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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 March 2010 to 12 March 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
Qualifier:
according to
Guideline:
ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))
Deviations:
no
Qualifier:
according to
Guideline:
other: Guidance document on aquatic toxicity testing of difficult sunstances and mixtures, OECD series on testing and assesment number 23, December 14, 2000.
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Blown rapeseed oil
- Substance type: Yellow to brown viscous liquid
- Physical state: liquid
- Analytical purity: treated as 100% pure
- Expiration date of the lot/batch: 16 December 2010
- Stability under test conditions: stable
- Storage condition of test material: At room temperature in the dark



Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
During the test singular samples for TOC-analysis were taken from the WAF prepared at a loading rate of 100 mg/l and the untreated control according to the schedule below.

Frequency at t=0 h and t=48 h
Volume 40 ml
Storage Samples were stored in a refrigerator until analysis.

Test solutions

Vehicle:
no
Details on test solutions:
The batch of Blown rapeseed oil tested was a yellow/brown viscous liquid with a purity indicated as 100% and not completely soluble in test medium at the concentrations tested.

Preparation of the test solutions started with loading rates prepared individually at 1.0, 10 and 100 mg/l applying 48 hours of magnetic stirring to achieve maximum dissolution in test medium. The magnetic stirring was followed by a settlement period of 2 hours. This resulted in clear and colourless to slightly hazy solutions with a floating layer. The Water Accommodated Fractions (WAFs) were then collected by siphoning the water phase. The WAFs of 1.0 and 10 mg/l were all clear and colourless, while the WAF prepared at 100 mg/l was slightly hazy.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: Daphnia magna (Crustacea, Cladocera) (Straus, 1820)
- Source: In-house laboratory culture with a known history.
- Age at study initiation: < 24 hours
- Method of breeding: With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 10 litres of medium in an all-glass culture vessel.
- Feeding during test: No

Study design

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

Test conditions

Hardness:
180 mg/l expressed as CaCO3
Test temperature:
between 19.6 and 20.4°C
pH:
7.9
Dissolved oxygen:
between 9.0 and 9.5
Nominal and measured concentrations:
Nominal concentrations: control and 100 mg/L
Measured concentrations: Analyses were based on Total Organic Carbon (TOC) contents. The TOC concentrations measured in the treated samples were only slightly exceeding those measured in the control samples, indicating that the actual test substance concentration was very low. The actual measured concentration was at approximately 1.0 to 1.5 mg C/l when corrected for the carbon measured in the control samples. The TOC content of Blown rapeseed oil was determined to be 74.01% (based on NOTOX Project 493030). Thus, the actual Blown rapeseed oil concentration in solution was between approximately 1.4 and 2.0 mg/l.
The low exposure concentration was not unexpected considering the poor solubility of Blown rapeseed oil and the observed undissolved test substance that was present as a floating layer in the dispersion after stirring and which was removed when collecting the water phase for testing.

Details on test conditions:
- Type: open
- Material, size, headspace, fill volume: glass, 100 ml, normal headspace, 80 ml
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for the highest concentration and 2 for the lower concentrations
- No. of vessels per control (replicates): 4
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium: the following chemicals (analytical grade) are dissolved in tap water purified by Reverse Osmosis (RO-water, GEON Waterbehandeling, Berkel-Enschot, The Netherlands):
Macro salts:
CaCl2.2H2O : 211.5 mg/l
MgSO4.7H2O: 88.8 mg/l
NaHCO3 : 46.7 mg/l
KCl : 4.2 mg/l
The hardness: 180 mg/l expressed as CaCO3 and the pH: 7.7 ± 0.3 after aeration.
- Culture medium different from test medium: yes, for culturing M7 medium is used
OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : EC50 t=24, 48 h and NOEC
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Test concentrations: control and 1.0, 10, 100 mg/l
Reference substance (positive control):
yes
Remarks:
potassium dichromate (K2Cr2O7,)

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
100 other: mg/l (WAF prepared at a loading rate of 100 mg/l)
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/l (WAF prepared at a loading rate of 100 mg/l)
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/l (WAF preared at a loading rate of 100 mg/l)
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
Behavioural abnormalities: no
- Immobility of control: no immobility was observed.
- Other adverse effects control: no
Results with reference substance (positive control):
The 24h-EC50 was 0.75 mg/l with a 95% confidence interval between 0.56 and 1.0 mg/l.
The 48h-EC50 was 0.36 mg/l with a 95% confidence interval between 0.28 and 0.49 mg/l.
Reported statistics and error estimates:
No EC50 could be calculated because the test substance proved to be non-toxic (EC50 > maximum soluble concentration).

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the present study Blown rapeseed oil did not induce significant acute immobilisation of Daphnia magna when exposed to a WAF prepared at a loading rate of 100 mg/l after 48 hours of exposure (NOEC). The 48h-EC50 exceeded a WAF prepared at a loading rate of 100 mg/l, which can be considered to represent the maximum soluble concentration in test medium. TOC-analyses showed that the actual concentration in solution was between approximately 1.4 and 2.0 mg/l.