Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
July 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose:
read-across source
Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
other information
Justification for type of information:
Cf the attached Read across argumentation
Reason / purpose:
read-across: supporting information
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: highest tested concentration
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: highest tested concentration
Details on results:
- No immobilization was observed at 24 h and 48 h (including controls).
- No sign of stress was observed on the mobile daphnids after 48 h of exposure
Results with reference substance (positive control):
The last toxic response obtained on the reference item K2Cr2O7 (March 24, 2016) gives an EC50-24h of 1.58 mg/L (1.44 – 1.76), indicating that the daphnids sensitivity is correct (within the accepted interval of 0.6–2.1 mg/L fixed by the International Standard ISO 6341 December 2012 as mentioned in the OECD 202 Guideline) and in accordance with historical data obtained by the laboratory
Validity criteria fulfilled:
yes
Conclusions:
The results obtained for the reference substance on the toxicological endpoint “immobilisation” give an EC50-24h and an EC50-48h higher than the highest tested concentration, i.e. 100 mg/L, based on nominal test item concentration
Executive summary:

Based on the consideration above, it can be concluded that the results of the study made on Daphnia magna, acute immobilization test performed on the reference substance Proteins hydrolyzates of Lupin (Lupinus Albus)” is likely to predict the properties of the target substance “Proteins hydrolyzates of Rice (Oryza Sativa)” and are considered as adequate to fulfil the information requirement of the Annex VII 9.1.1: 1. Short term toxicity to aquatic invertebrates.

=> EC50-24h and EC50-48h > 100 mg/L (highest tested concentration)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Appearance: Yellow powder
- Storage In darkness at room temperature
- Batch: 15 288
- Manufacturing date: October 2015
- Expiry date: October 2017

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
In order to verify the initial concentrations and maintenance of the exposure concentrations of the test item during the exposure period, chemical analysis of the "ANAGELINE AT" in daphnids test solutions were performed at the beginning and after 24 h (before renewal). Method for the chemical analysis of Total Carbon (TC) in samples containing the test item ANAGELINE AT from daphnids ecotoxicology solutions by Total Organic Carbon Analyzer (described in the study 15/1256F/a) and the following protocol:
- in fresh solutions at the beginning of the test (T=0h)
- in old solutions (T=24h)
- for the control (i.e. daphnids test medium filtered) and concentration 100 mg/L

Schedule analysis at initial time:
-Filtered control Abiotic: 1 replicate
-100 mg/L Abiotic: 2 replicates
These solutions were analysed taken directly from the starting solutions without daphnids.
Schedule analysis at final time (before renewal: 24h):
-Filtered control Biotic: 1 replicate
-100 mg/L Biotic: 2 replicates

Test solutions

Vehicle:
no
Details on test solutions:
The test item is soluble and directly dispersed in test medium. The test medium and the solution (100 mg test item/L) were filtered through a membrane filter (cellulose acetate) with a mesh size of 0.22 um to sterilizing and all the manipulations were done under sterile conditions to prevent contamination.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Daphnia magna Straus, clone number 5
- Source: from at least the third generation (i.e. the third brood) obtained with a non-cyclic parthogenesis reproduction.
- Age at study initiation: less than 24 hours old.
- Method of bredding: the daphnids breeding was carried out in closed bottles placed in a climatic cha
mber free from any toxic vapour and in medium M4.
- Feeding during test: None

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Remarks on exposure duration:
None

Test conditions

Hardness:
250 mg/L of CaCO3
Test temperature:
19.9 - 20.1°C
pH:
8.1
Dissolved oxygen:
8.8 mg/L
Salinity:
No data
Conductivity:
No data
Nominal and measured concentrations:
Nominal concentration: 100 mg/L
Measured concentration: 101.91 mg/L (at initial time) and 97.7 mg/L (after 24 hours of exposure)
Details on test conditions:
TEST SYSTEM
- Test vessel: glass test tube
- Type: closed
- Each glass test tube (sterilized by autoclave) contained 10 mL of test solution and five daphnids; they were covered in order to avoid evaporation and dust deposition.
- Renewal rate of test solution: Renewal of test solutions at 24h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6

OTHER TEST CONDITIONS
- Light intensity: Darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Observations of the daphnid's immobility and behaviour were carried out after 24 and 48 h in control and exposed groups.
- Any sign of stress observed on the daphnids after 48 hours of exposure to the test item were reported.
- pH and dissolved oxygen were measured before and after renewal of solutions (including the controls).

RANGE-FINDING STUDY
- Test concentrations: 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: No immobilization was observed at any of the concentration tested. Based on the results of the preliminary test, it has been proposed to perform a limit test at 100 mg/L for the study. This study was conducted on one concentration levels: 100 mg/L.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
Details on results
- No immobilization was observed at 24 h and 48 h (including controls).
- No sign of stress was observed on the mobile daphnids after 48 h of exposure
Results with reference substance (positive control):
The last toxic response obtained on the reference item K2Cr2O7 (June 27, 2016) gives an EC50-24h of 1.24 mg/L (1.13 - 1.37), indicating that the daphnids sensitivity is correct (within the accepted interval of 0.6 - 2.1 mg/L fixed by the International Standard ISO 6341 December 2012 as mentioned in the OECD 202 Guideline) and in accordance with historical data obtained by the laboratory.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The results obtained for the test item "ANAGELINE AT" on the toxicological endpoint "immobilisation" give an EC50-24h and an EC50-48h > 100 mg/L, based on nominal test item concentration.
Executive summary:

The aim of this study was to assess the acute toxicity of the test item "ANAGELINE AT" on the freshwater invertebrate Daphnia magna, according to the OECD 202 guideline. The study was performed at 100 mg/L, limit test, under semi-static conditions, with GLP compliance.

The test was performed in sterilized vessels and sterile conditions with renewal at 24 h of the test solution. Groups of thirty Daphnia magna, less than 24 h old, were exposed for 48h to a control solution (Elendt M4 medium) and to the test item “ANAGELINE AT”, prepared in Elendt M4 medium.

The test item concentration was prepared under sterile conditions in order to obtain the required nominal concentration of 100 mg/L Elendt M4 medium.

The concentrations which immobilise 50% of the daphnids exposed to the test item after 24 and 48 hours, reported as EC50 -24h and EC50 -48h were evaluated, relative to the tested concentration: 100 mg/L. After 48 h, no immobilization was observed in any test tubes.

In order to verify the initial concentrations and maintenance of the exposure concentrations of the test item during the ecotoxicological testing, a chemical analysis of "ANAGELINE AT" in the test solutions was performed. At the nominal concentration of 100 mg/L, the test item "ANAGELINE AT" is considered as stable in the testing conditions because the measured concentrations of the "ANAGELINE AT" did not vary more than +/- 20% of initial measured concentration during the test.

In the control, not more than 10 % of the daphnids showed an immobilization. In addition, the dissolved oxygen concentration was determined to be higher than 3 mg/L in the control and test vessels. Thus, the validity criteria were respected.

The results obtained for the test item "ANAGELINE AT" on the toxicological endpoint "immobilisation" give an EC50-24h and an EC50-48h > 100 mg/L, based on nominal test item concentration.