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:
2010
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
The concentration of methanol in the solvent control was higher than demanded in the guideline. As no significant immobilisation occurred during the test, this deviation was stated as uncritical.
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
yes
Remarks:
The concentration of methanol in the solvent control was higher than demanded in the guideline. As no significant immobilisation occurred during the test, this deviation was stated as uncritical.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
After 0, 24 and 48 hours 1 ml test solution (seven replicates inflow and outlet of the treatment and two replicates solvent control) were taken and transferred into an HPLC vial. To each sample; 1 % formic acid (10µl/ml) was added. Then the samples were frozen immediately. Each sample was labelled with the project number of the study, the nominal concentration, the sampling time and the replicate number.
Five replicates of test solution sample and one replicate of solvent control sample were shipped to the test site Henkel AG & Co. KGaA. The other replicates remain in the test facility as retain sample until the end of the analytical phase at -20°C.

Test solutions

Vehicle:
yes
Details on test solutions:
A stock solution containing 100 g/L in Methanol was prepared. For the solvent control, methanol in a concentration of 2 ml/L was used.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia magna
Variety: STRAUS
Strain: Berlin
Sex: female
Age: between 0 and 24 hours
Origin: Umweltbundesamt Berlin
Arrival of strain: 27. September 2007

Study design

Test type:
flow-through
Water media type:
saltwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Test temperature:
19.7 - 20.8°C
pH:
7.6 - 8.4
Dissolved oxygen:
9.3 - 9.7 mg/L
Nominal and measured concentrations:
The measured concentration after 24 and 48 hours was in a range of 103 % and 128 % of the nominal concentration. The samples of the first day arrived at the laboratory of the test site with a delay of several hours, although shipment had been made on time. Those test solutions were no longer frozen and degradation of the test item occurred. Therefore, the measured concentration at the start was lower than the initial concentration. The determination of the biological results was based on the mean of the measured concentrations for the samples after 24 and 48 hours and the nominal concentration at the beginning.
Details on test conditions:
Date of performance: 22. - 24. March 2010
Type of test: flow through
Treatments: 100 mg/L
Stock solution: 100 g/L (in methanol)
Duration: 48 hours
Observation times: 24 and 48 hours
Vessels: glass aquaria with outlet pipe, nominal volume 2800 mL (volume including to grille boxes: 2490 mL)
Feeding: none
Lighting: none
Replicates: one vessel, containing test solution and 2 grille boxes with 10 daphnia each
Solvent Control: one vessel, containing dilution water and 2 grille boxes with 10 daphnia each
Control: one vessel, containing dilution water and 2 grille boxes with 10 daphnia each
Flow Rate Stock Solution: 25 µl/min (Stock solutions were freshly prepared daily)
Flow Rate Dilution Water: 25 mL/min
Medium Reneval: 1 / 99.5 min

Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
106 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
106 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 106 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 106 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Results with reference substance (positive control):
NOEC 24 h 0.5 mg/L
24 h EC50i 1,2 mg/L
24h EC100i 2,5 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The determination of the biological results was based on the mean of the measured concentrations for the samples after 24 and 48 hours (a range of 103 - 128 mg/L) and the nominal concentration at the start of the test.
The following results were determined for the test item B 1061 (species. Daphnia magna):
24h-NOEC = 106 mg/L
48h-NOEC = 106 mg/L
24h-EC50i > 106 mg/L
48h-EC50i > 106 mg/L
Executive summary:

As the test item B 1061 wasn't stable in aqueous solution, the main study was performed under flow-through conditions. In a non-GLP pre-test under static conditions, toxicity was observed. In a non-GLP range finding test under flow-through conditions, no toxicity was observable. Therefore, it is believed that toxicity was caused by metabolites of the test item. The main study (flow-through test) was performed as a limit test using a single concentration (100 mg/L).

Twenty daphnia were exposed to the test item for 48 hours in a flow-through system. After 24 and 48 hours, the immobilised daphnia were counted.

In the treatment, only 10% immobilisation was observed. None of the animals were immobilised in the control, 5% were immobilised in the solvent control. The immobilisation rates can be stated as not significant.

The 24h-EC50i of potassium dichromate was tested in a current reference test. The value was determined as 1,2 mg/L, lying within the demanded range of 0.6 - 1.7 mg/L.

After the start, after 24 and after 48 hours the content of the test item in the inflow and the outlet of the aquaria were determined using APCI-MS (Atmospheric Pressure Chemical Ionisation-Mass Spectrometer).

The applied concentration of B 1061 in the Daphnia magna solutions (100 mg/L) could be confirmed. The measured concentrations after 24 and 48 hours were in the range of 103 - 128 mg/L. Concerning the first day of sampling (22.03.2010) it has to be considered that the samples arrived at the laboratory of the test site with a delay of several hours, although leaving the test facility at the same time as on the following days. Those test solutions were no longer frozen, and degradation of the test item occurred. Therefore, the measured concentration at the start was longer than the initial concentration in the test solution at the beginning of the study.

The determination of the biological results was based on the mean of the measured concentrations for the samples after 24 and 48 hours and the nominal concentration at the start of the test.

The following results were determined for the test item B 1061 (species. Daphnia magna):

24h-NOEC = 106 mg/L

48h-NOEC = 106 mg/L

24h-EC50i > 106 mg/L

48h-EC50i > 106 mg/l