Reaction mass of N,N-ethanediylbis- Hexadecanamide, N-[2-[(1-oxohexadecyl)amino]ethyl]-Octadecanamide, Hexadecanoic acid, 12-hydroxystearic acid, reaction products with ethylenediamine, N,N-ethanediylbis-Octadecanamide and Octadecanoic acid, 12-hydroxystearic acid, reaction products with ethylenediamine

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:

experimental study

Adequacy of study:

key study

Study period:

2007

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)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna STRAUS (Clone 5)

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

160 to 180 mg CaCO3/L

Test temperature:

20+/- 2 °C

pH:

7.61 to 7.88

Dissolved oxygen:

8.71 to 9.02

Nominal and measured concentrations:

0 - 100 mg/L (WAF)

Details on test conditions:

ELENDT M4 Culture medium

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: WAF

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: WAF

Basis for effect:

other: Immobilization

Validity criteria fulfilled:

yes

Conclusions:

At the water accommodated fraction (WAF) with the nominal loading level of 100 mg/L no biologically significant effect was determined.

Executive summary:

At the water accommodated fraction (WAF) with the nominal loading level of 100 mg/L no biologically significant effect was determined.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

3 December 2001- 22 January 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a definite amount of the test substance (10 mg; 100 mg; 10 g, etc) was filled into measuring flask and the total volume made up with deionised water at 21 +/- 1°C.Test substance was not soluble in water and Xylol could not be used as solvent due to its toxic properties. The solution was shaken vigorously for 24 h causing a dispersion of the test substance. Then, the solution was filtrated (WAF preparation)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain: Daphnia Magna STRAUS
- Supplier: Umwelt-Bundesamt, Berlin (German Federal Environmental Agency)
- Age at study initiation (mean and range, SD): 6-24 h
- Feeding during test: not fed during test


ACCLIMATION
- Acclimation conditions : same as test
- Type of food: freshwater-algae-suspension
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Test temperature:

20 +/- 1° C

pH:

pH test solution: 6.7
pH control solution: 6.8

Dissolved oxygen:

Dissolved oxygen concentration 73% of the air saturation (reference value > 60%)

Nominal and measured concentrations:

Nominal concentrations : 10, 100, 1000 and 10000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4


OTHER TEST CONDITIONS
- Light: test performance in darkness

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: WAF preparation

Details on results:

After 48 h exposure to the 10000 mg/L test concentration, only 10% immobilised Daphnia were reported

Table 1. Percentage immobile Daphnia after 24 and 48 h exposure to the test substance

Concentration of the test substance (mg/L)	No swimming capability (%) after                   24 h                   48 h
Control	0	5
10	0	0
100	0	0
1000	0	0
10000	10	10

Validity criteria fulfilled:

yes

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 Oct - 28 Oct 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Remarks:

according to the Japanese GLP Standard

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Melting point: 146.6 °C
- Water solubility: < 4.9 μg/L

OTHER PROPERTIES
- Other: solubility: < 200 μg/L in hexane, 57 mg/L in chloroform, < 200 μg/L in ethyl acetate, < 200 μg/L in methanol, < 4.9 mg/L in octanol

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 100 mg/L
- Sampling method: 100 mL test solution was sampled from rest of test water at test initiation and each test vessel after 24 hours respectively.
- Sample storage conditions before analysis: analysis was conducted immediately after sampling, no storage

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: test item is immiscible with water and therefore a WAF was prepared by adding the test item in dilution water. After 30 minutes sonication, test water was stirred (750 rpm) for 24 hours at 20 °C. Test water was filtered with No. 2 filter paper.
- Controls: dilution water

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: Incorporated Administrative Agency, National Institute for Environmental Studies, Tsukuba, Japan
- Age at study initiation: within 24 hours
- Feeding during test: no
- Food type: Chlorella vulgaris
- Amount: 0.2 mg C/day

ACCLIMATION
- Acclimation period: 21 days (2005/10/05 - 2005/10/26)
- Acclimation conditions (same as test or not): similar to test condition (Renewal rate of water and density of animals: acclimation condition: three times in a week and 1 animal/80 mL, test condition: once in 24 hours and 5 animals/100 mL)
- Type and amount of food: Chlorella vulgaris, 0.2 mg C/day
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no mortality, no presence of males and ephippia

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

242 - 246 mg/L (CaCO3)

Test temperature:

20.0 - 20.3 °C (20 ± 1 °C )

pH:

8.1 - 8.3

Dissolved oxygen:

8.3 - 8.8 mg/L (more than 60% of saturated oxygen)

Nominal and measured concentrations:

Nominal concentration: 100 mg/L of test substance
Measured concentration (geometric mean): 0.00223 mg/L of test substance

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL beaker
- Type: closed
- Material, fill volume: glass, 100mL
- Renewal rate of test solution (frequency): once in 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: ?

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium

OTHER TEST CONDITIONS
- Adjustment of pH: not adjusted
- Photoperiod: 16 hours in the light, 8 hours in the dark
- Light intensity: <800 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mobility

TEST CONCENTRATIONS
- Range finding study: performed
- Test concentrations: control, 100 mg/L (loading rate)
- Results used to determine the conditions for the definitive study: immobility at 48 hours: 0%

Reference substance (positive control):

yes

Remarks:

potassium dichromate (reagent grade)

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 39.8 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

refers to N N'-ethylene-bis-octadecanamide

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.002 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.001 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Remarks:

refers to N N'-ethylene-bis-octadecanamide

Basis for effect:

mobility

Details on results:

- Observations on body length and weight: not determined
- Mortality of control: not observed
- Other adverse effects control: not observed

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50: 0.75 ± 0.15 mg/L
- Other: This test was performed every 6 months.

Reported statistics and error estimates:

no statistics was applied

Table 1. Concentration of the test substance in test water (Semi-Static Condition)

Loading Rate (mg/L)	Measured Concentration (mg/L)		Geometric Mean During 24 Hours (mg/L)
	0 Hour new	24 Hours old
Control	<0.00002	<0.00002	-
100	0.00446	0.00112	0.00223

Loading rate: the ratio of test substance to water (in mg/L) used in the preparation of the WAF

new: freshly prepared test solutions

old: test solutions after 24 hours exposure

Table 2. The number of immobilezed Daphnia magna (Percent Immobility)

Loading Rate (mg/L)	Geometric Mean Measured concentration (mg/L)	Cumulative number of immobilized Daphnia (Percent Immobility)
		24 Hour	48 Hours
Control	-	0 (0)	0 (0)
100	0.00223	0 (0)	0 (0)

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Please refer to chapter 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Immobilisation

Validity criteria fulfilled:

yes

Conclusions:

Based on the result of two acute toxicity to daphnia tests it can be concluded that the registered substance exhibited no acute toxicity to Daphnia magna within the range of water solubility.

Executive summary:

Two studies investigating the acute toxicity of the Read Across Source substannce Amides, C16-C18 (even), N,N'-ethylenebis to Daphnia magna were available. A report from the Japanese Ministry of Environment (MOE) was chosen as key study. The test was equivalent to the one included in the OECD Guideline No. 202: Daphnia sp., Part I- Acute Immobilisation Test (1984). The results show that after 48 hours exposure to the test compound, up to the highest attainable concentration (0.0022 mg/L), no immobilization was observed on Daphnia magna. Therefore, the 48h-EC50 was greater than 0.0022 mg/L and no toxicity was observed within the range of water solubility. The test performed in the supporting study (Heyner, 2002) was carried out according to the OECD Guideline No. 202: Daphnia sp., Part I- Acute Immobilisation Test (1984), but not under GLP conditions. Immobilisation was assessed in daphnids exposed to four (nominal) test concentrations, 10, 100, 1000 and 10000 mg/L, after 24 and 48 hours. At the end of the test (48 hours) the 48h-EC50 was greater than 10000 mg/L.

Description of key information

One study investigating the acute toxicity of the registered substance to Daphnia magna is available. The test was carried out according to the OECD Guideline no. 201. At the water-accommodated fraction (WAF) with the nominal loading level of 100 mg/L no biologically significant effect was determined.

 

Two studies investigating the acute toxicity of the Read Across Source substance Amides, C16-C18 (even), N,N'-ethylenebis to Daphnia magna were available. A report from the Japanese Ministry of Environment (MOE) was chosen as key study. The test was equivalent to the one included in the OECD Guideline No. 202: Daphnia sp., Part I- Acute Immobilisation Test (1984). The results show that after 48 hours exposure to the test compound, up to the highest attainable concentration (0.0022 mg/L), no immobilization was observed on Daphnia magna. Therefore, the 48h-EC50 was greater than 0.0022 mg/L and no toxicity was observed within the range of water solubility. The test performed in the supporting study (Heyner, 2002) was carried out according to the OECD Guideline No. 202: Daphnia sp., Part I- Acute Immobilisation Test (1984), but not under GLP conditions. Immobilisation was assessed in daphnids exposed to four (nominal) test concentrations, 10, 100, 1000 and 10000 mg/L, after 24 and 48 hours. At the end of the test (48 hours) the 48h-EC50 was greater than 10000 mg/L.

Based on the result of these three acute toxicity to daphnia tests it can be concluded that the registered substance exhibited no acute toxicity to Daphnia magna within the range of water solubility.