Poly[oxy(methyl-1,2-ethanediyl)], .alpha.-[2-[[2,2-dimethyl-3-[(1-oxododecyl)oxy]propylidene]amino]methylethyl]-.omega.-[2-[[2,2-dimethyl-3-[(1-oxododecyl)oxy]propylidene]amino]methylethoxy]-

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:

2005-03-09 to 2005-06-17

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:

April 13th 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

July 31st 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Remarks:

Analysis of substance concentration technically not feasible because of the rapid hydrolysis of the test item and its degradation products.

Vehicle:

yes

Details on test solutions:

As the test item is poorly water soluble a "water accommodated fraction" ( WAF) was prepared by weighing of the nominal load 100 mg/L, adding the corresponding amount of dilution water and stirring for 24 hours instead of shaking. The resulting solution was left to stand for 15 minutes, then the lower phase was taken for the test.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain: Berlin
- Source: Umweltbundesamt Berin; bred in the LAUS GmbH throughout the year
- Age at study initiation (mean and range, SD): 0 - 24 h
- Feeding during test: No

ACCLIMATION
- Acclimation period: 30 min
- Acclimation conditions: same as test
- Type and amount of food: green algae
- Health during acclimation: No mortality observed during acclimatisation.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

2.502 mmol/L

Test temperature:

20 ± 2°C

pH:

7.8 - 8.4

Dissolved oxygen:

> 7.4 mg/L

Nominal and measured concentrations:

Nominal loading rates used for the WAF:
Test 1: 0, 100 mg/L (test item was shaken during preparation)
Test 2: 0, 100 mg/L (test item was stirred during preparation)
Test 3: 0, 22, 46, 100, 220, 460 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: glass, nominal volume 50 mL, tall shape
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: drinking waterwith an enrichment of certain minerals (as demanded in the guidelines)
- Total organic carbon: 0.82 mg/L
- Pesticides: <0.1 µg/L
- Chlorine: 12.5 mg/L
- Conductivity: 249 µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement:

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

EFFECT PARAMETERS MEASURED: Check of Immobility after 24h and 48h.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 5 concentrations in a geometric series
- Preparation finding study
- Test concentrations: in both (test 1 and 2) a concentration of 0, 100 mg/L was used.
- Results used to determine the conditions for the definitive study:
Test 1: Immobilization 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.
Test 2: The daphnia were immobilized 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 the results, the third experiment was performed under the same conditions with five concentrations in a geometric series.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

350 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 460 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EL100

Effect conc.:

> 460 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

NOELR

Effect conc.:

460 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 460 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

As no analytical method exists for the test item and because of the immediate hydrolysis of the test item, it was planned to analyse the product of hydrolysis 2,2-Dimethyl-3-lauroyloxy-propanal. But this product is not stable as well. As it can be seen from the tests with TOC-determination, the soluble part is too low to be determined as TOC. Therefore no analytical determination of the test item in the test solutions was possible and the biological results were based on the nominal concentrations.

Two concentrations (220, 460 mg/L) showed toxicity between 0 and 100% immobilization. No animals were immobilized in the control.

Because of the immediate hydrolysis, no water solubility could be determined. The calculated value (following EPA-calculation) is much smaller than 1 mg/L. The water solubility of the product of hydrolysis is below 1 mg/L. Therefore it can be stated that the determined effects are definitely above the limit of water solubility and the "worst case" was tested. The effects were probably caused by physical effects of the unsolved parts of the test item.

Results with reference substance (positive control):

Biological results of the reference item including validity criteria for the reference substance:

• The 24h-EC50 of K2C2O7 should lie between 0.6 and 2.1 mg/L.
The 24h-EC50 of K2Cr2O7 was determined as 1.9 mg/L.
• Immobilization in the controls may not exceed 10 %.
Immobilization 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 8.2 mg/L.

All validity criteria are met.

Table 1: Immobilities in first experiment

Nominal Concentration in mg/L	Immobility 24 hours			Immobility 48 hours
	abs.		in %	abs.		in %
0	0/5	0/5	0	-	-	-
100	5/5	5/5	100	-	-	-

Table 2: pH and O2-values first experiment

Nominal Concentration in mg/L	pH		O2 concentration in mg/L
	0 h	24 h	0 h	24 h
0	7.8	8.2	9.1	8.4
100	7.9	8.2	8.8	8.4

Table 3: Mortalities in second experiment

Nominal Concentration in mg/L	Immobility 24 hours			Immobility 48 hours
	abs.		in %	abs.		in %
0	0/5	0/5	0	0/5	0/5	0
100	3/5	0/5	30	0/5	0/5	0

Table 4: pH- and O2 values for second experiment

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	old	old	new	old
0	7.8	8.0	7.8	8.0	8.8	8.2	8.0	8.0
100	7.8	8.1	8.2	8.0	7.9	8.0	8.1	8.2

Table 5: TOC values in mg/L measured during the second experiment

Nominal concentration in mg/L	Test solution used at time	TOC concentrationin mg/L
		30 min. stirring	24 hours stirring
100	0 h	0.09	1.31
100	24 h	1.41	4.72

Table 6: Immobilities during the third experiment

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
22	0/5	0/5	0/5	0/5	0	0/5	0/5	0/5	1/5	5
46	0/5	0/5	0/5	0/5	0	0/5	0/5	0/5	0/5	0
100	0/5	0/5	0/5	0/5	0	0/5	0/5	0/5	0/5	0
220	0/5	0/5	0/5	0/5	0	3/5	3/5	1/5	2/5	45
460	0/5	0/5	0/5	0/5	0	3/5	1/5	4/5	3/5	55

Table 7: pH- and O2-values for the third experiment

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	old	old	new	old
0	7.8	8.1	7.8	8.1	8.0	8.3	7.9	7.8
22	8.3	8.1	7.9	8.2	7.8	8.1	7.8	8.1
46	8.3	8.1	7.8	8.2	7.7	8.3	7.6	8.2
100	8.3	8.1	7.8	8.2	7.7	8.2	7.4	8.1
220	8.4	8.1	7.9	8.2	7.8	8.2	7.4	8.0
460	8.4	8.1	8.0	8.2	7.4	8.3	7.5	8.0

Validity criteria fulfilled:

yes

Conclusions:

Sika Härter LJ was tested for toxicity to aquatic freshwater invertebrates in a 48h-semi-static test according to EU method C.2 revealing an EL50 of 350 mg/L and a NOELR of 100 mg/L.

Executive summary:

Sika Härter LJ was assessed in a GLP-compliant short term toxicity study to aquatic invertebrates according to EU method C.2 and OECD guideline 202. Three experiments were performed. 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. Ten daphnia were exposed to the test item for 48 hours in a static test system. All daphnia were immobilized on the surface after 24 hours. Immediately after the beginning of the test, an oily film on the surface was observed, which was assumed to be responsible for the immobilization, and potentially due to hydrolysis.

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 instead of shaking. The resulting solution was left to stand for 15 minutes, then the lower phase was taken for the test. Ten daphnia were exposed to the test item for 48 hours in a semi-static test system with medium renewal after 24 hours. After 24 hours, 30 % of the daphnia were immobilized on the surface, after 48 hours none of the daphnia were immobilized. The TOC of the test solution was measured after 30 minutes of stirring and after the 24 hour

stirring period. The TOC lay in a range between 0.1 and 4.7 mg/L. Based on the results of these experiments, a third experiment was performed in the same fashion as the second experiment using five concentrations between 22 and 460 mg/L.

Twenty daphnia were exposed to the test item for 48 hours in a semi-static test system with medium renewal after 24 hours. After 24 and 48 hours, the immobilized daphnia were counted. Two concentrations showed toxicity between 0 and 100% immobilization. No animals were immobilized in the control. As no analytical method exists for the test item and because of the immediate hydrolysis of the test item, it was planned to analyse the product of hydrolysis 2,2-Dimethyl-3-lauroyloxypropanal. But this product is not stable as well. As it can be seen from the tests with TOC-determination, the soluble part is too low to be determined as TOC. Therefore no analytical determination of the test item in the test solutions was possible and the biological results were based on the nominal concentrations. Two concentrations (220, 460 mg/L) showed toxicity between 0 and 100% immobilization. No animals were immobilized in the control. The 48h EL50 was determined at 350 mg/L and the 48h NOEC at 100 mg/L.

Because of the immediate hydrolysis, no water solubility could be determined. The calculated value (following EPA-calculation) is much smaller than 1 mg/L. The water solubility of the product of hydrolysis is below 1 mg/L. Therefore it can be stated that the determined effects are definitely above the limit of water solubility and the “worst case” was tested. The effects were probably caused by physical effects of the unsolved parts of the test item. The study is considered valid.

Description of key information

Sika Härter LJ was tested for toxicity to aquatic freshwater invertebrates in a 48h-semi-static test according to EU method C.2 revealing an EC50 of 350 mg/L and a NOEC of 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

350 mg/L

Additional information

Sika Härter LJ was assessed in a short term toxicity study to aquatic invertebrates according to EU method C.2 and OECD guideline 202. Twenty daphnia were exposed to the test item for 48 hours in a semi-static test system with medium renewal after 24 hours. After 24 and 48 hours, the immobilized daphnia were counted. Two concentrations (220, 460 mg/L) showed toxicity between 0 and 100% immobilization. No animals were immobilized in the control. The 48h EC50 was determined at 350 mg/L and the 48h NOEC at 100 mg/L.