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:
January 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

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

Test material

Constituent 1
Reference substance name:
Almond, sweet, ext.
EC Number:
291-063-5
EC Name:
Almond, sweet, ext.
Cas Number:
90320-37-9
IUPAC Name:
Proteins from sweet almond extract
Test material form:
solid: particulate/powder
Details on test material:
- Appearance: white powder
- Storage: in a dark place at room temperature
- Batch : 16 187
- Manufacturing date: July 5th 2016
- Expiry date: July 5th 2018

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 ecotoxicological testing, chemical analyses in daphnids test solutions were performed according to the Analytical Protocol FCBA Chimie No.345-e version 2: Method for the chemical analysis of Total Carbon (TC) in samples containing the test item POLYLIFT AT from daphnids ecotoxicology solutions by Carbon Analyzer (described in the study 16/1182F/a) and the following protocol:
- In Fresh solutions (at the beginning of the test:T0h and for the renewal at T24h), these solutions will be analysed taken directly from the starting solutions without daphnids
- in Old solutions (before renewal T24h and at the end of the test T48h), two test tubes will be pooled and analyzed
- for the Control (i.e. daphnids test medium filtered), and the concentration 100 mg/L

Schedule analysis at initial time:
Control Fresh T0h 1 replicate
100 mg/L Fresh T0h 2 replicates

These solutions will be analysed taken directly from the starting solutions without daphnids.

Schedule analysis at T24h:
Control Old T24h 1 replicate
100 mg/L Old T24h 2 replicates
Control Fresh T24h 1 replicate
100 mg/L Fresh T24h 2 replicates


Schedule analysis at final time T48h:

Control Old T48h 1 replicate
100 mg/L Old T48h 2 replicates

Test solutions

Vehicle:
no
Details on test solutions:
The test item was soluble in the test medium.
At the request of the sponsor, 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 µm 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:
Daphnia magna Straus, clone number 5, from at least the third generation (i.e. the third brood) obtained with a non-cyclic parthenogenesis reproduction.
The daphnids breeding was carried out in closed bottles placed in a climatic chamber free from any toxic vapour and in medium M4.
Daphnids used in this experimentation were previously filtered in order to obtain animals from a size between 560 µm and 800 µm (corresponding to daphnids less than 24 hours old).

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
none

Test conditions

Hardness:
260 mg/L of CaCO3
Test temperature:
19.9 – 20.1°C
pH:
pH (Dilution water M4) = 7.4
Dissolved oxygen:
9.1 mg/L
Salinity:
no data
Nominal and measured concentrations:
Nominal concentration: 100 mg/L
Measured concentration: 106.6 mg/L (at initial time) and 115.8 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
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 (January 16, 2017) gives an EC50-24h of 1.16 mg/L (1.02 – 1.33), 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 “POLYLIFT AT” 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:

The aim of this study was to assess the effects of the test item “POLYLIFT AT”on the daphnids Daphnia magna, according to the OECD 202 guideline, at 100 mg/L, limit test, in semi-static conditions.

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.

At the nominal concentration of 100 mg/L, the test item « POLYLIFT AT» is considered as stable in the testing conditions because the measured concentrations of the “POLYLIFT AT” did not vary more than ± 20 % of initial measured concentration during the test.

The results obtained for the test item “POLYLIFT AT” on the toxicological endpoint “immobilisation” give an EC50-24h and anEC50-48h higher than the highest tested concentration,i.e. 100 mg/L, based on nominal test item concentration.