Amides, C16-C18 (even) , N,N'-ethylenebis

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:

26 - 28 Oct 2005

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)

Deviations:

no

GLP compliance:

yes

Remarks:

according to the Japanese GLP Standard

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
- Intervals of water quality measurement: pH, oxygen content and temperature of test solutions were determined daily

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 after 24 and 48 hours.

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:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL50

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 [%]
		24 Hour	48 Hours
Control	-	0 (0)	0 (0)
100	0.00223	0 (0)	0 (0)

Table 3: Validity criteria for OECD 202.

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	0%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	8.3 - 8.8 mg/L	yes

 

Validity criteria fulfilled:

yes

Remarks:

Please refer to table 3 in section "Any other information on results incl. tables"

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:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

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. 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

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 +/- 1 °C

pH:

pH test solution: 6.7 - 7.2
pH control solution: 6.8

Dissolved oxygen:

Control: 6.5 - 8.1 mg/L
Test solutions: 6.3 - 8.7 mg/L

Nominal and measured concentrations:

Nominal test 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

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: synthetic freshwater
- Intervals of water quality measurement: The pH value and oxygen concentrations were measured at the beginning and the end of the test

OTHER TEST CONDITIONS
- Light: test performance in darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mobility after 24 and 48 hours

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Details on results:

- Mortality of control: 5% after 48 hours
After 48 h exposure to the 10000 mg/L test concentration, only 10% immobilised Daphnia were reported

Results with reference substance (positive control):

Results with Potassium dichromate: EC50 (24 h): 1.0 mg/L

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

Table 2: Validity criteria for OECD 202.

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	5%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	6.3 mg/L	yes

 

Validity criteria fulfilled:

yes

Remarks:

Please refer to table 2 in section "Any other information on results incl. tables"

Description of key information

The substance is not acutely toxic to aquatic invertebrates. 

Additional information

Two studies investigating the acute toxicity of the substance to aquatic invertebrates are available. The key study was conducted according to OECD Guideline 202 (GLP) using Daphnia magna as test organism. The study was conducted as a limit test. A water accommodated fraction (WAF) with a nominal loading rage of 100 mg/L was prepared. The test concentration was verified by LC/MS. Immobilisation of D. magna was not observed. The determined EC50 (48 h) was > 0.00223 mg/L based on the measured concentration (geometric mean). Comparable results were obtained in the supporting study conducted according to OECD 202. Daphnia magna were exposed to nominal test substance loading rates of 10, 100, 1000 and 10000 mg/L (WAF), analytical dose verification was not performed. The determined EC50 (48 h) was >10000 mg/L (nominal, WAF).

Based on the available study results, the substance is not considered acutely toxic to aquatic invertebrates.