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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:
27 June 2018 to 29 June 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
OECD Guidelines for Testing of Chemicals, No.:202 (Adopted: 13 April 2004) OECD: Paris.
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
Commission Regulation (EU) 2017/735 of 14 February 2017 amending, for the purpose of its adaptation to technical progress, the Annex to Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), Annex IV Part C, C.2.
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Version / remarks:
EPA Health Effects Test Guidelines, OCSPP 850.1010.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
See confidential details on test material
Analytical monitoring:
yes
Details on sampling:
Duplicate samples (from each replicate) were taken (2 x ~2 mL) in plastic tubes at the beginning and at the end of the renewal periods from the control and at the applied test concentration level.
Test item was demonstrated to be not stable in ISO media at room temperature. After sampling, samples were frozen immediately and kept approximately at -20°C at the Test Facility. One set of the samples was sent to the Test Site for analysis and analyzed within 27 days since sampling and one set was retained as a back-up at the Test Facility, if required for any confirmatory analyses (discarded after satisfactory results were obtained on the first set of).
Vehicle:
no
Details on test solutions:
The test item is poorly soluble in water (< 1 mg/L), it was not stable in ISO media (based on analytical method validation) and it is suspected to rapidly volatilize from water.
A test solution was prepared using a saturated solution method according to the Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, OECD No. 23.
A saturated test item solution (nominal loading rate of 100 mg/L) was prepared by dispersing/dissolving the amount of test item into the test medium (ISO Medium) a day before the start of the study. This solution was shaken for about 24 hours at approximately 30°C in a tightly closed vessel in order to minimize losses due to evaporation. After 2 hours of standing/equilibration (at approximately 20°C), the non-dissolved test material was removed by centrifugation at 4700 rpm for 10 minutes to give the 100 % saturated solution and then the solution used for the study.
As a Limit test was carried out, further dilution of stock solution was not performed.
The test solution preparation process was conducted in a closed system with reduced headspace in order to avoid potential losses due to volatilization.
Due to the results of the analytical method validation, the test was performed under semi-static conditions. The frequency of the water renewal periods was 24 hours. Prior to treatment of each renewal period, test item solution was prepared by the method described above.
The test solution was prepared just before introduction of the Daphnia (start of the treatments).
Test organisms (species):
Daphnia magna
Details on test organisms:
Species and strain: Daphnia magna
Source: István Szent University, 2100 Gödöllő, Páter Károly u. 1, Hungary
Breeding: The Daphnia are bred in the Ecotoxicological Laboratory of Citoxlab Hungary Ltd. The health of the stock animals is monitored by visual daily checking. Abnormal behaviour or significant decrease of the population is recorded.
Justification of strain: Daphnia magna is the standard species of the acute immobilisation test.
Number of animals: There were 20 animals in test group and control group respectively, divided into 4 replicates (5 animals / replicate)
Age of the animals: They were less than 24 hours old at the beginning of the test
Acclimatization: There was no acclimatization because the water used was similar to the culture water.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Remarks on exposure duration:
As per guideline
Post exposure observation period:
No post exposure observation period.
Hardness:
The reconstituted water (ISO medium) had a total hardness of 249 mg/L (as CaCO3).
Test temperature:
The water temperature was measured at the start and at the end of the renewal periods in each test vessel. The test temperature was in the range of 20.2 – 20.8°C measured in the test vessels.
The additionally measured temperature in the climate chamber was between 20.0 – 20.9°C.
pH:
The pH of the test solution was not adjusted and not varied by more than 1.5 units in any one test. The pH was measured at the start and at the end of the renewal periods in each test vessel and was in the range of 7.58 – 7.73.
Dissolved oxygen:
The dissolved oxygen concentration was measured in each test vessel at the start and at the end of the renewal periods and was in the range of 8.0 – 8.6 mg/L.
Salinity:
Not applicable-Freshwater study
Conductivity:
Not specified
Nominal and measured concentrations:
The biological results are based on both the nominal and measured test concentration.
Details on test conditions:
PERFORMANCE OF THE TEST
The test item is poorly soluble in water (< 1 mg/L), was not stable in ISO media (based on analytical method validation) and it was suspected to rapidly volatilize from water. For this reason the test was conducted in a completely closed system with minimized headspace in the test vessels.

Taking into account that the test item is a highly volatile substance the study was performed in a closed system and followed the OECD recommendations. Therefore, Erlenmeyer flasks with stopper were used instead of glass beakers to minimize the loss of the substance from solution.
The test duration was 48 hours. Twenty animals, divided into four groups (Erlenmeyer flasks with stopper with a volume of 50 mL) of five animals each (~40 mL test solution/flask; 8 mL test solution/animal) were used at the test concentration as well as for the control. The animals were not fed during the test.
Due to the results of the analytical method validation, the test was performed under semi-static conditions. The frequency of the water renewal periods was 24 hours.
The choice of the test concentration was done on the basis of the results of a preliminary range-finding test.

Preliminary Range-Finding Test
A concentration range-finding test was conducted to determine the approximate toxicity of the test item so that appropriate test concentrations could be selected for use in the definitive test. Ten daphnids (divided into two replicates) in each test concentration and control were exposed for 48 hours under semi-static conditions.
During the formulation procedure the stock solution was prepared by the similar method described. The test solutions were prepared by appropriate dilution of the stock solution.

Test item concentrations in the Definitive Test
Because toxic response was not observed during the preliminary concentration range-finding test, a Limit Test was carried out at the solubility level of the test item in the test medium (100 % saturated solution, 100 mg/L nominal loading rate) and one control group in a semi-static system.
The biological results are based on both the nominal and measured test concentration.

OBSERVATIONS
The immobility or mortality of the Daphnia was determined by visual observation 24 and 48 hours after the start of the test. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker are considered to be immobile.
The number of immobilised animals and the percentage of immobility were determined at 24 and 48 hours.
The water temperature, the oxygen concentrations and pH of the control and the test solution were measured at the beginning and at the end of the renewal periods.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
100 other: mg/L loading rate
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
> 100 other: mg/L loading rate
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L loading rate
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
VALIDITY
There were no immobilized animal in the control group and the dissolved oxygen concentration at the end of the test in control and test vessels was more than 3 mg/L.
All validity criteria were within acceptable limits and therefore the study can be considered as valid.
In addition to immobility, no abnormal behaviour or appearance of test animals was detected.
Results with reference substance (positive control):
The 24h EC50: 0.65 mg/L, (95 % confidence limits: 0.61 – 0.70 mg/L)
Reported statistics and error estimates:
No statistical analysis was performed because of the lack of toxic effects.
The EC50, NOEC, LOEC and EC100 values were determined directly from the raw data.

Results of the preliminary Range-Finding Test

Nominal concentrations

[% sat. sol.]

Untreated control

0.1

1

10

100

Number of treated / immobilized animals

10 / 0

10 / 0

10 / 0

10 / 0

10 / 0

 

Number and percentage of immobilised animals

Test Group

Number of treated animals

Number of immobilised animals

24 hours

48 hours

Number

Percent

Number

Percent

Control

100 mg/L

nominal loading rate

20

 

20

0

 

0

0

 

0

0

 

1

0

 

5

 

DATA OF TEST CONDITIONS

Temperature measured in the test vessels (°C)

Test group

Replicate

Measuring

0 h

(fresh media)

24 h

(old media)

24 h

(fresh media)

48 h

(old media)

Control

1

2

3

4

20.8

20.8

20.8

20.8

20.2

20.2

20.2

20.2

20.6

20.6

20.6

20.6

20.3

20.3

20.3

20.3

100 mg/L

nominal loading rate

1

2

3

4

20.8

20.8

20.8

20.8

20.2

20.2

20.2

20.2

20.6

20.6

20.6

20.6

20.3

20.3

20.3

20.3

 

Oxygen concentration measured in the test vessels (mg/L)

Test group

Replicate

Measuring

0 h

(fresh media)

24 h

(old media)

24 h

(fresh media)

48 h

(old media)

Control

1

2

3

4

8.6

8.6

8.6

8.6

8.6

8.6

8.6

8.6

8.4

8.4

8.4

8.4

8.5

8.5

8.5

8.5

100 mg/L

nominal loading rate

1

2

3

4

8.1

8.1

8.1

8.1

8.5

8.5

8.5

8.5

8.0

8.0

8.0

8.0

8.4

8.4

8.4

8.4

 

pH measured in the test vessels

Test group

Replicate

Measuring

0 h

(fresh media)

24 h

(old media)

24 h

(fresh media)

48 h

(old media)

Control

1

2

3

4

7.64

7.64

7.64

7.64

7.58

7.60

7.60

7.60

7.70

7.70

7.70

7.70

7.65

7.65

7.65

7.65

100 mg/L

nominal loading rate

1

2

3

4

7.73

7.73

7.73

7.73

7.70

7.70

7.70

7.70

7.69

7.69

7.69

7.69

7.66

7.66

7.66

7.66

 

DATA OF IMMOBILISATION

Immobilisation of the test animals

Test group

Number of animals

Number of immobilised animals

24 h

48 h

Control

5

5

5

5

0

0

0

0

0

0

0

0

100 mg/L

nominal loading rate

5

5

5

5

0

0

0

0

0

1

0

0

 

Validity criteria fulfilled:
yes
Conclusions:
Acute toxicity of FLUOROSULFONIC ADDUCT was assessed with Acute immobilisation test on Daphnia magna, over an exposure period of 48 hours in a semi-static system.
All validity criteria were met during this study.
Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of FLUOROSULFONIC ADDUCT were the followings:
The 24h and 48h EC50 value: > 100 mg/L nominal loading rate
The 48h EC100 value: > 100 mg/L nominal loading rate
The 48h No-Observed Effect Concentration (NOEC): 100 mg/L nominal loading rate
The 48h Lowest Observed Effect Concentration (LOEC): > 100 mg/L nominal loading rate
Based on the results of this study, the test item FLUOROSULFONIC ADDUCT had no toxic effect at saturation (0.252 mg/L, measured); the EC50 results and the LOEC are higher than the solubility level of the test item in the test medium.
Executive summary:

The acute toxicity of FLUOROSULFONIC ADDUCT on Daphnia magna was assessed in an acute immobilisation test, over an exposure period of 48 hours in a semi-static test system.

 

The test item is poorly soluble in water (< 1 mg/L), was not stable in ISO media (based on analytical method validation) and it was suspected to rapidly volatilize from water. For this reason the test was conducted in a completely closed system with minimized headspace in the test vessels.

 

Because no toxic response was observed during the preliminary range-finding test, a Limit Test was carried out using only one test concentration at the solubility level of the test item in the test medium (100 mg/L nominal loading rate, 100 % saturated solution) and one control group in the definitive test.

The test concentration was analytically determined at the start and at the end of the renewal periods. The corresponding measured geometric mean test concentration was 0.252 mg/L.

 

The biological results are based on both the nominal and geometric mean measured test item concentration.

 

Twenty animals, divided into four groups (Erlenmeyer flasks with stopper) of five animals each were used at the test concentration and for the control.

All validity criteria were met during this study.

 

Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of FLUOROSULFONIC ADDUCT were the followings:

The 24h and 48h EC50 value: > 100 mg/L nominal loading rate

The 48h EC100 value: > 100 mg/L nominal loading rate

The 48h No-Observed Effect Concentration (NOEC): 100 mg/L nominal loading rate

The 48h Lowest Observed Effect Concentration (LOEC): > 100 mg/L nominal loading rate

 

Based on the results of this study, the test item FLUOROSULFONIC ADDUCT had no toxic effect at saturation (0.252 mg/L, measured); the EC50 results and the LOEC are higher than the solubility level of the test item in the test medium.

Description of key information

Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of FLUOROSULFONIC ADDUCT were the followings:

The 24h and 48h EC50 value: > 100 mg/L nominal loading rate

The 48h EC100 value: > 100 mg/L nominal loading rate

The 48h No-Observed Effect Concentration (NOEC): 100 mg/L nominal loading rate

The 48h Lowest Observed Effect Concentration (LOEC): > 100 mg/L nominal loading rate

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
100 mg/L

Additional information

The acute toxicity of FLUOROSULFONIC ADDUCT on Daphnia magna was assessed in an acute immobilisation test, over an exposure period of 48 hours in a semi-static test system.

 

The test item is poorly soluble in water (< 1 mg/L), was not stable in ISO media (based on analytical method validation) and it was suspected to rapidly volatilize from water. For this reason the test was conducted in a completely closed system with minimized headspace in the test vessels.

 

Because no toxic response was observed during the preliminary range-finding test, a Limit Test was carried out using only one test concentration at the solubility level of the test item in the test medium (100 mg/L nominal loading rate, 100 % saturated solution) and one control group in the definitive test.

The test concentration was analytically determined at the start and at the end of the renewal periods. The corresponding measured geometric mean test concentration was 0.252 mg/L.

 

The biological results are based on both the nominal and geometric mean measured test item concentration.

 

Twenty animals, divided into four groups (Erlenmeyer flasks with stopper) of five animals each were used at the test concentration and for the control.

All validity criteria were met during this study.

 

Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of FLUOROSULFONIC ADDUCT were the followings:

The 24h and 48h EC50 value: > 100 mg/L nominal loading rate

The 48h EC100 value: > 100 mg/L nominal loading rate

The 48h No-Observed Effect Concentration (NOEC): 100 mg/L nominal loading rate

The 48h Lowest Observed Effect Concentration (LOEC): > 100 mg/L nominal loading rate

 

Based on the results of this study, the test item FLUOROSULFONIC ADDUCT had no toxic effect at saturation (0.252 mg/L, measured); the EC50 results and the LOEC are higher than the solubility level of the test item in the test medium.

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