Dodecanoic acid, 1,1'-[1,6-hexanediylbis[nitrilo(2,2-dimethyl-1-propanyl-3-ylidene)]] ester

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:

2004-10-06 and 2005-08-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- First Experiment
On each day of the test, the treatment was prepared by mixing a stock emulsion containing 1000 mg/L test item with dilution water.

- Second Experiment
On each day of the test, the treatment was prepared by mixing a stock emulsion containing 1000 mg/L test item with dilution water. The mixture was stirred for 30 minutes in order to reach a solution. The TOC was measured after 30 minutes of stirring (= time of medium renewal) and after 24 hours of the test.

- Third Experiment
On each day of the test, the WAF was prepared. 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 about 15 minutes. Then the lower phase was used for the test.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Danio rerio
- Strain: HAMILTON-BUCHANAN
- Source: Weiler´s Pet´s best, Turmstr. 6, 67433 Neustadt
- Age at study initiation (mean and range, SD): sexually immature young fish
- Length at study initiation: 2 ± 1 cm
- Feeding during test
- Food type: No details
- Amount: totalling about 1 - 2 % of body weight per day
- Frequency: three times a day

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions: same as test conditions
- Type and amount of food: NA
- Feeding frequency: 3 times per day
- Health during acclimation: mortality rate didn't surpass 5 %

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

94.3 mg CaCO3/L

Test temperature:

22 - 23 °C

pH:

7.9 - 8.2

Dissolved oxygen:

> 8.0 mg/L throughout the test

Nominal and measured concentrations:

Nominal concnetrations: 100 mg/L per treatment.
No measured concentration.

Details on test conditions:

TEST SYSTEM
- Test vessel: aquaria
- Type: open
- Material, fill volume: glass, 3 L
- Aeration: accomplished with glass tubes, frequency of bubbles 1/s
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution: every 24 ± 1 hours
- No. of organisms per vessel: first and second experiment: 3; third experiment: 7
- No. of vessels per concentration: first and second experiment: 1 vessel (3 L test solution), third experiment: 1 vessel (7 L test solution)
- No. of vessels per control: first and second experiment: 1 vessel (3 L dilution water), third experiment: 1 vessel (7 L dilution water)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Good quality drinking water was used as dilution water
- Culture medium different from test medium: yes (culture medium: chlorine-free tap water)
- Intervals of water quality measurement: yearly

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: 12/12

EFFECT PARAMETERS MEASURED:
Observations were made every 24 hours, measuring pH and O2-concentration of the old and new test solutions in each vessel and documenting mortalities or abnormal behaviour.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: none
- Mortality of control: none
- Any observations that might cause a difference between measured and nominal values: As the test item is poorly water soluble, a thin oily film was observed on the surface of the stock emulsion and on the surface of the treatment as well. The fish didn’t show any abnormal behaviour or mortality caused by this effect. Therefore the experiment was repeated in the same fashion using seven fish per vessel. The TOC measurements gave results between 0 and 2.9 mg/L TOC in the test solutions 30 minutes after stirring and after 24 hours test time. This is barely above the limit of detection. Therefore this non-specific analytical method is not sensitive enough for the determination of the content of the test item in the solutions and the biological results were based on the nominal concentrations.
Because of the immediate hydrolysis, no water solubility could be determined. The calculated value (following EPA-Calculation) is < 1 mg/L. The water solubility of the product of hydrolysis is below 1 mg/L. Therefore it can be stated that the tested concentration is above the limit of water solubility and thus the “worst case” was tested.

Sublethal observations / clinical signs:

Biological Results Test Item

The biological results are presented in the following table:

Table: Biological Results

Parameter	Value	95 %-confidence-interval
24h NOEC	100 mg/L	n.d.
24h LC50	> 100 mg/L	n.d.
24h LC100	> 100 mg/L	n.d.
48h NOEC	100 mg/L	n.d.
48h LC50	> 100 mg/L	n.d.
48h LC100	> 100 mg/L	n.d.
72h NOEC	100 mg/L	n.d.
72h LC50	> 100 mg/L	n.d.
72h LC100	> 100 mg/L	n.d.
96h NOEC	100 mg/L	n.d.
96h LC50	> 100 mg/L	n.d.
96h LC100	> 100 mg/L	n.d.

n.d. = not determined

Validity criteria fulfilled:

yes

Conclusions:

Sika Hardener LH was tested for acute toxicity to freshwater fish in a 96 h-static test according to EU method C.1. The test item showed no mortality up to a nominal concentration of 100 mg/L. The 96 h-LC50 was estimated greater than 100 mg/L. The 96 h-NOEC was 100 mg/L.

Executive summary:

SIKA Hardener LH was assessed in a short-term toxicity to fish study according to EU method C.1 and OECD guideline 203. A water accommodated fraction (WAF) of 100 mg/L was prepared and nominal concentrations of up to 100 mg/L tested.

Three experiments were performed. For the first experiment, a stock emulsion containing 1000 mg/L in dilution water was prepared on each day of the test. This emulsion was used to prepare the test solution containing 100 mg/L. Three fish were exposed to the test item for 96 hours in a semi-static test system with medium renewal every 24 hours. No mortality was observed, either in the treatment or in the control. None of the fish showed any signs of morbidity at the end of the test. The pH and the oxygen values of the water were normal. The test item is poorly water soluble and an oily film was observed on the surface of the stock emulsion as well as on the treatment solution. Thus, the concentration of the test item in the treatment was not reproducible.

In the second experiment, a stock emulsion containing 1000 mg/L in dilution water was prepared on each day of the test. This emulsion was used to prepare the test solution containing 100 mg/L. The mixture of emulsion and dilution water was stirred slowly for 30 minutes in order to reach a solution. Three fish were exposed to the test item for 96 hours in a semi-static test system with medium renewal every 24 hours. No mortality was observed, either in the treatment or in the control. None of the fish showed any signs of morbidity at the end of the test. The pH and the oxygen values of the water were normal. An oily film was observed on the surface in this experiment as well. Thus, the concentration of the test item in the treatment was not reproducible. The TOC of the test solution was measured after 30 minutes of stirring and after 24 hour test time.The TOC lay in a range between 0 and 2.9 mg/L.

In the third experiment, for each medium renewal the “water-accommodated fraction” (WAF) was prepared by weighing of the nominal load 100 mg/L, adding the corresponding amount of dilution water and stirring slowly for 24 hours instead of shaking. The resulting solution was left to stand for 15 minutes, and then the lower phase was taken for the test. Seven fish were exposed to the test item for 96 hours in a semi-static test system with medium renewal every 24 hours. No mortality was observed in the treatment and the control. None of the fish showed any signs of morbidity at the end of the test. The pH and the oxygen values of the water were normal. As no analytical method exists for the test item and because of the immediate hydrolysis of the test item, initially the hydrolysis product 2,2-Dimethyl-3-laureoyloxy-propanal should have been analysed.But this product is not stable as well.The soluble part is too small to be determined as TOC (as has been demonstrated in the respective studies “toxicity against alga and daphnia”). Therefore no analytical determination of the test item in the test solutions was possible and the biological results were based on the nominal concentrations. The following results for the test item Sika Härter LH (VP), were determined (species Danio rerio HAMILTON BUCHANAN):

24-h NOEC = 100 mg/L nominal concentration

24-h LC50 > 100 mg/L nominal concentration

48-h NOEC = 100 mg/L nominal concentration

48-h LC50 > 100 mg/L nominal concentration

72-h NOEC = 100 mg/L nominal concentration

72-h LC50 > 100 mg/L nominal concentration

96-h NOEC = 100 mg/L nominal concentration

96-h LC50 > 100 mg/L nominal concentration

 

Because of the immediate hydrolysis, no water solubility could be determined. The calculated value (following EPA-Calculation) is < 1 mg/L. The water solubility of the product of hydrolysis is below 1 mg/L. Therefore it can be stated that the tested concentration is above the limit of water solubility and thus the “worst case” was tested.

Description of key information

SIKA Hardener LH was assessed in a short-term toxicity to fish study according to EU method C.1 and OECD guideline 203. A water accommodated fraction (WAF) of 100 mg/L was prepared and nominal concentrations of up to 100 mg/L tested. Young fish were exposed in a static test to the test item for 96 hours at a nominal concentration of 100.0 mg/L. The 96 h-LC50 was determined to be greater than 100 mg/L and the 96 h-NOEC was determined to be 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

SIKA Hardener LH was assessed in a short-term toxicity to fish study according to EU method C.1 and OECD guideline 203. A water accommodated fraction (WAF) of 100 mg/L was prepared and nominal concentrations of up to 100 mg/L tested.

Three experiments were performed. For the first experiment, a stock emulsion containing 1000 mg/L in dilution water was prepared on each day of the test. This emulsion was used to prepare the test solution containing 100 mg/L. Three fish were exposed to the test item for 96 hours in a semi-static test system with medium renewal every 24 hours. No mortality was observed, either in the treatment or in the control. None of the fish showed any signs of morbidity at the end of the test. The pH and the oxygen values of the water were normal. The test item is poorly water soluble and an oily film was observed on the surface of the stock emulsion as well as on the treatment solution. Thus, the concentration of the test item in the treatment was not reproducible.

In the second experiment, a stock emulsion containing 1000 mg/L in dilution water was prepared on each day of the test. This emulsion was used to prepare the test solution containing 100 mg/L. The mixture of emulsion and dilution water was stirred slowly for 30 minutes in order to reach a solution. Three fish were exposed to the test item for 96 hours in a semi-static test system with medium renewal every 24 hours. No mortality was observed, either in the treatment or in the control. None of the fish showed any signs of morbidity at the end of the test. The pH and the oxygen values of the water were normal. An oily film was observed on the surface in this experiment as well. Thus, the concentration of the test item in the treatment was not reproducible. The TOC of the test solution was measured after 30 minutes of stirring and after 24 hour test time.The TOC lay in a range between 0 and 2.9 mg/L.

In the third experiment, for each medium renewal the “water-accommodated fraction” (WAF) was prepared by weighing of the nominal load 100 mg/L, adding the corresponding amount of dilution water and stirring slowly for 24 hours instead of shaking. The resulting solution was left to stand for 15 minutes, and then the lower phase was taken for the test. Seven fish were exposed to the test item for 96 hours in a semi-static test system with medium renewal every 24 hours. No mortality was observed in the treatment and the control. None of the fish showed any signs of morbidity at the end of the test. The pH and the oxygen values of the water were normal. As no analytical method exists for the test item and because of the immediate hydrolysis of the test item, initially the hydrolysis product 2,2-Dimethyl-3-laureoyloxy-propanal should have been analysed.But this product is not stable as well.The soluble part is too small to be determined as TOC (as has been demonstrated in the respective studies “toxicity against alga and daphnia”). Therefore no analytical determination of the test item in the test solutions was possible and the biological results were based on the nominal concentrations. The following results for the test item Sika Härter LH (VP), were determined (species Danio rerio HAMILTON BUCHANAN):

24-h NOEC = 100 mg/L nominal concentration

24-h LC50 > 100 mg/L nominal concentration

48-h NOEC = 100 mg/L nominal concentration

48-h LC50 > 100 mg/L nominal concentration

72-h NOEC = 100 mg/L nominal concentration

72-h LC50 > 100 mg/L nominal concentration

96-h NOEC = 100 mg/L nominal concentration

96-h LC50 > 100 mg/L nominal concentration

 

Because of the immediate hydrolysis, no water solubility could be determined. The calculated value (following EPA-Calculation) is < 1 mg/L. The water solubility of the product of hydrolysis is below 1 mg/L. Therefore it can be stated that the tested concentration is above the limit of water solubility and thus the “worst case” was tested.