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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 Nov - 27 Nov 2018
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
adopted 13 Apr. 2004
Deviations:
yes
Remarks:
See section "Any other information on material and methods incl tables"
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
adopted 30. May 2008
Deviations:
yes
Remarks:
See section "Any other information on material and methods incl tables"
GLP compliance:
yes (incl. certificate)
Remarks:
Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht Rheinland-Pfalz, Mainz, Germany
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Control and test item concentration
- Sampling method: Concentration was measured at the beginning and at the end of the test and on at medium renewal after 24 h in the new and the old solutions.
- Sample storage conditions before analysis: Samples were frozen
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Differential loading: At test start a saturated solution was prepared by mixing the nominal load of 100.0 mg/L with the corresponding amount of dilution water and shaking vigorously for 24 h and 15 min. The resulting solution was filtrated through 0.45 μm cellulose filters.
- Controls: Yes, medium only
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Source: Umweltbundesamt Berlin (In-house breeding since 27 Sep 2007)
- Feeding during test: No

ACCLIMATION
- Acclimation conditions: Medium was different: M4-medium (husbandry) compared to dilution water (test)
- Type and amount of food: Green algae (Desmodesmus subspicatus) for stock culture
- Feeding frequency: Twice a week

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: 21 h and 5 min before the start of the test, the adult animals were separated from the young. 0.5 h before test start, the adults were caught with the help of a glass tube and the newborn daphnia, aged at test start between 30 min and 21 h and 5 min, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
2.502 mmol/L corresponding to 250 mg CaCO3/L
Test temperature:
20.9 – 22.4 °C
pH:
7.5 - 7.8 (Control)
7.5 - 7.8 (Test item concentration)
Dissolved oxygen:
8.8 - 9.5 mg/L (Control)
8.7 - 9.3 mg/L (Test item concentration)
Nominal and measured concentrations:
Nominal: control and 100 mg/L
Measured: see tables 2 in section "Any other information on results incl tables"
Details on test conditions:
TEST SYSTEM
Test vessel:
- Type: open
- Material, size, headspace, fill volume: glass, 50 mL, headspace and fill volume not given
- Renewal rate of test solution (frequency/flow rate): Renewal after 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water (Daphnia medium) with the following specification was used: CaCl2*2H2O: 293.8 mg/L; MgSO4*7H2O: 123.3 mg/L, NaHCO3: 64.8 mg/L; KCl: 5.8 mg/L;
- pH 7.7
- Culture medium different from test medium: Yes, Dilution water (Test), M4-Medium (recipe of ELENDT for husbandry), Switching from M4-medium (husbandry) to dilution water (test) has been shown not to cause any detrimental effects for test daphnia.
- Intervals of water quality measurement: At test start and test end and medium renewal

OTHER TEST CONDITIONS
- Photoperiod:16/8 h
- Light intensity: Neon tubes

EFFECT PARAMETERS MEASURED: Mobility was measured every 24 h. Daphnia are considered immobilised if they perform no movements or are only able to move their antennae when the beaker is gently agitated.

RANGE-FINDING STUDY: The concentrations to be tested were based on the result of a non-GLP pre-test.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 11.28 µg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Mortality of control: No mortality in the control
- Other adverse effects control: No effect was observed
Results with reference substance (positive control):
The 24 h EC50 value was determined as 1.9 mg/L. The value was within the requested range for the 24 h EC50 between 0.6 - 2.1 mg/L of potassium dichromate K2Cr2O7 as specified in the OECD guideline.
Reported statistics and error estimates:
None of the animals was immobilised in the treatments and the blank control. Therefore, no statistical evaluation was performed. The results were stated based on the geometric mean of the measured concentrations and additionally based on the nominal concentrations.

Biological Results:

Table 1: Immobility

Nominal Concentration

Immobility 24 hours

Immobility 48 hours

 

[mg/L]

absolute

 in %

absolute

 in %

Blank control

0

0

0

0

0

0

0

0

0

0

100

0

0

0

0

0

0

0

0

0

0

 

Analytical Results

Table 2: Measured Concentrations

Nominal Concentration Test Item

Measured concentration test item

 μg/L

mg/L

t = 0 h

t = 24 h old

t = 24 h new

t = 48 h

Blank control

< LOD

--

--

< LOD

100

17.08

5.03

18.90

9.96

100 (additional)

65.30

--

--

--

LOD / LOQ (Limit of detection / quantification) = 1.00 μg/L (lowest calibration level)

 

Table 3: Validity criteria

Criterion from the guideline

Outcome

Validity criterion fulfilled

In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.

No mortality

Yes

The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.

8.8 - 9.5 mg/L

Yes

Validity criteria fulfilled:
yes
Remarks:
For further details please refer to “Any other information on results incl. tables”.

Description of key information

EC50 (48h) >11.28 µg/L (Daphnia magna, OECD 202, measured concentration)

Key value for chemical safety assessment

Additional information

One experimental study is available investigating the short-term toxicity of 4-(octadecylamino)-4-oxoisocrotonic acid (CAS 3077-27-8) to aquatic invertebrates which was performed according to OECD 202 and GLP (LAUS GmbH 2018a). Daphnia magna was exposed to the test item in a limit test to a nominal concentration of 100 mg/L for 48 h under semi-static exposure conditions. The test solution was prepared by adding appropriate amounts of test item to test medium, followed by intensive stirring and filtration. Exposure concentrations were verified by HPLC/MS analysis and the measured concentration was 17.08 μg/L and 18.90 μg/L in the freshly prepared solutions and 5.03 μg/L and 9.96 μg/L in the aged solutions, respectively. Due to the low recovery rate the results are given based on the geometric mean of the measured concentrations. Since no effects were observed the EC50 (48 h) was determined to be >100 mg/L (nominal) and >11.28 µg/L (geometric mean measured concentration).

The discrepancy between the low recovery in the test solutions and the water solubility may have been caused by the different test media, pH-values and temperature.