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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
Study period:
September 2017
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:
yes
Remarks:
It concerns the number of replicates for the concentration 100 mg/L: 8 replicates instead of 4 r eplicates were done to have enough solution at the end of the test for the chemical analyses. This has no influence on the results of this study
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 (September 21, 2017) indicates an EC50-24h of 1.34 mg/L (1.22 – 1.47), 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 test item 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 explained in the attached READ-ACROSS Argumentation, it can be concluded that the results of the study made on Daphnia magna, acute immobilization test performed on the reference substance “Polysaccharides of glucose from Oat” is likely to predict the properties of the target substance “Oligosaccharides of Caesalpinia Spinosa extract” 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:
2017
Report Date:
2017

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: beige powder
- Storage: in a dark place at room temperature
- Batch : 14 262
- Manufacturing date: September 19th 2014
- Expiry date: September 19th 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.364-e version 2: Method for the chemical analysis of Total Carbon (TC) in samples containing the test item from daphnids ecotoxicology solutions by a Total Organic Carbon Analyzer (described in the study 16/1252F/e and the following protocol:
- In Fresh solutions (at the beginning of the test:T0h and for the renewal at T24h), these solutions were 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 were pooled and analyzed
- for the Control (i.e. daphnids test medium filtered), and the concentration 100 mg/L
These solutions were analysed taken directly from the starting solutions without daphnids.
The measured concentrations of the test item should not vary more than ± 20 % during the test (48h).

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:
19.8 – 20.0°C
pH:
pH (Dilution water M4) = 8.2
Dissolved oxygen:
9.0 mg/L
Salinity:
no data
Conductivity:
-
Nominal and measured concentrations:
Nominal concentration: 100 mg/L
Measured concentration: 101.3 mg/L (at initial time) and 99.5 mg/L (after 48 hours of exposure)
Details on test conditions:
TEST SYSTEM
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).
This study was conducted on one concentration levels: 100 mg/L.
The test contained a minimum of 12 glass test tubes as follow:
• control: 6 replicates
• concentration 100.0 mg/L: 6 replicates
Each glass test tube (sterilized by autoclave) contained 10 mL of test solution and 5 daphnids; they were covered in order to avoid evaporation and dust deposition
OTHER TEST CONDITIONS
- Light intensity: Darkness
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Observations of the daphnid’s mobility were done after 24 and 48 hours of exposition to the test item.
Every sign of stress observed on the daphnids after 48 hours of exposition to the test item were reported if need be.
pH and dissolved oxygen were measured at the beginning and at the end of the test for each test solutions (including the controls).
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 (September 21, 2017) indicates an EC50-24h of 1.34 mg/L (1.22 – 1.47), 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 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 on the daphnids Daphnia magna, according to the OECD 202 guideline, at 100 mg/L, limit test, in static conditions.

The concentrations which immobilise 50% of the daphnids exposed to the test item after 24 and 48hours, 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 is considered as stable in the testing conditions because the measured concentrations of the did not vary more than ±20 % of initial measured concentration during the test.

The results obtained for the test item on the toxicological endpoint “immobilisation” give an EC50-24h and anEC50-48h superior to the highest tested concentration,i.e. 100mg/L,value expressed in nominal test item concentration.