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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:
2012
Reliability:
2 (reliable with restrictions)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
Six replicates instead of 4 testing containers have been prepared to have enough solution for chemical analysis. This deviation is not critical nor has any influence on the results of the study.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: white solid
Details on test material:
Batch 2237 JG; expiry date: June 2012

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
The stability of tested concentrations in WAF samples was checked in fresh and old media.

Test solutions

Vehicle:
no
Details on test solutions:
A special solubilisation protocol was carried out before the experimentation according to guidance document OECD 23 on aquatic toxicity testing of difficult substances and mixtures. This method has consisted in stirring three test item nominal concentrations (1, 10 and 100 mg/L) in test dilution water during 24 hours, at obscurity and 20+/-2°C (each concentration was prepared by adding directly the test item in a flask containing test solution). Each test solution plus a dilution water control have been filtered through a membrane filter to remove suspended particles; solutions obtained were named WAF (Water Accommodated Fraction).

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia Magna Straus, clone number 5, from at least the third generation obtained with a non-cyclic parthenogenesis reproduction.
The daphnids breeding is carried out in closed bottles placed in a climatic chamber free from any toxic vapor. Daphnids used in this experimentation were previously filtered in order to obtain animals of less than 24 hours old.

Study design

Test type:
semi-static
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
None

Test conditions

Hardness:
260 mg/L of CaCO3
Test temperature:
19.8-20.3°C
pH:
8.0-8.3
Dissolved oxygen:
8.6-9.1 mg/L
Nominal and measured concentrations:
Nominal concentration: 1 mg/L test item
Measured concentration at T0: < 0.15 mg/L APG
Measured concentration at T24h (before renewal) - biotic samples: < 0.15 mg/L APG (below LOQ)
Measured concentration at T24h (before renewal) - abiotic samples: < 0.15 mg/L APG (below LOQ)

Nominal concentration: 10 mg/L test item
Measured concentration at T0: 0.81 mg/L APG
Measured concentration at T24h (before renewal) - biotic samples: 0.61 mg/L APG (-25%)
Measured concentration at T24h (before renewal) - abiotic samples: 0.97 mg/L APG (+20%)

Nominal concentration: 100 mg/L test item
Measured concentration at T0: 8.26 mg/L APG
Measured concentration at T24h (before renewal) - biotic samples: 7.31 mg/L APG (-12%)
Measured concentration at T24h (before renewal) - abiotic samples: 9.40 mg/L APG (+14%)
Details on test conditions:
The test contained a total of 30 glass test tubes as follows:
- unfiltered control: 6 replicates
- Filtered control: 6 replicates
- Concentration 1 mg/L: 6 replicates
- Concentration 10 mg/L: 6 replicates
- Concentration 100 mg/L: 6 replicates
Each glass test tube contained 10 mL of test solution and 5 daphnids; they have been covered in oreder to avoid evaprtation and dust deposition.

Incubation:
Test test was performed in the dark at 20+/-1°C. Daphnids were not fed during the test period.
Observations of the daphnid's mobility were made after 24 and 48 hours of exposition to the test item, with renewal of test solutions at 24h .
Every sign of stress observed on the daphnids after 48 hours of exposition to the test item have been reported. pH and dissolved oxygen have been measured at the beginning and at the end of the test for each test solutions (including both control).
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.
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Details on results:
In all samples, after 24h and 48h, no immobilised daphnids were observed.
No sign of stress was observed on the mobile daphnids after 48 hours of exposition.
Results with reference substance (positive control):
The last toxic response obtained on the reference item K2Cr2O7 gave an EC50-24h of 1.12 mg/L (1.01-1.24 mg/L), 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 May 1996 as mentioned in the OECD guideline) and in accordance with historical data obtained by the laboratory.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In the conditions of this semi-static test (WAF approach), the EC50-48 hours of the test item "2237 JG" on Daphnia Magna was superior to 100 mg 2237 JG/L, value expressed in nominal concentration.
Executive summary:

In a semi-static acute toxicity test performed according to OECD guideline 202, daphnids (Daphnia magna) were exposed for 48 hours to three nominal concentrations ranging between 1 and 100 mg 2237 JG/L water.

Since the test item was insoluble in the tested medium, the use of Water Accommodated Fractions (WAF) for the preparation of the test media was necessary.

The test item stability was checked by the determination of Alkyl Poly Glucosid (APG) in fresh and old WAF (GC-FID; non GLP data). Actual concentrations of APG were in good agreement with the initial concentrations except for the 10 mg/L test concentration.

After 24 and 48 hours of exposure, no immobilisation was observed atthe three test concentrations.

 

EC50(24 h) > 100 mg/L (nominal)

EC50(48 h) > 100 mg/L (nominal)