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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:
31 October 2017 to 11 November 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

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
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 116B2018
- Physical state: clear and colorless liquid
- Analytical purity: 96.3%
- Expiration date of the lot/batch: 31 August 2018
- Purity test date: 15 Nov 2016
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature container flushed with nitrogen

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: negative control and saturated solution (limit test)
- Volume: 42.5 mL, a complete vial (see sampling method)
- Sampling method: Extra sample vials were made to be sacrificed for analytic monitoring (one for t=0h and one for t=48h).
- Sample storage conditions before analysis: Samples were analyzed on the day of sampling.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test substance is volatile. Therefore, 42.5mL air-tight vials were completely filled with medium and closed with a septum-sealed screw cap. PPVE-2 (2.5 μL) was then injected through the septum with gas-tight syringe. Thereafter the septum was sealed using parafilm. Vials were rotated slowly for seven days, and then centrifuged at 500g for 90 minutes to pull dissolved test substance into a single drop at the bottom of the vial.
- Controls: Blank only
- Evidence of undissolved material: Test solutions contained a visible small droplet of undissolved test substance at the bottom of the vials.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain/clone: Daphnia magna (Crustacea, Cladocera) (Straus, 1820).
- Source: In-house laboratory culture with a known history, at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.
- Age of parental stock (mean and range, SD): > 2 weeks
- Age at study initiation: < 24 hrs
- Feeding during test: No
-Culturing: Adult daphnids were cultured in M7 medium. Adult daphnids in the cultures were held for at least 14 days, and were not more than 4 weeks old prior to collection of the juveniles for testing. During the period preceding the test, water temperatures were 18 - 22°C. Daphnids in the cultures were fed daily a suspension of freshwater algae. The adults were fed prior to test initiation, but neonates were not fed during the test. Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.

Study design

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

Test conditions

Hardness:
180 mg CaCO3 in the medium
Test temperature:
21 - 22 °C
pH:
7.5 - 7.9
Dissolved oxygen:
8.3 - 9.0 mg/L
Nominal and measured concentrations:
Nominal: 100 mg/L
Measured: 1.6 µg/L (average)
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass VOA vials closed with PTFE-faced septa, completely filled. Fill volume ca. 42.5 mL
- Aeration: no
- No. of organisms per vessel:five
- No. of vessels per concentration (replicates): four
- No. of vessels per control (replicates):four
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium prepared using reverse-osmosis purified water (GEON Waterbehandeling, Berkel-Enschot, The Netherlands)
- Intervals of water quality measurement: At the start of the test (0-hour), and at the end (48-hours)
OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light; 8 hours dark
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2CrO7)

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 1.6 µg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
mobility
Remarks on result:
other: 1.6 µg/L was considered the maximum soluble concentration in test medium. Test conducted at a loading rate of 100 mg/L.
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:
- Mobility: no immobilization of test organisms was observed
- Mobility of control: No immobilization observed
- Effect concentrations exceeding solubility of substance in test medium: Yes
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Dose-response test: Yes
- ECx: 48-hr EC50 = 0.54 mg/L (95% CI, 0.49 mg/L to 0.61 mg/L)
- Other: The sensitivity of Daphnia magna originating from the batch used in this test, for potassium dichromate fell within the expected range of sensitivity observed during previous years (n > 60; 48-hr EC50 between 0.3 and 1.0 mg/L.) The reference substance test was conducted 63 days after the testing of PPVE-2.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
the validity criteria of OECD TG 202 were fulfilled. Control immobilization was < 10% at the end of the test. The dissolved oxygen concentration was ≥ 3 mg/L in the control and test vessels at the end of the test.
Conclusions:
There was no immobilization of Daphnia magna in a limit test with PPVE-2 done at a loading of 100 mg/L (OECD TG 202). The measured geometric mean concentration of PPV-2 was 1.6 µg/L.
Executive summary:

The 48-hour EC50 of PPVE-2 to Daphnia magna was examined in a static test conducted according to OECD TG 202. Test vessels were sealed to minimize loss of the volatile substance. Test solutions were prepared as the control (0 mg/L) and at 100 mg/L. Analytically determined concentrations were < LOQ, and 1.6 µg/L, which was considered the maximum soluble concentration in the test medium. Final test substance concentrations were within 60% of initial, and undissolved material was present in the vessels throughout the test. No immobilization of the daphnids was observed by 48 hours. The 48-hour EC50 is > 1.6 µg/L, based on the measured concentration, and > 100 mg/L, based on the loading (nominal) concentration.

The test was conducted according to internationally accepted test guidelines and was GLP compliant. It is reliable without restriction and suitable for Risk Assessment, Classification and Labeling, and PBT Analysis.