Dodecanoic acid, 3-[[2-(2-hydroxyethoxy)ethyl]imino]-2,2-dimethylpropyl ester

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

For justification for type of information please refer to the Read-Across Justification attached to IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Duration:

24 h

Dose descriptor:

NOELR

Effect conc.:

0.32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

0.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

1.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: exact value after linear extrapolation: 1.02 mg/L

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

For justification for type of information please refer to the Read-Across Justification attached to IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2009

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:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Version / remarks:

1996

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

A supersaturated test item solution (S.S. = Stock solution) was prepared by dispersing/dissolving the test item amount (nominal load of 100 mg/L) without the use of any organic solvent into the Test Medium (ISO Medium) two days before the start of the renewal periods (on day -2 and day -1). This solution was shaken for 24 hours at app. 30 °C, then was left settling for 24 hours at app. 20 °C and thereafter filtrated through a 0.45 μm filter. Respective filtrates were used for the test.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain: Daphnia magna (Straus)
- Source: National Institute of Public Health, Hungary
- Age at study initiation: Less than 24 hours old
- Feeding during test
- Food type: centrifuged algae suspension (Pseudokirchneriella subcapitata)
- Amount: a few mL
- Frequency: daily

ACCLIMATION
Brood Daphnids were maintained in ISO medium under test conditions prior to the start of the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation period.

Hardness:

(CaCO3) = 148 mg/L

Test temperature:

18 - 22 °C

pH:

pH 7.55 to 7.72

Dissolved oxygen:

6.7 to 6.9 mg/L

Nominal and measured concentrations:

The following nominal concentrations were used in the main study: S.S./1; S.S./2; S.S./4; S.S./8 and S.S./16 (S.S.: supersaturated Stock Solution >> limit of solubility of the test item in the media used). The corresponding calculated test item concentrations were 6.25; 12.5; 25.0; 50.0 and 100.0 mg/mL

Details on test conditions:

- Test units: Glass beakers of 40 mL test medium were used
- Test environment: Climate chamber
- Light period: 8 h dark
- Test water /media: ISO medium (during the holding period and during the test, according to OECD 202).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

125.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % conf. limits: 99.6 – 399.5 mg/L

Key result

Duration:

48 h

Dose descriptor:

NOEC

Remarks:

(EC0)

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The 48h-NOEC (EC0) after the exposure period of 48 hours of Sika Hardener LG to Daphnia magna, was determined to be 50.0 mg/L and the 48h-LOEC was determined to be 100.0 mg/L. The 48h-EC50 was calculated to be 125.5 mg/L and the 48h-EC100 was determined to be >100.0 mg/L.

Results with reference substance (positive control):

Results with reference substance assured that the study is valid.

Reported statistics and error estimates:

The EC50 values of the test item and their confidence limits were calculated using Probit analysis. The analysis was done using the statistical software program “SPSS PC+”. The NOEC (EC0), the LOEC and the EC100 were determined directly from the raw data.

Validity criteria fulfilled:

yes

Conclusions:

In this 48-hour acute toxicity test to Daphnia magna with Sika Hardener LG the 48-h EC50 value was 125.5 mg/L and the 48-h NOEC was determined to be 50.0 mg/L (based on nominal concentrations).

Executive summary:

The purpose of this study was to evaluate the influence of the test item Sika Hardener LG on the mobility respectively survival of Daphnia magna toxicity to aquatic invertebrates study according to EU-method C.2 and OECD guideline 202. Young Daphnia were exposed in a semi-static test to the test item for 48 hours, added to test water at concentrations of 6.25; 12.5; 25.0; 50.0 and 100.0 mg/L.

The 48h-NOEC (EC0) was determined to be 50.0 mg/L, the 48h-EC50 was calculated to be 125.5 mg/mL and the 48h-EC100 was determined to be >100.0 mg/mL.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2004-08-18 to 2005-08-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study, OECD guideline Read-across to hydrolysis product.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

adopted 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sampling preparation:
Aqueous samples (10.0 mL) were membrane filtrated (Nylon, 0.45 μm). The filtrate was acidified with 2M-HCl to pH 2 – 3. 10.0 mL of the filtrate were given on the top of a column with 5.0 g Extrelute. After 30 minutes, elution was slowly performed with ethyl acetate (50 mL, 30 minutes), the eluate was rotated down to dryness and 500 μL ISTD-solution were added. At last, 400 μL of the solution were mixed with 50 resp. 100 μL SIL -Mix in vials, and silylation was performed at 80 °C for 60 minutes.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
1. Experiment:
- Method:
As solubility lies below 100 mg/L, the water-accommodated fraction was prepared for the test. This was done by weighing the nominal load of 100 mg/L, adding the corresponding amount of dilution water and shaking vigorously for 24 hours. The resulting solution was filtrated through 0.45 μm filters.
- Differential loading: 100 mg/L
- Controls: four vessels, each containing 20 mL dilution water and 5 daphnia
2. Experiment:
- Method:
The water-accommodated fraction was prepared for the test. This was done by weighing the nominal load of 100 mg/L, adding the corresponding amount of dilution water and stirring slowly for 24 hours. The solution was left to stand for half an hour. Then the lower phase was used for the test.
- Differential loading: 100 mg/L
- Controls: two vessels, each containing 20 mL dilution water and 5 daphnia
3. Experiment:
- Method:
A stock solution containing 1 g/L in acetone was prepared. This solution was used to prepare the treatment. 100 μL of the acetonic stock solution per litre were used.
- Differential loading: 0.1 mg/L
- Controls: two vessels, each containing 20 mL dilution water with 100 μL/L acetone and 5 daphnia
4. Experiment:
- Method:
A stock solution containing 10 g/L in acetone was prepared. This solution was used to prepare the treatment. 100 μL of the acetonic stock solution per litre were used.
- Differential loading: 1 mg/L
- Controls: two vessels, each containing 20 mL dilution water with 100 μL/L acetone and 5 daphnia
5. Experiment:
- Method:
A stock solution containing 10 g/L in acetone was prepared. This solution was used to prepare the treatment. 100 μL of the acetonic stock solution per litre were used.
- Differential loading: 1 mg/L
- Controls: four vessels, each containing 20 mL dilution water with 100 μL/L acetone and 5 daphnia
6. Experiment:
- Method:
A stock solution containing 10 g/L in acetone was prepared. This solution was used to prepare the treatment. 100 μL of the acetonic stock solution per litre were used at the most.
- Differential loading: 0.1 / 0.18 / 0.32 / 0.56 / 1 mg/L
- Controls: four vessels, each containing 20 mL dilution water with 100 μL/L acetone and 5 daphnia

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Berlin
- Source: Umweltbundesamt Berlin
- Age at study initiation: between 0 and 24 hours
- Feeding during test: none

Test type:

other: 1 + 2 Experiment: static; 3 - 6 Experiment: semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

1. 21.5 – 21.7 °C
2. 20.5 – 20.8 °C
3. 20.2 – 20.8 °C
4. 20.2 - 20.8 °C
5. 20.2 – 20.8 °C
6. 20.2 – 20.8 °C

pH:

1. 7.8 to 8.1
2. 7.8 to 8.0
3. 7.8 to 8.1
4. 7.8 to 8.0
5. 7.8 to 8.1
6. 7.8 to 8.1

Dissolved oxygen:

above 8 mg/L

Nominal and measured concentrations:

1. Experiment: 100 mg/L (nominal)
2. Experiment: 100 mg/L (nominal)
3. Experiment: 0.1 mg/L (nominal)
4. Experiment: 1 mg/L (nominal)
5. Experiment: 1 mg/L (nominal)
6. Experiment: 0.1, 0.18, 0.32, 0.56 and 1 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers, nominal volume 50 L, tall shape
- No. of organisms per vessel: 5 daphnia
- No. of vessels per concentration (replicates): 2 - 4
- No. of vessels per control (replicates): 2 - 4

TEST MEDIUM / WATER PARAMETERS
in accordance to the guideline

OTHER TEST CONDITIONS
- Photoperiod: 16/8 hours, using neon tubes
- Light intensity: 20 ± 2 °C

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

0.32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

1.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: exact value after linear extrapolation: 1.02 mg/L

Details on results:

1 Experiment:
The daphnia were immobilized caused by an oily film on the surface. During the shaking period for the preparation of the WAF, an emulsion was produced. This emulsion couldn’t be separated by membrane filtration and during the test, the undissolved part of the test item rose to the surface. The effect on the daphnia can be stated as a physical effect. Therefore the experiment was repeated, but the test solution was prepared by stirring instead of shaking.
2 Experiment:
The daphnia were immobilized caused by an oily film on the surface. Obviously the undissolved parts of the test item couldn’t be separated by taking the lower phase. The effect on the daphnia can be stated as a physical effect. Therefore the experiment was repeated, but the test solution was prepared by spiking dilution water with a stock solution of the test item in acetone.
3. Experiment:
None of the daphnia were immobilized. As the solubility lies between 0.1 and 1 mg/L (due to the hydrolysis of the test item, no exact water solubility could be determined), it could not be confirmed that the limit of solubility was tested in this experiment. Therefore a further experiment with the treatment 1 mg/L was performed.
4. Experiment:
In the treatment after 24 hours, one daphnia was immobilized at the surface. Through the medium renewal, the daphnia was able to remobilize during the test. Based on these results, a further experiment with analytical determination was performed. In order to ensure testing at the maximal water solubility, the treatment 1 mg/L was chosen.
5. Experiment:
In the treatment 10 % of the daphnia were immobilized on the surface after 24 hours and 30 % after 48 hours. Therefore a further experiment with five concentrations was performed.
6. Experiment:
The immobility values changed in the same treatment between 24 and 48 hours. Caused by the medium renewal some of the daphnia were able to remobilize. In the concentrations 1.0 and 0.56 mg/L an thin oily film could be observed. This film has probably caused the immobilization. As a dose-response correlation is visible, this test was used to determine the results.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50: 1.9 mg/L

Reported statistics and error estimates:

The estimation of the EC50 of the test item was accomplished using the software OriginTM. The data were evaluated using linear fit on a probability-logarithmic scale.

In the fifth experiment at the beginning, after 4, 24 and 28 hours of the test, the content of the test item in each test solution was determined using GC. The measured concentration of each fresh test solution (medium renewal after 24 hours) was 45 and 54 % of the nominal concentration. This is caused by the hydrolysis of the test item which is faster than the average time needed for sample preparation. Four hours after the preparation of each new medium only 10 and 15 % of the nominal concentration were determined. The test item is not stable in water (as has been demonstrated in the respective study “water solubility determination”). Due to the fast degradation referring to determined values is not meaningful and only the nominal values were used for the determination of the biological results.

In the sixth experiment at the beginning, after 4, 24 and 28 hours of the test, the content of the test item in each test solution was determined using GC.

The measured concentrations of the fresh test solutions (at the start of the test and at the medium renewal after 24 hours) were between 21 and 66 % of the nominal concentrations. Sample preparation and clean-up requires about one hour. During this time, the test item undergoes already hydrolysis which probably caused these low recoveries. The measured concentrations 4 hours after preparing each new medium (4 and 28 hours after the start of the test) were between 8 and 35 % of the corresponding nominal concentration. The test item is not stable in water (as has been demonstrated in the respective study “water solubility determination”). Due to the fast degradation referring to determined values is not meaningful and only the nominal values were used for the determination of the biological results.

Validity criteria fulfilled:

yes

Conclusions:

The 48-h EC50 was determined about 1 mg/L (exact value after liner extrapolation: 1.02 mg/L) and the 48-h NOEC was 0.18 mg/L.

Executive summary:

The study was performed in order to evaluate the toxic potential of 2,2-Dimethyl-3-lauroyloxy-propanal towards freshwater shrimp, using the species Daphnia magna. Six experiments were performed. As the test item is poorly water soluble, the “water-accommodated fraction” was tested in the first experiment. This was done by weighing the nominal load 100 mg/L, adding the corresponding amount of dilution water and shaking vigorously for 24 hours. The resulting solution was filtrated through 0.45 μm filters. The daphnia were immobilized on the surface already after 24 hours caused by an oily phase of the test item. The second experiment was performed likewise but with stirring instead of shaking. The resulting solution was left to stand for about 30 minutes. The lower phase was used for the test which was performed as a semi-static test. After 24 hours, the daphnia were immobilized on the surface. The oily film was visible again. All following experiments were performed under semi-static conditions with medium renewal after 24 hours. For the third and fourth experiment, a stock solution in acetone was prepared. The treatments 0.1 and 1 mg/L were prepared by spiking the corresponding amount of dilution water with this stock solution. The treatment 1 mg/L was used as lying well above maximal water solubility of 0.227 mg/L. No daphnia were immobilized after 48 hours. In the control, the same concentration of acetone didn’t show any effect. Based on these experiments, the fifth experiment was performed as a limit-test using the treatment 1 mg/L with an acetone-spiked test solution. After 48 hours 30 % of the daphnia were immobilized on the surface. Therefore the last experiment was performed using five concentrations between 0.1 and 1 mg/L. Twenty daphnia were exposed to the test item for 48 hours in a semi-static test system. After 24 and 48 hours, the immobilized daphnia were counted. As no daphnia were dead after 24 hours, but immobilized on the surface, some of them were able to remobilize during the test. This was potentially due to the test item’s physicochemical properties causing film-formation at the top of the water phase (as noted at higher concentrations). None of the animals were immobilized in the control. The 48 h EC50 was determined about 1 mg/L (exact value after liner extrapolation: 1.02 mg/L) and the 48 h NOEC was 0.18 mg/L. These data were already submitted in a NONS dossier under Directive 92/32/EEC (notification number 05 -04 -1922 -00 from 2005 -10 -25) and considered valid and uncritical from the German Competent Authority (BAUA).

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2009-06-22 to 2009-06-24

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Read-across from analoguos substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Version / remarks:

1996

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

The test item solution was prepared according to the WAF method (Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, section 3.1.2 Media preparation methods, Direct addition. OECD Series Testing and Assessment No.23, Paris September 2000).
A supersaturated test item solution was prepared by dispersing/dissolving the respective test item amount without the use of any organic solvent into the Test Medium (ISO Medium) two days before the start of the renewal periods (on day -2 and day -1). This solution was shaken for 24 hours at app. 30 °C, then was left settling for 24 hours at app. 20 °C and thereafter filtrated through a 0.45 μm filter.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain: Daphnia magna (Straus)
- Source: National Institute of Public Health, Hungary
- Age at study initiation: Less than 24 hours old
- Feeding during test
- Food type: centrifuged algae suspension (Pseudokirchneriella subcapitata)
- Amount: a few mL
- Frequency: daily

ACCLIMATION
Brood Daphnids were maintained in ISO medium under test conditions prior to the start of the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

(CaCO3) = 148 mg/L

Test temperature:

20.2 – 20.7 °C

pH:

pH 8.02 to 8.15

Dissolved oxygen:

6.7 to 6.9 mg/L

Nominal and measured concentrations:

The following nominal concentration was used in the main study: 100.0 mg/L.

Details on test conditions:

Test Units
Type and Size: Glass beakers of 40 mL test medium
Identification: Each test unit was uniquely identified with study code, treatment and replicate number.

Test Conditions
Test Environment: Climate chamber
Temperature: 18 – 22 °C with a maximum deviation of ± 1 °C
Light period: 16 h light : 8 h dark
Recording: Test conditions were measured with suitable instruments and documented in the raw data.

Test water
Reconstituted Water: ISO medium (during the holding period and during the test).
The hardness (CaCO3) of water was: 148 mg/L.
The ISO medium (according to OECD 202) was prepared by adding 25 mL each of the following stock solutions to deionised water.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The 48h-NOEC (EC0) after the exposure period of 48 hours of Sika Hardener LI to Daphnia magna, was determined to be 100.0 mg/L and the 48h-LOEC was determined to be >100.0 mg/L. The 48h-EC50 was calculated to be >100.0 mg/L and the 48h-EC100 was determined to be >100.0 mg/L.

Results with reference substance (positive control):

For the evaluation of the quality of the Daphnia clone and the experimental conditions, potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.
The date of the last study (Study Code: 09/045-023DA) with reference item was: 10 – 11 March 2009. The results of the reference study are the followings:
The 24h-EC50 value: 0.84 mg/L
95 % confidence limits: 0.72 – 0.98 mg/L
In the test with the reference item the results met the expected toxicity of the Potassium dichromate (the 24h EC50 value of the potassium dichromate should be within the range 0.6 mg/L to 2.1 mg/L). The results are based on the nominal concentrations.

Reported statistics and error estimates:

The EC50 values and their 95 %-confidence limits were not calculated by statistical analysis due to the lack of immobilisation. The NOEC (EC0), the LOEC and the EC100 were determined directly from the raw data.

Influence of test item on the mobility of Daphnia magna

Nominal concentration	No. of Daphnia tested	No. of immobilised Daphnia after		% of immobilised Daphnia after
		24 h	48 h	24 h	48 h
Control	20	0	0	0	0
100 mg/L	20	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

A water accommodated fraction (WAF) of 100 mg/L was prepared with Sika Hardener LI. In this 48-hour semi-static acute toxicity test with Daphnia magna the 48 h EC50 value was > 100 mg/L, and the 48 h NOEC was determined to be 100 mg/L.

Executive summary:

The purpose of this study was to evaluate the influence of the test item Sika Hardener LI on the mobility respectively survival of Daphnia magna. Young Daphnia were exposed in a semi-static test to the test item for 48 hours.

A supersaturated test item solution was prepared by dispersing the test item amount (nominal load of 100 mg/L) into the test medium two days before the start of the renewal periods (on day -2 and -1). This solution was shaken for 24 hours at app. 30 °C, then was left settling for 24 hours at app. 20 °C and thereafter filtrated through a 0.45 μm filter. The nominal concentration tested was 100.0 mg/L.

In this 48-hour semi-static acute toxicity test with Daphnia magna the 48 h EC50 value was > 100 mg/L, and the 48 h NOEC was determined to be 100 mg/L. The results were based on the nominal concentrations.

Description of key information

Sika Hardener LG was assessed in a short-term toxicity to aquatic invertebrates study according to EU-method C.2 and OECD guideline 202. Young Daphnia (Daphnia magna) were exposed in a semi-static test to water accommodated fractions (WAF) at nominal concentrations of 100 mg/L. The 48-h EC50 was determined to be 125.5 mg/L and the 48-h NOEC at 50 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

125.5 mg/L

Additional information

Due to the high reactivity of the substance in water, a weight of evidence approach using data form the parent molecule, its two primary hydrolysis products and from an analogous substance were assessed in line with Annex XI REACH Regulation and oECD Guidance No. 23 for evaluation of substances instable in water:

In the study with the parent compound Sika Hardener LG, Young Daphnia were exposed in a semi-static test to the test item for 48 hours, added to test water at concentrations of 6.25; 12.5; 25.0; 50.0 and 100.0 mg/L. The 48h-NOEC (EC0) was determined to be 50.0 mg/L, the 48h-EC50 was calculated to be 125.5 mg/L and the 48h-EC100 was determined to be >100.0 mg/L.

The second study was performed in order to evaluate the toxic potential of the hydrolysis product 2,2-Dimethyl-3-lauroyloxy-propanal (aldehyde component) towards freshwater shrimp, using the species Daphnia magna. Six experiments were performed. As the test item is poorly water soluble, the “water-accommodated fraction” was tested in the first experiment. This was done by weighing the nominal load 100 mg/L, adding the corresponding amount of dilution water and shaking vigorously for 24 hours. The resulting solution was filtrated through 0.45 μm filters. The daphnia were immobilized on the surface already after 24 hours caused by an oily phase of the test item. The second experiment was performed likewise but with stirring instead of shaking. The resulting solution was left to stand for about 30 minutes. The lower phase was used for the test which was performed as a semi-static test. After 24 hours, the daphnia were immobilized on the surface. The oily film was visible again. All following experiments were performed under semi-static conditions with medium renewal after 24 hours. For the third and fourth experiment, a stock solution in acetone was prepared. The treatments 0.1 and 1 mg/L were prepared by spiking the corresponding amount of dilution water with this stock solution. The treatment 1 mg/L was used as lying well above maximal water solubility of 0.227 mg/L. No daphnia were immobilized after 48 hours. In the control, the same concentration of acetone didn’t show any effect. Based on these experiments, the fifth experiment was performed as a limit-test using the treatment 1 mg/L with a acetone-spiked test solution. After 48 hours 30 % of the daphnia were immobilized on the surface. Therefore the last experiment was performed using five concentrations between 0.1 and 1 mg/L. Twenty daphnia were exposed to the test item for 48 hours in a semi-static test system. After 24 and 48 hours, the immobilized daphnia were counted. As no daphnia were dead after 24 hours, but immobilized on the surface, some of them were able to remobilize during the test. This was potentially due to the test item’s physicochemical properties causing film-formation at the top of the water phase (as noted at higher concentrations). None of the animals were immobilized in the control. The 48 h EC50 was determined about 1 mg/L (exact value after liner extrapolation: 1.02 mg/L) and the 48 h NOEC was 0.18 mg/L. These data were already submitted in a NONS dossier under Directive 92/32/EEC (notification number 05 -04 -1922 -00 from 2005 -10 -25) and considered valid by the German Competent Authority (BAUA).

The acute toxicity of the second hydrolysis product 2-(2-aminoethoxy)ethanol to Daphnia magna was studied under static conditions over a period of 48 hours according to OECD TGD 201 and EU-method C.2. Six concentrations ranging from 31.25 to 500 mg/L were tested. Immobilisation was observed. The following endpoint was derived: EC50 (48 h) > 500 mg/L; EC0 (48 h) = 125 mg/L (see ECHA dissemination homepage).

In addition data from the analogues substance Sika Hardener LI (Dodecanoic acid , 3-[[3-[[[2,2-dimethyl-3-[(1-oxododecylo)oxy]propylidene]amino]methyl]-3,5,5-trimethylcyclohexyl]imino]-2,2-dimethylpropyl ester, CAS 932742-30-8) is reported. Sika Hardener LG and Sika Hardener LI both have the same aldehyde component (2,2-Dimethyl-3-lauroyloxy-propanal), but different amine components. Data from the OECD toolbox support the use of data from Sika Hardener LI also as read across fro Sika Hardener LG (see attachment in section 13). In this study, young Daphnia were exposed in a semi-static test to the test item for 48 hours. A supersaturated test item solution was prepared by dispersing the test item amount (nominal load of 100 mg/L) into the test medium two days before the start of the renewal periods (on day -2 and -1). This solution was shaken for 24 hours at app. 30 °C, then was left settling for 24 hours at app. 20 °C and thereafter filtrated through a 0.45 μm filter. The nominal concentration tested was 100.0 mg/L. In this 48-hour semi-static acute toxicity test with Daphnia magna the 48 h EC50 value was >100 mg/L, and the 48 h NOEC was determined to be 100 mg/L. The results were based on the nominal concentrations. Due to the fast hydrolysis and the very low water solubility of the test item this approach was done. Analytical measurements were not possible due to fast hydrolysis.

Conclusion:

Due to the physical-chemical properties of Sika Hardener LG (fast hydrolysis) it is not possible to perform standard ecotoxicology tests. It was decided to do the test by preparing a WAF and base the EC50 and NOEC calculations on nominal concentrations (justification see study record). A value of 125.5 mg/L was derived. The data presented for the supporting substance Sika Hardener LI with a LC50 of 100 mg/L well supports the results obtained for Sika Hardener LG. Results from the two hydrolysis products further support the results and lead to same environmental classification and labelling (aq. chronic cat.3). For 2,2-Dimethyl-3-lauroyloxy-propanal (aldehyde component) different tests were done to assess the ecotoxicology towards daphnia. In the end it was also not possible to use measured data for EC50 calculations (details see above). As the substance is readily biodegradable the LC50 of 1.02 mg/L does not lead to a classification. The data with 2-(2-aminoethoxy)ethanol with an LC50 of > 500 mg/L show that the amine component does not represent a danger for environment. These data confirm that the classification and labelling and the hazard assessment of the ecotoxicology of Sika Hardener LG can be based on the main study with the product itself.