Acetic acid, 2,2-difluoro-2-[[2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy]-, ammonium salt (1:1)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-04-20 until 2009-06-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline-conform study under GLP without deviations.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss Federal Office of Public Health, 2008-11-12

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/l of dry salt of test item cC604 (i.e. 625 mg/l of test item cC604, which contains 16% active ingredient)
- Sampling method: Duplicate samples were taken from each treatment before the start of the test and at the end of the test after 48 hours.
- Sample storage conditions before analysis: All samples were stored deep-frozen (at about -20 °C) immediately after sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The test medium was prepared by mixing 3124.8 mg of the test item into 5 liters of test water using ultrasonic treatment for 15 minutes and intense stirring for 15 minutes at room temperature. The test medium was prepared just before introduction of the fish (i.e., start of the test).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Strain: Brachydanio rerio
- Source: The fish were obtained from a breeding culture at Harlan Laboratories.
- Age at study initiation (mean and range, SD): no data
- Length at study initiation (length definition, mean, range and SD): 3.1 ± 0.12 cm (Mean ± SD)
- Weight at study initiation (mean and range, SD): the mean body wet weight was 0.25 ± 0.06 g (Mean ± SD).
- Method of breeding: During holding and acclimatization until one day before the start of the test, the fish were fed with a commercial fish diet (Tetra Min). No further details given.
- Feeding during test
- Food type: no feeding during the 96 h testing period
- Amount: no feeding during the 96 h testing period
- Frequency: no feeding during the 96 h testing period


ACCLIMATION
- Acclimation period: one week
- Acclimation conditions (same as test or not): Prior to test start, the test fish were acclimated for one week to the test water and temperature.
- Type and amount of food: During holding and acclimatization until one day before the start of the test, the fish were fed with a commercial fish diet
- Feeding frequency: no data
- Health during acclimation (any mortality observed): During the last two weeks prior to the test, no fish died in the test fish batch and all fish were healthy.


QUARANTINE (wild caught)
not applicable

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

no data

Hardness:

1.25 mmol/l as CaCO3

Test temperature:

20-21 °C

pH:

7.5 - 7.6

Dissolved oxygen:

7.9 mg/L or higher

Salinity:

No data, however the detailed mineral composition of the reconstituted water is given in the study report.

Nominal and measured concentrations:

The nominal concentration of 625 mg test item cC6O4/L, corresponding to 100 mg dry salt cC604/L (test item contains 84% water).
Concnetration of cC6O4 (calculated based on the measurement of the content of dry salt) in the test medium varied in the range from 81 to 103% of the nominal value.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: fill volume 5 l, no further data
- Aeration: The test vessels were slightly aerated during the test period.
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): not applicable
- No. of organisms per vessel: 7 fish were introduced into each test vessel in a random order
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): not applicable
- Biomass loading rate: 0.35 g fish wet weight per liter test medium


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water was used in the study. It consisted of analytical grade salts dissolved in purified water to obtain nominal concentrations accc. to the OECD guideline.
- Total organic carbon: not applicable
- Particulate matter: not applicable
- Metals: Na, K, Ca, Mg only
- Pesticides: not applicable
- Chlorine: not applicable
- Alkalinity: 0.4 mmol/l
- Ca/Mg ratio: 4:1 based on molarity
- Conductivity: No data, however the detailed mineral composition of the reconstituted water is given in the study report.
- Culture medium different from test medium: yes, acclimatization of test organisms included in the study
- Intervals of water quality measurement: The concentration of dry salt of the test item cC6O4 was analytically measured in the duplicate test medium samples of the single test concentration taken at the start, after 48 hours and end of the test. From the control, only one of the duplicate samples was analyzed from each of the sampling times. The water temperature, pH and dissolved oxygen concentrations were measured at the start of the test and once daily during the test for each treatment.


OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: A 16 hour light to 8-hour dark photoperiod, with a 30-minute transition period was used
- Light intensity: 100 to 600 Lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The test fish were observed for mortality and visible abnormalities.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable
- Justification for using less concentrations than requested by guideline: This limit test was based on a range-finding test and was performed in compliance with the test guidelines to demonstrate that the test item has no toxic effect on fish up to at least this concentration.
- Range finding study
- Test concentrations: no data
- Results used to determine the conditions for the definitive study: The limit test was based on the results of a range-finding test (non-GLP).

Reference substance (positive control):

not required

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: none reported
- Observations on body length and weight: none reported
- Other biological observations: none reported
- Mortality of control: no mortalities reproted
- Other adverse effects control: none reported
- Abnormal responses: none reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none reported
- Effect concentrations exceeding solubility of substance in test medium: none reported

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

not statistics applied

Sublethal observations / clinical signs:

Tab. 1: Mortality and Visible Abnormalities Observed in the Test Fish

Dry salt conc. (mg/l)	Number of abnormal and dead fish
	Observation time (h)
	3	24	48	72	96
Control	0 / 0	0 / 0	0 / 0	0 / 0	0 / 0
100	0 / 0	0 / 0	0 / 0	0 / 0	0 / 0
LC-50	>100	>100	>100	>100	>100

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, the test item had no acute toxic effects on zebra fish up to the nominal concentration of 100 mg dry salt cC6O4 ammonium salt/L (corresponding to 625 mg test item cC6O4 ammonium salt/L) under the present test conditions.

Executive summary:

The acute toxicity of the test item cC6O4 ammonium salt to zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to the EU Commission Directive 92/69/, Part C.1 and Commission Regulation (EC) No 440/2008, Part C.1, as well as according to the OECD Guideline for Testing of Chemicals No. 203 (1992).

A limit test was performed in accordance with the guidelines to demonstrate that the test item has no toxic effect on the test organisms at a nominal concentration of 625 mgcC6O4 ammonium salt/L corresponding to 100 mg dry salt cC6O4 ammonium salt/L (test item contains 84% water). Thus, a single nominal concentration of the test item was tested. Additionally, a control group was tested in parallel. In the control and at the test concentration of 100 mg dry salt/L, no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours.

In conclusion, the test item had no acute toxic effects on zebra fish up to the nominal concentration of 100 mg dry salt cC6O4 ammonium salt/L (corresponding to 625 mg test item cC6O4/L) under the present test conditions.

Description of key information

The acute toxicity of the test item cC6O4 ammonium salt to zebra fish (Brachydanio rerio) was determined in a 96-hour static test according to the EU Commission Directive 92/69/, Part C.1 and Commission Regulation (EC) No 440/2008, Part C.1, as well as according to the OECD Guideline for Testing of Chemicals No. 203 (1992). The study was performed under GLP without deviations.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The test item had no acute toxic effects on zebra fish up to the nominal concentration of 625 mg test item cC6O4 ammonium salt/L corresponding to 100 mg dry salt cC6O4 ammonium salt/L (test item = aqueous solution of cC6O4 ammonium salt containing 84% water).

LC50-96 hours > 100 mg/l