Poly[oxy(methyl-1,2-ethanediyl)], .alpha.-hydro-.omega.-[2-[[2,2-dimethyl-3-[(1-oxododecyl)oxy]propylidene]amino]methylethoxy]-, ether with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol (3:1)

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004-10-06 to 2005-08-17

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:

adopted 2004-04-13

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

cited as 92/69/EWG method C.2 as of 1992-07-31

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

As the test item is poorly water soluble, the "water-accommodated fraction" (WAF) was used in the first experiment. This was done by weighing of 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.

In the second experiment, the WAF was prepared by weighing of the nominal load 100 mg/L, adding the corresponding amount of dilution water and stirring for 24 hours in¬stead of shaking. The resulting solution was left to stand for 15 minutes, then the lower phase was taken for the test. The TOC of the test solution was measured after 30 minutes of stirring and after the stirring period of 24 hours. The TOC lay in a range between 0 and 2.2 mg/L.

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
- Food type: green algae (Desmodesmus subspicatus)
- Feeding during test. none
- Lighting: none

24 hours before the start of the test, the adult animals were separated from the young. 23 hours later, the adults were caught with the help of a glass tube, and the newborn daph-nia, aged between 0 and 23 hours, were sieved from the medium and immediately placed into a 250 mL-beaker containing dilution water. After a settling-in period of 30 minutes, animals which showed no apparent damage were used for the test.

Daphnia magna is bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. They multiply by parthenogenesis, thus being genetically identical. The holding is performed similar to the method described in EN ISO 6341.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg/L

Test temperature:

21.2 - 22.0 °C

pH:

7.8 to 8.2

Dissolved oxygen:

The concentration of dissolved oxygen stayed above 7.6 mg/L throughout the test.

Nominal and measured concentrations:

As solubility lies below 100 mg/L, the water-accommodated fraction was prepared for the test. This was done by weighing of 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 filter.
4.6 / 10 / 22 / 46 / 100 / 220 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers, nominal volume 2 L; The beakers used are made of glass. They were cleaned by scrubbing with detergent, multiple rinsing with drinking water, rinsing with diluted hydrochloric acid and finally rinsing with deionized water. They were then dried at 105 °C.
- Renewal rate of test solution : after 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

The water-accommodated fraction was prepared for the test. This was done by weighing of each nominal load, adding the corresponding amount of dilution water and stirring slowly for 24 hours. The solutions were left to stand for about 15 minutes. Then the lower phase was used for the test.

Preparation of the Test Solutions
A stock solution containing 250 mg K2Cr2O7 /L in deionized water was diluted with dilution water to give a series of concentrations ranging from 0.625 to 3.0 mg/L.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (CAS 7778-50-9)

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

52 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 13 - 320 mg/L

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

4.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

4.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

First Experiment
Immobilities
In the control, none of the daphnia died or showed any signs of abnormal behaviour throughout the test. As all daphnia in the treatment were immobilised on the surface after 24 hours, the test was terminated.

pH and O2, Temperature
The pH values in the test media and the control ranged from 7.8 to 8.2. The concentration of dissolved oxygen stayed above 8.4 mg/L throughout the test. Temperature range was 19.8 - 20.7 °C.

Assessment
Immobilisation of the daphnia was probably caused by the oily film which could be seen 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 unsolved 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.

Second Experiment
Immobilities
In the control, none of the daphnia died or showed any signs of abnormal behaviour throughout the test.

pH and O2 , Temperature
The pH values in the test media and the control ranged from 7.8 to 8.2. The concentration of dissolved oxygen stayed above 8.0 mg/L throughout the test. Temperature range was 20.9 - 21.5 °C.

TOC-Determination
The TOC of the test solutions was measured 30 minutes after begin of the stirring period and after 24 hours of stirring. Th TOC concentration was determined to be 2.2 mg/L after 24h.

The daphnia were immobilised caused by a thin oily film on the surface. Concerning the observed immobility values, a difference could be noticed between 24 and 48 hours. After 24 hours the medium was renewed. Based on these results, the third experiment was performed under the same conditions with six concentrations in a geometric series.

Third Experiment
Immobilities
In the control, none of the daphnia died or showed any signs of abnormal behaviour throughout the test.

pH and O2, Temperature
The pH values in the test media and the control ranged from 7.8 to 8.2. The concentration of dissolved oxygen stayed above 7.6 mg/L throughout the test. Temperature range was 21.2 - 22.0 °C.

Assessment
In the treatments, a thin oily film on the surface could be observed. This film was probably responsible for the immobilisation. As a dose-response correlation is visible, this test was used to determine the biological results.

Results with reference substance (positive control):

The pH values in the test media and the control ranged from 7.9 to 8.2. The concentration of dissolved oxygen stayed above 8.1 mg/L throughout the test. Temperature range was 21.0 - 21.1 °C.
The 24h-EC50 of K2Cr2O7 should lie between 0.6 and 2.1 mg/L. The 24h-EC50i of K2Cr2O7 was determined as 1.9 mg/L.
Immobilisation in the controls may not exceed 10 %. Immobilisation in the controls was 0 %
The concentration of dissolved oxygen at the end of the test must be at least 3 mg/L . The lowest concentration of dissolved oxygen at the end of the test was 7.6 mg/L.
Thus, all validity criteria were met.

Reported statistics and error estimates:

The estimation of the 24h EC50 of the test item was accomplished using the software Origin.The data were evaluated using sigmoidal fit. Since there were only two values between 0 % and 100 % immobility, a linear fit with a probability-y-axis could not be performed.

Immobilities

Nominal Concentration in mg/L	Immobility 24 hours				Immobility 48 hours
	abs.			in %	abs.			in %
0	0/5 0/5	0/5	0/5	0	0/5 0/5	0/5	0/5	0
4.6	0/5 0/5	0/5	0/5	0	0/5 0/5	0/5	1/5	5
10	1/5 1/5	1/5	2/5	25	5/5 5/5	5/5	5/5	100
22	1/5 0/5	2/5	0/5	15	2/5 3/5	3/5	0/5	40
46	2/5 2/5	3/5	2/5	45	3/5 3/5	5/5	2/5	65
100	4/5 3/5	5/5	3/5	75	5/5 5/5	5/5	5/5	100
220	5/5 4/5	4/5	3/5	80	5/5 4/5	5/5	5/5	95

 

pH and O2, Temperature

 

Nominal Concentration in mg/L	pH				O2-Concentration in mg/L
	0 h	24 h		48 h	0 h	24 h		48 h
	new	old	new	old	new	old	new	old
4.6	8.1	8.1	7.8	8.1	7.9	8.1	7.9	8.0
10	8.0	8.1	7.8	8.2	7.7	8.1	7.8	8.0
22	8.0	8.1	7.9	8.2	7.7	8.1	7.6	8.0
46	7.9	8.1	7.9	8.2	7.6	8.1	7.7	8.0
100	7.9	8.2	7.8	8.2	7.6	8.1	7.8	8.0
220	7.9	8.1	7.9	8.2	7.6	8.1	7.6	7.7

 

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (48 h) of Sika Hardener LTJ was calculated to be 27 mg/L. The highest concentration causing no immobility (no observed effect concentration, NOEC) after 48 hours amounted to 4.6 mg/L. The EC100 was 100 mg/L, which was observed after 48 hours.

Executive summary:

The acute toxicity of the Water Accommodated Fraction (WAF) of Sika Hardener LTJ to Daphnia magna (water flea) was tested under semi-static conditions for 48 hours in accordance with OECD guideline no. 202 and EU method C.2. The test item is immiscible with water and therefore a WAF was prepared. Due to the hydrolysis of SIKA Hardener LTJ no analytial method for the determination of the concentration was available. Thus, the nominal concentrations was used for calculation. The EC50 (48 h) was calculated to be 27 mg/L. The highest concentration causing no immobility (no observed effect concentration, NOEC) after 48 hours amounted to 4.6 mg/L. The EC100 was 100 mg/L, which was observed after 48 hours. All validation criteria were fullfilled.

Description of key information

The acute toxicity of the test substance to Daphnia magna was tested under semi-static conditions according to OECD guideline 202. The EC50 (48 h) was calculated to be 27 mg/L. The NOEC was calculated to be 4.6 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

27 mg/L

Additional information

The acute toxicity of the Water Accommodated Fraction (WAF) of Sika Hardener LTJ to Daphnia magna (water flea) was tested under semi-static conditions for 48 hours in accordance with OECD guideline no. 202 and EU method C.2. The test item is immiscible with water and therefore a WAF was prepared. Due to the hydrolysis of SIKA Hardener LTJ no analytial method for the determination of the concentration was available. Thus, the nominal concentrations was used for calculation. The EC50 (48 h) was calculated to be 27 mg/L. The highest concentration causing no immobility (no observed effect concentration, NOEC) after 48 hours amounted to 4.6 mg/L. The EC100 was 100 mg/L, which was observed after 48 hours. All validation criteria were fullfilled.