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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Purity: 28 wt%

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: One sample plus a back-up sample from each test concentration, including the dilution water control, on day 0 of the test before the test solutions were poured into the replicate test chambers. In addition to the neat test solution samples, one sample plus a back-up sample diluted 4x into methanol (5 mL test solution + 15 mL methanol) from each test concentration, including the dilution water control, on day 0 of the test before the test solutions were poured into the replicate test chambers. One sample plus a back-up sample from 2 of the 4 replicate chambers (replicates B and D) at each test concentration, including the dilution water control, at test end (day 2). In addition to the neat test solution samples, one sample plus a back-up sample diluted 4x into methanol (5 mL test solution + 15 mL methanol) from the 2 replicate chambers (replicate B and D) at each test concentration, including the dilution water control, at test end.
- Sample storage conditions before analysis: All samples and back-up samples were stored refrigerated when not in use.

Test solutions

Vehicle:
no
Details on test solutions:
Test Solution Preparation
Test substance solutions were prepared by dilution from a stock solution of test substance in the testing laboratory’s well water. A 712 mg/L stock solution was prepared by adding approximately 1.42 g of test material to well water in a 2-L glass beaker (final volume 2L), sonicating for approximately 35 minutes, and stirring for approximately 30 minutes. The stock solution was also the highest test concentration. Test solutions were prepared by adding the appropriate volume of the stock solution to the appropriate volume of well water in 1-L glass beakers (final volume 1L) and stirring for approximately 15 minutes. The stock solution and all test solutions were clear and colourless with a foamy surface but no visible precipitate.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia magna were reared at the testing laboratory in 250-mL (1 per beaker) or 1000-mL Pyrex® beakers (10 per beaker at culture initiation) which contained 200 mL or 1000 mL of aerated, filtered testing laboratory well water held at approximately 20°C. Daphnids were fed on a daily basis with 3 mL/L of a yeast, cereal leaves and trout chow (YCT) mixture (standardized to 1700 to 2100 mg/L total solids) and the green alga, Pseudokirchneriella subcapitata, at a rate of approximately 62,500 cells/mL of culture medium. The combination of YCT and alga is equivalent to approximately 0.1-0.2 mg total organic carbon per daphnid. Neonates used in this test were less than 24 hours old and were collected from the 7th brood of 21-day old parent daphnids. Sickness, injury, and abnormalities were not seen and ephippia were not being produced by the parent daphnids. No adult immobility was seen in the cultures used for testing during the 48-hour pretest period. Daphnia magna were identified by labels on the culture beakers and test chambers. Daphnids were not fed during the test.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
164 to 178 mg/L as CaCO3
Test temperature:
20.9°C (mean) with a range of 20.1 to 21.6°C
pH:
pH
0 Hours: 8.1 – 8.2
48 Hours: 8.3
Dissolved oxygen:
DISOLVED OXYGEN
0 Hours: 8.0-8.6 mg/L
48 Hours: 8.5-8.7 mg/L
Nominal and measured concentrations:
Nominal concentrations: 0 (control), 44.5, 89, 178, 356, and 712 mg/L
Nominal total fluorinated solids concentrations: 0 (control), 13.1, 26.2, 52.3, 105, and 209 mg/L
Mean, measured total fluorinated solids concentrations: 0 (control), 9.18, 16.7, 33.9, 97.3, and 122 mg/L

Total fluorinated solids includes 28.0% amphoteric fluorinated surfactant, 1.4% impurity 1, and < 0.2% impurity 2.
Mean, measured total fluorinated solids concentration was calculated as (Day 0 centrifuged + Mean Day 2 centrifuged)/2.
Mean measured concentrations ranged from 58 to 70% of the nominal concentrations adjusted for 29.4% solids by analysis.
The level of fluorinated solids in the dilution water control was below the limit of quantitation of 1.68 mg/L after application of a 160x dilution factor.
Details on test conditions:
TEST SYSTEM
- Test vessel: glass beakers (250-mL) containing approximately 200 mL of test solution (approximately 6.5-cm test solution depth); covered with a glass plate during the test.
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS:
- Source/preparation of dilution water: Test laboratory well water adjusted to approximately 100-140 mg/L as CaCO3 by the flow-proportioned addition of CaCl2. Then aerated, passed through a green sand filter to remove iron, and filtered through 50-, 10-, and 0.45-µm filters to remove particulates.
- Conductivity: 270 to 292 µmhos/cm

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light / 8 hours darkness, included 30 minutes of transitional light preceding and following the 16-hour light interval.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobility and sublethal effects (once daily, 24 and 48 hours).

RANGE-FINDING STUDY
A range-finding study with 1 test chamber each containing 10 organisms per test substance concentration was conducted using a dilution water control and nominal test substance concentrations of 0.12, 1.2, 12, and 120 mg total fluorinated solids/L. At the end of 48 hours, the respective immobility values were 0, 0, 0, 0, and 50%. All test substance solutions except the nominal 120 mg total fluorinated solids/L concentration were clear and colourless with no visible precipitate at test start. The nominal 120 mg total fluorinated solids/L test substance concentration was clear and colourless with film and bubbles on the surface at test start.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 712 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobility
Remarks on result:
other: highest concentration tested
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 122 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
other: total fluorinated solids
Basis for effect:
other: immobility
Remarks on result:
other: highest concentration tested
Details on results:
No immobility was observed in the dilution water control daphnids. Daphnids (12 and 7) were observed floating at the surface at 24 and 48 hours, respectively, in the dilution water control. Daphnids (2 and 1) were observed floating at the surface at 24 and 48 hours, respectively, in the mean, measured 33.9 mg total fluorinated solids/L concentration. One daphnid was observed floating at the surface at 24 hours in the mean, measured 67.3 mg total fluorinated solids/L concentration. Surviving daphnids were exhibiting erratic swimming and lethargy at 24 and 48 hours in the mean, measured 122 mg total fluorinated solids/L concentration. The highest nominal concentration causing no immobility at test end was 356 mg/L (mean, measured 67.3 mg total fluorinated solids/L). The lowest nominal concentration causing 100% immobility at test end was greater than 712 mg/L (mean, measured 122 mg total fluorinated solids/L). Nominal concentrations of the test substance and mean, measured concentrations of the total fluorinated solids in the test substance were used for calculation of EC50 values. The 48-hour EC50, based on nominal and mean, measured concentrations and immobility, was greater than 712 mg/L (122 mg total fluorinated solids/L).
Reported statistics and error estimates:
The 48-hour EC50 values were not calculated because test organism immobility did not exceed 50% in any test substance concentration.

Applicant's summary and conclusion

Conclusions:
This study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).
48-hour EC50, based on nominal concentrations and immobility > 712 mg/L.
48-hour EC50, based on mean, measured concentrations and immobility >122 mg total fluorinated solids/L
Executive summary:

The acute toxicity of the test substance to unfed Daphnia magna neonates, less than 24 hours old at test start, was determined in an unaerated, 48-hour, static test. The study was conducted with 5 concentrations and a dilution water control at a mean temperature of 20.9°C (range of 20.1-21.6°C). Four replicates with 5 daphnids per replicate were used per test substance concentration and dilution water control. The test substance contained 29.4% total fluorinated solids (total fluorinated solids includes 28.0% amphoteric fluorinated surfactant, 1.4% impurity 1, and < 0.2 % impurity 2) by analysis. Exposure of daphnids to nominal concentrations of 44.5, 89, 178, 356, and 712 mg/L (which correspond to nominal total fluorinated solids concentrations of 13.1, 26.2, 52.3, 105, and 209 mg total fluorinated solids/L) resulted in 0, 5, 0, 0, and 35% immobility, respectively, at the end of 48 hours. Mean, measured total fluorinated concentrations ranged from 58 to 70% of the nominal concentrations. No immobility was observed in the dilution water control daphnids. Daphnids (12 and 7) were observed floating at the surface at 24 and 48 hours, respectively, in the dilution water control. Daphnids (2 and 1) were observed floating at the surface at 24 and 48 hours, respectively, in the mean, measured 33.9 mg total fluorinated solids/L concentration. One daphnid was observed floating at the surface at 24 hours in the mean, measured 67.3 mg total fluorinated solids/L concentration. Surviving daphnids were exhibiting erratic swimming and lethargy at 24 and 48 hours in the mean, measured 122 mg total fluorinated solids/L concentration. The highest nominal concentration causing no immobility at test end was 356 mg/L (mean, measured 67.3 mg total fluorinated solids/L). The lowest nominal concentration causing 100% immobility at test end was greater than 712 mg/L (mean, measured 122 mg total fluorinated solids/L). Nominal concentrations and mean, measured concentrations of the total fluorinated solids in the test substance were used for calculation of EC50 values. The 48-hour EC50, based on nominal and mean, measured concentrations and immobility, was greater than 712 mg/L (122 mg total fluorinated solids/L).