Registration Dossier

Diss Factsheets

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:
2018-12-03 - 2018-12-06 (experimental phase)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2019

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
adopted 13. Apr. 2004: ”Daphnia sp., Acute Immobilisation Test“
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
adopted 30. May 2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: OECD guidance document no. 23
Version / remarks:
second edition, GUIDANCE DOCUMENT ON AQUATIC TOXICITY TESTING OF DIFFICULT SUBSTANCES AND MIXTURES, adopted 14. Dec. 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Test material form:
solid: particulate/powder

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 4.6 / 10 / 22 / 46 / 100 mg/L
- Sampling method: A saturated solution was prepared for the test. This was done by mixing the nominal load of 100 mg/L (real load: 100.8 mg/L) with the corresponding amount of dilution water and shaking vigorously for 23.75 hours. The resulting solution was filtrated through 0.45 µm PTFE filters.
The lower treatments (4.6 / 10 / 22 / 46 mg/L) were prepared by dilution of this saturated solution with dilution water.

- Sample storage conditions before analysis: n.a.

Test solutions

Vehicle:
no
Remarks:
dilution water was used
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A saturated solution was prepared for the test. This was done by mixing the nominal load of 100 mg/L (real load: 100.8 mg/L) with the corresponding amount of dilution water and shaking vigorously for 23.75 hours. The resulting solution was filtrated through 0.45 µm PTFE filters.
The lower treatments (4.6 / 10 / 22 / 46 mg/L) were prepared by dilution of this saturated solution with dilution water.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain/clone: Daphnia magna STRAUS Berlin
- Justification for species other than prescribed by test guideline: n.a.
- Age at study initiation (mean and range, SD): between 0 and 24 hours
- Source: Umweltbundesamt Berlin
In-house breeding since: 27. September 2007
Daphnia magna is bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. They multiply by parthenogenesis, thus being genetically identical. The keeping is performed similar to the method described in the OECD guideline.
- Feeding during test : no
Vessels preserving glasses, nominal volume 2 L
Medium M4-Medium (recipe of ELENDT)
Food green algae (Desmodesmus subspicatus)
Medium renewal twice a week
Photo period 16/8 hours, using neon tubes
Temperature 20 +- 2 °C

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS:
Selection of Daphnia
18.25 hours before the start of the test, the adult animals were separated from the young. 1 hour before test start, the adults were caught with the help of a glass tube, and the new-born daphnia, aged between 0 and 17.25 hours, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.
Switching from M4-medium (husbandry) to Dilution water (test) has been shown not to cause any detrimental effects for test daphnia.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
none

Test conditions

Hardness:
Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250
Test temperature:
19.6 – 21.3 °C
pH:
7.7 - 7.9
Dissolved oxygen:
8.6 - 8.8 mg/L
Nominal and measured concentrations:
4.6 / 10 / 22 / 46 / 100 mg/L (nominal); the concentrations to be tested are based on the result of a non-GLP pre-test.
Because of the very low solubility of the test item the measured concentrations at the start and the end of the test lay in the range of the blank control. Therefore, the nominal concentrations were used for the evaluation of the biological results.
Details on test conditions:
TEST SYSTEM
- Test vessel: glass beakers, nominal volume 50 mL, tall shape
- Volume of solution: 20 ± 5 mL
- Aeration: After preparation, the dilution water was aerated.
- Renewal rate of test solution (frequency/flow rate): none
- 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 (deviations from the nominal weighted loads were less than 5 %):
293.8 mg/L CaCl2*2H2O
123.3 mg/L MgSO4*7H2O
64.8 mg/L NaHCO3
5.8 mg/L KCl

OTHER TEST CONDITIONS
- Adjustment of pH: no. After preparation, the pH was measured (7.8).
- Photoperiod: 16/8 light/dark
- Light intensity: neon tubes

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
After 24 and 48 hours, the immobilised daphnia were counted.
The pH, the concentration of dissolved oxygen and the content of the test item in the test vessels were measured at the beginning and at the end of the test.

VEHICLE CONTROL PERFORMED: n.a.

RANGE-FINDING STUDY
- Test concentrations / Results used to determine the conditions for the definitive study: The concentration to be tested was based on the result of a non-GLP pre-test.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Limit test: no
- Dose-response test: yes
- ECx: The 24h-EC50i value was determined as 2.05 mg/L. The value was within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of potassium dichromate K2Cr2O7 as specified in the OECD guideline.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The study was conducted under GLP according to OECD 202. The available information allows the conclusion that the test was properly conducted, all criteria for acceptability of the test were met, this study was considered to be valid. The following results were determined for the test item Polyperin Y (species: Daphnia magna):

48h-NOEC ≥ 100 mg/L
48h-LOEC > 100 mg/L
48h-EC50 > 100 mg/L

Classification:
- Acute hazard: No acute aquatic toxicity recorded at levels up to the limit of water solubility;
- Long-term hazard: No adequate chronic toxicity data available for all three trophic levels.
Substance is nevertheless of concern based on the following findings:
- Poorly soluble substance (WS < 1 mg/L);
- No acute aquatic toxicity recorded at levels up to the limit of water solubility;
- Not rapidly degradable;
- High potential for bioaccumulation (in absence of BCF data, log Kow > 4);
- No evidence on NOEC being > water solubility for all three trophic levels;
- No other evidence of rapid degradation in the environment.
-> "safety net" classification as aquatic chronic 4, H413
Executive summary:

Determination of short-term toxicity of Polyperin Y against Daphnia magna STRAUS according to OECD 202 resp. EU C.2 (GLP):

One valid experiment was performed. The study was performed using 5 concentrations ranging from 4.6 to 100 mg/L (nominal). For each test concentration and the blank control, 20 Daphnia were exposed to the test item for 48 hours in a static test system. After 24 and 48 hours, the immobilised Daphnia were counted. None of the animals was immobilised in the blank control and the concentrations.

Potassium dichromate K2Cr2O7 (CAS 7778-50-9) was used as positive control in a current reference study to assure that the test conditions are reliable.

At the start and at the end of the test, the content of the test item in the test solutions was estimated by determination of the dissolved organic carbon (DOC) content in the test solutions using a carbon analyser.

Because of the very low solubility of the test item the measured concentrations at the start and the end of the test lay in the range of the blank control. Therefore, the nominal concentrations were used for the evaluation of the biological results.

At the acute toxicity test against Desmodesmus subspicatus, significant inhibition of algal growth was observed at the highest tested concentration 100 mg/L. Therefore, the test item must have been present in the test solution, but wasn´t measurable. The dilution water, which was used in this study, is comparable to the algal medium, which was used in the acute toxicity test against Desmodesmus subspicatus. Therefore, it can be assumed that test item was present in the test solutions in this study, but the test system was not sensitive enough to show toxicity. The following results were determined for the test item Polyperin Y (species: Daphnia magna):

48h-NOEC ≥ 100 mg/L

48h-LOEC > 100 mg/L

48h-EC50 > 100 mg/L