and the test was performed as a limit test with a test substance preparation of 100 mg/L M4 medium.
Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: Daphnia magna
- Strain: Straus
- Source: Landesumweltamt (NRW)
- Age : Daphnia aged less than 24h
- Method of breeding: The daphnids were held in beakers with a nominal capacity of 2000 ml. Each beaker was filled with approximately 1400 ml M4 medium. Between 15 and 20 adult organisms were held in one beaker in order to get a high reproduction rate and viable offspring.
- Feeding during test : yes
- Food type: The algae Desmodesmus subspicatus served as food during holding
- Amount: ad libitum
- Frequency: During holding daphnia were fed from Monday to Friday with an appropriate amount of algae which could be roughly consumed per day. On weekends and public holidays no feeding took place.
- Temperature: 20 ± 2°C
- Dark/light cycle: artificial light-dark cycle of 16 to 8 hours, using fluorescent tubes Osram L36W/21-840 with a light intensity of 700 ± 50 Lux
Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
Post exposure observation period:
The test substance was poorly soluble in M4 medium. After 24 h of stirring the test substance preparation (nominal 100 mg/L) was filtered to eliminate test substance deposits on the surface of the liquid. After filtration the test concentration was present as a clear solution.
2.529 (mmol/L)
Test temperature:
20 ± 2°C
7.94 - 8.05
Dissolved oxygen:
5.6 - 5.9 mg/L
No. Chemical Formula Concentration
1 Sodium nitrate NaNO3 49.6 g/L
2 Dipotassium hydrogenphosphate K2HPO4 3.9 g/L
3 Magnesium sulphate heptahydrate MgSO4 * 7 H2O 7.5 g/L
4 Calcium chloride dihydrate CaCl2 * 2 H2O 3.6 g/L
5 Ferric-III-citrate monohydrate C6H5FeO7 * H2O 0.3 g/L
6 Trace elements
Boric acid H3BO3 2.860 g/L
Manganese chloride tetrahydrate MnCl2 * 4 H2O 1.81 g/L
Zinc sulphate heptahydrate ZnSO4 * 7 H2O 0.220 g/L
Copper sulphate pentahydrate CuSO4 * 5 H2O 0.080 g/L
Sodium molybdate dihydrate Na2MoO4 * 2 H2O 0.024 g/L
Cobalt chloride hexahydrate CoCl2 * 6 H2O 0.040 g/L
7 40 mL of the stock solution 6 were filled up to 1000 mL with double distilled water in a delivery flask. The resulting solution was stock s olution No. 7.
8 Titriplex III Na2 EDTA * 2 H2O 1.0 g/L
9 Citric acid monohydrate C6H8O7 * H2O 0.30 g/L
Nominal and measured concentrations:
Nominal: 100 mg/L
Details on test conditions:
- Test vessel: Five test beakers (50 ml)
- Material, size, headspace, fill volume:
- Aeration: no aeretion
- No. of organisms per vessel: 5
- No. of vessels per concentration :5
- No. of vessels per control:2

- Source/preparation of dilution water:150 L demineralised water was poured into a PVC tank, the 10 solutions were added and mixture was filled up to 160 L with demineralised water and aerated.

Parameter Set range Measured value
Total hardness 2.2 – 3.2 (mmol/L) 2.529
Carbonate hardness 0.4 – 0.7 (mmol/L) 0.455
Calcium hardness 1.2 – 1.8 (mmol/L) 1.584
Magnesium hardness 0.8 – 1.2 (mmol/L) 0.945
pH 7.5 – 8.5 7.53
TOC < 2 mg/L 1.8
conductivity 500 – 700 µS/cm 560
Sodium* 15 - 25 mg/L 18.8
Potassium* 1 - 5 mg/L 3.0
Alkalinity* 0.5 -1.5 mmol/L 0.928

- Adjustment of pH: The pH value adjusted itself to 7.5 – 8.5. The durability of the medium is 4 weeks.
Reference substance (positive control):
potassium dichromate
Key result
48 h
Dose descriptor:
Effect conc.:
> 100 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: 95 % Confidence interval
48 h
Dose descriptor:
Effect conc.:
>= 100 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Details on results:
There was no relevant loss of test substance within 48h as determined by TOC analysis. The EC values given in this report are based on the nominal concentration.
Reported statistics and error estimates:
The experiment was conducted as a limit test; therefore no statistical analysis was necessary.

Validation criteria:


  • There have been no deviations in the environmental test conditions with regard to the pH, temperature and oxygen saturation values (see Appendix 1) whichwere in the required ranges.
  • In the control, not more that 10 percent of the daphnia were immobilized.
Validity criteria fulfilled:
The results of this study showed that the test substance C-SAT 100051 induced no immobilisation to the exposed Daphnia at 100 mg/L within 48 hours. The EC50 value after 48 hours of exposure was >100 mg/L. The NOEC was ≥100 mg/L.
Executive summary:

The test substance C-SAT 100051was tested in the Acute Daphnia Toxicity Test according to the OECD Guideline for the Testing of Chemicals No. 202 (April 2004) usingDaphnia magna(Straus) as test organism.

The experiment was carried out as a limit test (100 mg/L) and as a static test, i.e. the test substance preparation was not exchanged for the whole exposure period of 48 hours.

Description of key information

 EC50  >100 mg/L. 
NOEC ≥100 mg/L.
LC50 (salt water) = 644.61 mg/L
LC0 (salt water)= 100 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
100 mg/L

Additional information

The results of this study showed that the test substance C-SAT 100051 induced no immobilisation to the exposed Daphnia at 100 mg/L within 48 hours. The EC50 value after 48 hours of exposure was >100 mg/L. The NOEC was ≥100 mg/L.

The test was conducted in accordance with the study plan and met all relevant validity criteria. There were no interferences in this test.

Dehylub 1757 displayed a 48h LC50 value of 644.61 mg/L to Acartia tonsa (screening test) in the water phase. The 48h result was calculated by extrapolation, since a 100% effect was not observed at any concentration.