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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:
19 Sep - 06 Oct 2016
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)
Version / remarks:
13 Apr 2004
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Landesanstalt für Umwelt, Messungen und Naturschutz Baden-Württemberg, Germany (10 Dec 2015)

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs.
EC Number:
614-257-7
Cas Number:
68071-40-9
Molecular formula:
C18H34O4, C20H38O4 and C22H42O4 (mainly)
IUPAC Name:
Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: Control and limit concentration of 10.0 mg/L (nominal)
- Sampling method: Analytical samples were taken from the test item loading rates and control at test start (0 h), after 24 h from fresh and aged solutions, and after 48 h from aged solutions. For each sampling also a retain sample was taken.
- Sample storage conditions before analysis: All samples were stored deep frozen until they were transferred to the analytical laboratory.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test item is a UVCB consisting of compounds with low water solubilities. Therefore, the test item was applied as water accommodated fraction (WAF) after filtration. A stock solution was prepared by directly weighing 10 mg test item into 1000 mL test medium. This stock solutions was stirred in the dark at room temperature for 24 h (based on the OECD Series on Testing and Assessment N° 23). The solution was clear and transparent. After 24 h of stirring, insoluble particles of the test item were separated from the liquid phase by filtration through a filter with ≤ 0.45 µm mean pore size (according to OECD Series on Testing and Assessment N° 23). Transfer of undissolved test item was avoided. The resulting filtrate was used as application solution. The preparation was repeated after 24 h.
- Controls: 4 replicates without test item.
- Evidence of undissolved material: The solution was clear and transparent.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain/clone: clone V
- Source: Federal Environment Agency, Berlin, Germany
- Age of parental stock (mean and range, SD): Freshly hatched daphnia < 24 h old were used for the test.
- Breeding conditions: D. magna was bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium. The pH-value of the aerated water was within a range of 6.0 - 9.0. The dissolved oxygen was above 60% saturation and the total hardness 140 - 250 mg/L (as CaCO3), corresponding to 7.8 - 14 ° dH. The daphnia were reared at a temperature of 20 ± 2 °C in a climatic chamber with 16 h of illumination and 8 h of darkness. The medium was changed three times per week.
- Food type during breeding: The animals were fed with single cell green algae.
- Frequency: At least three times a week.
- Feeding during test: No.

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES
A pipette was used to separate the young daphnia from the adults.

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Hardness:
13 ° dH = 232 mg/L CaCO3
Test temperature:
20.5 ± 0.9 °C
pH:
7.92 ± 0.09
Dissolved oxygen:
8.8 ± 0.3 mg/L
Nominal and measured concentrations:
control and 10.0 mg/L (nominal)
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL glass beakers filled with 50 mL test medium.
- Renewal rate of test solution: 24 h (1 time during test).
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Culture medium different from test medium: Culture medium same as test medium (Elendt M4).
- Intervals of water quality measurement: The test temperature and the pH-values as well as the oxygen concentration of the test solutions were measured at all test item loading rates at t = 0 h and t = 24 h in fresh test solutions and after t = 24 h and t = 48 h in aged test solutions in one separate replicate per test item loading rate without test organisms.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light/ 8 h dark

EFFECT PARAMETERS MEASURED:
- Immobilisation: after 24 and 48 h

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: Based on the results of a non-GLP range-finding test a limit test with 10.0 mg/L and a control were tested in the definite test.
Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
WAF
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
>= 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
WAF
Basis for effect:
mobility
Details on results:
After 24 and 48 h of exposure no immobilization was observed in the control and 10.0 mg/L loading rate. Since no immobilization occurred during the test, no statistical analysis was performed. The EL50 could not be determined and the NOELR was derived from the highest loading rate tested at which the immobilization was not higher than the allowed control immobilization (≤ 10%).
Results with reference substance (positive control):
- Results with reference substance valid? Yes, the results comply with OECD 202 and are in line with historical data.
- Relevant effect levels: EC50 (24 h) = 1.0 - 2.0 mg/L potassium dichromate
- Other: The reference test dates 20 Sep 2016.
Reported statistics and error estimates:
Since no immobilisation occurred during the test, no statistical analysis was performed.
The NOELR was established based on the highest loading rate at which the immobilisation is not higher than the allowed control immobilisation (≤ 10% immobilisation).

Any other information on results incl. tables

ANALYTICAL RESULTS

The test item is a UVCB consisting of several components with similar chemistry. No adequate method for a sufficiently accurate quantitative analysis of the individual components was available. As indication of the concentration of the test item in the respective test solutions, the content of total organic carbon (TOC) in the solutions was measured. In addition, the TOC content of the test item as a bulk material was determined.

TOC analysis proved the presence of the test item in the solutions but was found to be unsuitable for quantitative purposes. Thus, the effects were evaluated based on the nominal loading rates.

TOC analysis showed that the measured concentrations of the fresh samples ranged from 64.5 – 147% of the nominal loading rate. In the aged samples the measured content was 147% of the nominal loading rate (Table 1).

 

Table 1. TOC measurement.

Test Item Loading rate

Nominal

[mg/L]

TOC content

Nominal

[mg/L]1)

Sampling

TOC measured in test solution

[mg/L]*

% of nominal

Geometric mean [%]

Control

 

0

0 h fresh

24 h fresh

48 h aged

< LOQ

< LOQ

< LOQ

-

-

10.0

 

6.82

0 h fresh

24 h fresh

48 h aged

4.4

10.0

10.0

64.5

147

147

 

125

- = Not calculated; n.d. = not detectable; LOQ = 0.30 mg/L TOC

1)= Based on analysed TOC-content of test item (mass of carbon was 68.2% of total mass)

*= Control corrected.

Applicant's summary and conclusion