Registration Dossier

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:
2017-03-06 to 2017-05-08
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD Guideline; EU Method; GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
UVCB

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Duplicate samples from the freshly prepared test medium of the only test concentration and the control were taken at the start of the test.
For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentration during the test period, duplicate samples from the test medium of the only test concentration and the control were collected at the end of the test (after 48 hours).
Additional samples of the control were taken at test start and test end without any sample treatment.

Test solutions

Vehicle:
no
Details on test solutions:
The only test item concentration was nominal 100 mg test item/L. Additionally; a control was tested in parallel (test water without addition of the test item). Thus a limit test was performed in accordance with OECD Guideline 202 to demonstrate that the test item has no toxic effect on the test animals up to this concentration.
Control: In the control, test water was used without addition of the test item.
The test medium of the only test item concentration of nominal 100 mg/L was prepared by dissolving 46.7 mg test item into 467 mL test water by intense stirring for 10 minutes. The test media was prepared just before introduction of the daphnids (= start of the test).

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia magna (Straus), clone 5
Age at Test Start: From 2.0 to 18.75 hours old
Sex: Female
Origin: The daphnids introduced in the test were taken from ibacon's in house laboratory culture.
Breeding Conditions: The daphnids were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in Elendt M4 medium. The test organisms were not first brood progeny. The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of ibacon.

Study design

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

Test conditions

Hardness:
Water Hardness: 2.5 mmol/L (= 250 mg/L) as Calcium Carbonate
Test temperature:
19.7 to 19.8 °C at test start;
19.9 °C at test end
pH:
7.7 to 8.4 at test start;
7.9 to 8.0 at test end; and thus the pH-value did not vary by more than 1.5 units
Dissolved oxygen:
8.8 to 9.2 mg/L at test start;
8.5 to 8.6 mg/L at test end
Nominal and measured concentrations:
100 mg test item/L and a control
Details on test conditions:
Test Environment: Controlled environment room
Measurement of pH, Dissolved
Oxygen and Water Temperature:
The water temperature, pH-values and dissolved oxygen concentrations were determined at test start and test end in each treatment group.
Water Temperature: 19.7 to 19.8 °C at test start;
19.9 °C at test end
pH-Values: 7.7 to 8.4 at test start;
7.9 to 8.0 at test end; and thus the pH-value did not vary by more than 1.5 units
Dissolved Oxygen Concentration: 8.8 to 9.2 mg/L at test start;
8.5 to 8.6 mg/L at test end
Light Regime: 16 h light : 8 h dark
Light Intensity: The light intensity was 280 to 700 lux (measured once during the test).
Reference substance (positive control):
yes
Remarks:
For the evaluation of the quality of the Daphnia clone and the consistency of the experimental conditions, the reference item potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.

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:
NOEC
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
Details on results:
After 48 hours of exposure, no immobilisation of the test animals was observed in the control and in the only test concentration of nominal 100 mg test item/L
Results with reference substance (positive control):
In the most recent test with the reference item potassium dichromate performed in January 2017 the EC50 after 24 hours was determined to be 1.48 mg test item/L, indicating that the sensitivity of the daphnids was consistent with the level proposed by the OECD 202 guideline (EC50-24 h between 0.6 and 2.1 mg potassium dichromate/L).
Reported statistics and error estimates:
No statistical analysis was performed.
The EC50 could not be quantified due to the absence of toxicity of the test item. The NOEC and the LOEC were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
Control Immobilisation Rate was 0 %; no daphnid showed signs of disease or stress. Dissolved Oxygen Concentration was >= 8.5 mg O2/L in the control and test vessels at the end of the test. Thus validity criteria were met.
Conclusions:
The toxic effect of the test item to Daphnia magna was assessed in a static limit test. The 48-hour NOEC was determined to be at least 100 mg test item/L. The NOEC might even be higher than this concentration, but concentrations in excess of nominal 100 mg test item/L have not been tested according to the recommendations of the test guidelines. The 48-hour LOEC and the 48-hour EC50 were clearly higher than 100 mg test item/L. These values could not be quantified due to the absence of toxicity of the test item up to nominal 100 mg test item/L.
The quantification of the test item in the test samples was performed using liquid chromatography with UV detection.
At the start of the test 104% of the nominal test concentration were found. After 48 hours test duration, 104% of the nominal value was determined. During the test the daphnids were exposed to a mean of 104% of nominal. Therefore, all reported results refer to nominal concentrations.
Executive summary:

Title: The test item: Acute Toxicity to Daphnia magna in a Static 48-hour Immobilisation Limit Test

Guidelines: Commission Regulation (EC) No 440/2008, Annex, Part C, C.2.: "Daphnia sp. Acute Immobilisation Test", Official Journal of the European Union (EN), dated May 30, 2008

OECD Guideline for Testing of Chemicals No. 202: "Daphnia sp., Acute Immobilisation Test" adopted April 13, 2004

SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of pre-registration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414

Results

Biological test results: After 48 hours of exposure, no immobilisation of the test animals was observed in the control and in the only test concentration of nominal 100 mg test item/L.

Conclusion: The toxic effect of the test item to Daphnia magna was assessed in a static limit test. The 48-hour NOEC was determined to be at least 100 mg test item/L. The NOEC might even be higher than this concentration, but concentrations in excess of nominal 100 mg test item/L have not been tested according to the recommendations of the test guidelines. The 48-hour LOEC and the 48-hour EC50 were clearly higher than 100 mg test item/L. These values could not be quantified due to the absence of toxicity of the test item up to nominal 100 mg test item/L. The quantification of the test item in the test samples was performed using liquid chromatography with UV detection. At the start of the test 104% of the nominal test concentration were found. After 48 hours test duration, 104% of the nominal value was determined. During the test the daphnids were exposed to a mean of 104% of nominal. Therefore, all reported results refer to nominal concentrations.

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.