2-Pentene, 1,1,1,2,3,4,5,5,5-nonafluoro-4-(trifluoromethyl)-, (E)-

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:

29 Jun - 10 Dec 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study under GLP, analytical confirmation of exposure

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

HFP kinetic dimer

IUPAC Name:

HFP kinetic dimer

Details on test material:

- Name of test material (as cited in study report): MTDID 948, (E)-1,1,1,2,3,4,5,5,5-nonafluoro-4-(trifluoromethyl)pent-2-ene
- Substance type: pure active substance
- Physical state: liquid
- Analytical purity: 99%
- Expiration date of the lot/batch: 09 Aug 2016
- Stability under test conditions: volatile, vapour pressure: 35464Pa at 293K
- Storage condition of test material: at room temperature

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All test concentrations and controls, at t = 0hr and 48 hrs
- Sampling method: Because the substance is extremely volatile, test chambers were not sampled directly. Instead, duplicate, abiotic vials were set up and held under test conditions for each time point. An appropriate volume (0.25 mL to 10 mL) was taken from the test solution and transferred via gas-tight syringe through the septum to a VOA vial closed with a bonded septum cap. The vial had contained sufficient blank medium for a total volume of 10 mL after addition of the sample.
- Sample storage conditions before analysis: None, samples analyzed immediately

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test substance at 100 mg/L in test medium was shaken for seven days at 100 RPM in completely filled, septum-closed VOA vials. Vials were then centrifuged for 90 min at ca. 500xg to consolidate the test substance into a single droplet at the bottom of the vial above the Water Soluble Fraction (WSF). Lower dilutions were made by using a gas-tight syringe to inject precise volumes of the WSF into completely filled, septum-closed VOA vials, with vent needles to allow drainage of blank medium.
- Controls:Blank test medium
- Evidence of undissolved material: Test solutions were clear and colorless except the 100% WSF, which contained a droplet of undissolved test substance.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Straus, 1820
- Source: In-house laboratory culture with known history (at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions).
- Age at study initiation (mean and range, SD): < 24 hours, from parental daphnids of more than two weeks old
- Method of breeding: By acyclical parthenogenesis. Each batch started with newborn daphnids, i.e. less than 3 days old, by placing about 250 organisms into 5 L of M7 medium (based on modified ISO medium, see Test Conditions) in an all-glass culture vessel. Cultures maintained for a maximum age of 4 weeks. After 7 days of cultivation half of the M7 medium was renewed twice per week. Cultures were maintained at 18 - 22 °C and prior to test were fed daily with a suspension of freshwater algae. Stock, 2nd to 5th brood, showed 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.
- Feeding during test: No

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

vials were completely closed during the test.

Test conditions

Hardness:

180 mg CaCO3/L

Test temperature:

19.4 - 21.0 °C

pH:

7.8 - 8.0

Dissolved oxygen:

8.5 - 9.3 mg/L

Nominal and measured concentrations:

Nominal: Control (0 mg/L), 4.6%, 10%, 22%, 46% and 100% of the saturated solution (i.e., 100 mg/L)
Measured:

Details on test conditions:

TEST SYSTEM
- Test vessel: 42.5 mL glass VOA vial with PTFE-faced septum, completely filled
- Aeration: No
- 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: Water used to prepare media was tap water purified by reverse-osmosis (RO-water, GEON Waterbehandeling, Berkel-Enschot, The Netherlands)
- Culture medium different from test medium: Yes.
Test medium: Adjusted ISO medium.
CaCl2∙2H2O 211.5 mg/l
MgSO4∙7H2O 88.8 mg/l
NaHCO3 46.7 mg/l
KCl 4.2 mg/l
Culture medium: M7; M7 as used in this laboratory is prepared using a slightly modified ISO medium supplemented with macronutrients, micronutrients and vitamins as per standard M7 medium (OECD 202).

OTHER TEST CONDITIONS
- Adjustment of pH: None
- Photoperiod: 16h light, 8h dark
- pH and dissolved oxygen were measured at the beginning and ending of the test, temperature was measured continuously in a temperature control vessel

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : At 24h and 48h, immobility (including mortality) was measured.

TEST CONCENTRATIONS
- Range finding study: yes
- Test concentrations: Control, 0.1%, 1.0% and 10% dilutions of WSF plus 100% WSF with undissolved test substance
- Results used to determine the conditions for the definitive study: yes, 40% effect with 10% dilution (average initial concentration, 16 µg/L), 100% effect with undiluted WSF (not analyzed)

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

14 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Remarks:

trans isomer

Basis for effect:

mobility

Remarks on result:

other: 95% CI, 10-19 µg/L

Details on results:

- Other biological observations: In rangefinder, no organisms were trapped at undissolved test substance deposit.
- Mortality of control: 1 of 20
- Other adverse effects control: No
- Abnormal responses: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50: 0.38 mg/L. Historical range for the reference substance at the contract lab lies between 0.3 and 1.0 mg/L
- Other: Reference substance toxicity assay conducted 5 days after test substance.

Reported statistics and error estimates:

The 24 and 48h-EC50-value was calculated from the probits of the percentages of affected daphnids and the logarithms of the corresponding test substance concentrations using the maximum likelihood estimation method.
ToxRat Professional v 3.0.0 (ToxRat Solutions® GmbH, Germany) was used to perform the analyses.

Any other information on results incl. tables

Table 2, Immobilization during the Daphnia test (20 per concentration)
Average exposure concentration	Total immobilized at τ = 24 hours	Percent Effect at τ = 24 hours	Total immobilized at τ = 48 hours	Percent effect at τ = 48 hours
Control	0	0	1	5
1.4	0	0	0	0
2.8	0	0	1	5
5.9	0	0	1	5
17	6	30	12	60
63	14	70	20	100

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

<10% immobilization in controls, DO >3mg/L in all test vessels

Conclusions:

The EC50 of HFP kinetic dimer to D. magna is 14 µg/L (95% CI, 10 - 19 µg/L).

Executive summary:

Toxicity of HFP kinetic dimer to D. magna was assessed in a test conducted according to OECD 202. The test substance was not soluble at the limit of 100 mg/L and is extremely volatile. Water soluble fractions at 100 mg/L were made in closed VOA vials with PTFE-face septum closures. Dilution series were made using gas-tight syringes into further VOA vials. Four replicates were prepared of negative controls and at each concentration, with analytical determination of initial and final test substance concentration. Vials at the highest loading rate intentionally contained undissolved test material in a droplet which was formed by centrifugation before addition of daphnids. Test substance concentration increased at the highest loading rate during the test, but decreased at all other concentrations. The EC50 based on geometric mean concentration was 14 µg/L (95% CI, 10 - 19 µg/L).

The test was conducted according to an internationally accepted guideline and was GLP compliant. Test substance concentrations were analytically confirmed. This test is considered reliable without restrictions. It is suitable for Risk Assessment, Classification & Labeling, and PBT Analysis.