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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 October 2009 - 20 October 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This 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:
2009
Report date:
2009

Materials and methods

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

Test material

Constituent 1
Reference substance name:
Biomass residue (liquid) obtained after B2 production
IUPAC Name:
Biomass residue (liquid) obtained after B2 production

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Samples for Total Organic carbon (TOC) analysis were taken from the highest test concentration and the 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 Biomass residue (liquid) obtained after B2 production tested was a brownish-yellow turbid liquid containing 25-35% dry matter in water. Although the test substance was indicated to be soluble in water, it was not completely soluble in ISO-medium at loading rates of 10 and 100 mg/l (based on the liquid suspension as received). This is probably due to the salts in the ISO-medium.

Preparation of the test solutions started with loading rates of 1.0, 10 and 100 mg/l applying 1 day of magnetic stirring to reach maximum solubility of the test substance in the test medium. The resulting aqueous mixtures, of which the 1.0 mg/l loading rate was observed to be clear and colourless while the higher loading rates were turbid and increasingly yellow, were filtered through a glass fibre pre-filter (Whatman). The Water Soluble Fractions (WSFs) were used for testing together with a 10% dilution prepared from the 1.0 mg/l WSF.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- 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 5 liters 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 19.3 and 20.0°C
pH:
Between 7.5 and 7.8
Dissolved oxygen:
Between 6.8 and 9.1 mg/l
Nominal and measured concentrations:
Nominal concentration: 0 (control), 10% of a WSF prepared at a loading rate of 1.0 mg/l and WSFs prepared at loading rates of 1.0, 10 and 100 mg/l (concentrations based on the liquid suspension as received).
Measured concentartions: 0 - WSF prepared at a loading rate of 100 mg/l
Based on the TOC content of Biomass residue (liquid) obtained after B2 production (i.e. 13%) the concentration of Biomass residue (liquid) obtained after B2 production in the aquatic samples was calculated and corrected for the control. The initial concentration in the WSF prepared at a loading rate of 100 mg/l was calculated to be 38 mg/l. This concentration decreased to 20 mg/l at the end of the test. Based on these results the average exposure concentration was calculated to be 27 mg/l.
Details on test conditions:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 100 ml, normal headspace
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for the highest concentration, 2 for the lower test 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.
- 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=48 h
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Test concentrations: Control, 10% of a WSF prepared at a loading rate of 1.0 mg/l and WSFs prepared at loading rates of 1.0, 10 and 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.:
27 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Remarks:
Total Organic Carbon converted to test material
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 27 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Remarks:
Total Organic Carbon (TOC) converted to test material
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
100 other: mg/l WSF prepared at 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 (WSF prepared at 100 mg/l)
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
Behavioural abnormalities: no
- Immobility of control: 5% immobility was observed.
- Other adverse effects control: no
Results with reference substance (positive control):
The 24h-EC50 was 0.84 mg/l with a 95% confidence interval between 0.78 and 0.94 mg/l.
The 48h-EC50 was 0.75 mg/l with a 95% confidence interval between 0.56 and 1.0 mg/l.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the present study Biomass residue (liquid) obtained after B2 production did not induce acute immobilisation of Daphnia magna at a loading rate of 100 mg/l (NOEC), corresponding to an average exposure concentration of 27 mg/l.
The 48h-EC50 was above an initial loading rate of 100 mg/l. Based on non-specific analysis the 48h-EC50 was determined to be >27 mg/l.