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 fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

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 203 (Fish, Acute Toxicity Test)

GLP compliance:

no

Analytical monitoring:

not specified

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 causing a dispersion of the test substance. Then, the solution was filtrated (WAF preparation)

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM

- Length at study initiation: average length 2 cm
- Feeding during test: yes
- Frequency: daily


ACCLIMATION
- Acclimation period: minimum 14 days
- Acclimation conditions: same as test conditions
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Total exposure duration:

96 h

Test temperature:

21 +/- 1°C

pH:

pH test solution: 6.7
pH control solution: 6.8

Dissolved oxygen:

Dissolved oxygen concentration 73% of air saturation

Nominal and measured concentrations:

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

Details on test conditions:

TEST SYSTEM
- Test volume: 7L
- No. of organisms per vessel: 7

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: 25 ml each of solution (CaCl2, MgSO4, Na2003 and KCI) were mixed and the total volume made up to 1 litre
with deionised water (according to Annex II OECD 203)

OTHER TEST CONDITIONS
- Photoperiod:light-dark cycle 12 hours
- Light intensity: indirect day-similar light

EFFECT PARAMETERS MEASURED: mortality

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: WAF preparation

Details on results:

No effect was observed under the tested conditions, all fishes were alive at the end of the test (96 h), being the EC50 value >10000 mg/L

Sublethal observations / clinical signs:

Table 1. Percentage dead fish after exposure to the test substance

Concentration of the test substance (mg/L)	Dead fish (%)                      24 h             48 h            72 h             96 h
Control	0	0	0	0
10	0	0	0	0
100	0	0	0	0
1000	0	0	0	0
10000	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

No toxic effects were observed in the test organisms under Licowax C exposure within the range of water solubility

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 Nov - 03 Dec 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity 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: 10 mL test solution was sampled directly from each test vessel at test initiation and after 24 hours.
- 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 24 °C. Test water was filtered with No. 2 filter paper.
- Controls: dilution water
- Evidence of undissolved material (e.g. precipitate, surface film, etc): test water was always colorless observed at 0, 24, 48, 72 and 96 hours after initiation.

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka
- Source: Amishima Fish Farm, Yokohama, Japan
- Age at study initiation: about 6 months
- Length at study initiation (mean and range): 2.17 cm (1.91 - 2.40 cm)
- Weight at study initiation (mean and range): 0.172 g (0.107 - 0.226 g)
- Feeding during test: no
- Food type: Tetramin
- Amount: 2% body weight
- Withdrawal of food: 24 hours before and during exposure

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): similar to test condition: (acclimation condition: flow-through, test condition: semi-static)
- Type and amount of food: Tetramin, 3% body weight
- Health during acclimation (any mortality observed): < 5% motality during 7 days before study initiation

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

52 mg/L (CaCO3)

Test temperature:

23.5 - 24.3 °C (24 ± 1 °C)

pH:

7.0 - 7.6

Dissolved oxygen:

7.0 - 8.0 mg/L (more than 60% of saturated oxgen)

Nominal and measured concentrations:

Nominal concentration: 100 mg/L of test item
Measured concentration (geometric mean): 0.0274 mg/L of test substance

Details on test conditions:

TEST SYSTEM
- Test vessel: 5L aquarium
- Material, fill volume: glass, 5 L
- Aeration: Yes
- Renewal rate of test solution (frequency): once in 24 hours
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: ?

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: charcoal-treated, dechlorinated and aerated tap water in Yokohama, Japan
- Alkalinity: 42 mg/L
- Intervals of water quality measurement: 6 months

OTHER TEST CONDITIONS
- Adjustment of pH: not adujusted
- Photoperiod: 16 hours in the light, 8 hours in the dark
- Light intensity: <1000 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mortality, abnormal behavior, mobility (swim ability) observed after 24, 48, 72 hours exposure and at the end of the study

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: control, 100 mg/L (loading rate)
- Results used to determine the conditions for the definitive study: no mortality and no toxicity was observed after 96 hours

Reference substance (positive control):

yes

Remarks:

copper sulfate pentahydrate (analytical grade)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

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:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.027 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.027 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Remarks:

refers to N N'-ethylene-bis-octadecanamide

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: no abnormality was observed
- Observations on body length and weight: not determined
- Other biological observations: not determined
- Mortality of control: 0%
- Other adverse effects control: no
- Abnormal responses: no
- Effect concentrations exceeding solubility of substance in test medium: not observed

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- LC50: 0.53 mg/L (range: mean ± SD: 0.87 ± 0.28 mg/L)
- Other: This test was performed before this main test.

Reported statistics and error estimates:

no statistics was applied

Sublethal observations / clinical signs:

Table 1. The concentration of the test item in test water (Semi-static Condition)

Loading rate* (mg/L)	Measured concentration (mg/L) (percent of nominal)		Mean measured concentration** (mg/L)
	0 h	24 h
control	<0.0002	<0.0002	-
100	0.0293	0.256	0.0274

* the ratio of test item to water (in mg/L) used in the preparation of the WAF.

** geometric mean

Table 2. Mortality of the Medaka exposued to the test substance

Nominal concentration (mg/L)	Measured concentration (mg/L)	Cumulative mortality
		0 h	24 h	48 h	72 h	96 h
control	-	0	0	0	0	0
100	0.0274	0	0	0	0	0

 

Table 3. Observed toxicological symptoms

Nominal concentration (mdg/L)	Measured concentration (mg/L)	Abnormalities
		0 h	24 h	48 h	72 h	96 h
control	-	N	N	N	N	N
100	0.0274	N	N	N	N	N

N.: no toxicological symptom was observed

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

16 Nov - 18 Nov 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: Circular on Test Methods of New Chemical Substances (Japan), bioaccumulation test

Deviations:

yes

Remarks:

no information of concentration, control and vehicle control

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: 142 ± 2 °C
- Water solubility: < 4.9 μg/L

OTHER PROPERTIES
- Results of test for ready biodegradability: ? which result?
- 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:

not specified

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: After mixing with crystal sugar and HCO-40, it was dissolved in ion-exchanged water
- Controls: not stated
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): crystal sugar and HCO-40
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): not stated

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka
- Source: Nakajima fish farm, Tamana, Japan
- Age at study initiation (mean and range, SD): not stated
- Length at study initiation (mean): 3.4 cm
- Weight at study initiation (mean): 0.33 g
- Feeding during test: not stated

ACCLIMATION
- Acclimation period: > 7 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: not stated
- Pretreatment: Treetment with nitrofran antibiotic (product name: Erubaju, Ueno Fine Chemicals Industry, Osaka, Japan) and saline for 24 hours under a static condition

Test type:

semi-static

Water media type:

freshwater

Total exposure duration:

48 h

Test temperature:

25 ± 2 °C

pH:

at the initiation: 7.9
after exposure: 7.6

Dissolved oxygen:

at the initiation: 7.6 mg/L
after exposure: 6.2 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass container
- Material, fill volume: glass, 4 L
- Aeration: not stated
- Renewal rate of test solution (frequency/flow rate): once in 16 - 18 hours
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): not stated
- No. of vessels per control (replicates): not stated
- No. of vessels per vehicle control (replicates): not stated
- Biomass loading rate: not determined

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ground water
- Intervals of water quality measurement: once in 6 months

OTHER TEST CONDITIONS
- Adjustment of pH: not stated
- Photoperiod: not stated
- Light intensity: not stated

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

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 300 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

No toxicity observed up to the highest tested concentration

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

weight of evidence

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

Reason / purpose for cross-reference:

read-across source

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Conclusions:

Based on the result of three acute toxicity to fish tests it can be concluded that the registered substance exhibited no acute toxicity to fish.

Executive summary:

Two studies investigating the acute toxicity of the Read Across Source substance Amides, C16-C18 (even), N,N'-ethylenebis to two different fish species were available. A report from the Japanese Ministry of Environment (MOE, 2002) was chosen as key study. This test was conducted according to OECD Guideline No. 203: Fish, acute toxicity test (1992), using Oryzias latipes as test organism. The results from this study show that after 96 hours exposure to the test substance, no effects were observed up to the highest attainable concentration (0.027 mg/L, limit of solubility). Therefore, the 96-h LC50 was greater than 0.027 mg/L, and no toxicity within the range of water solubility was reported. The test from the supporting study (Bürger and Heyner, 2002), was carried out according to OECD Guideline No. 203: Fish, acute toxicity test (1992), using Brachydanio rerio (Danio rerio) as test organism, under static conditions. Four nominal test concentrations were used, 10, 100, 1000 and 10000 mg/L, and mortality reported after 24, 48, 72 and 96 hours. At the end of the test (96 hours) no mortality was observed up to the highest concentration tested. Therefore, the 96h EC50 was greater than 10000 mg/L. Additional information was collected from the Japanese Ministry of International Trade and Industry (MITI, 1988), regarding a 48 hour test in which N,N’-ethane-1,2 diyldioctadecanamide did not have an effect on Oryzias latipes, being the 48h-LC50 greater than 300 mg/L (nominal concentration).

Description of key information

Based on the result of three acute toxicity to fish tests it can be concluded that the registered substance exhibited no acute toxicity to fish.

Two studies investigating the acute toxicity of the Read Across Source substance Amides, C16-C18 (even), N,N'-ethylenebis to two different fish species were available. A report from the Japanese Ministry of Environment (MOE, 2002) was chosen as key study. This test was conducted according to OECD Guideline No. 203: Fish, acute toxicity test (1992), using Oryzias latipes as test organism. The results from this study show that after 96 hours exposure to the test substance, no effects were observed up to the highest attainable concentration (0.027 mg/L, limit of solubility). Therefore, the 96-h LC50 was greater than 0.027 mg/L, and no toxicity within the range of water solubility was reported. The test from the supporting study (Bürger and Heyner, 2002), was carried out according to OECD Guideline No. 203: Fish, acute toxicity test (1992), using Brachydanio rerio (Danio rerio) as test organism, under static conditions. Four nominal test concentrations were used, 10, 100, 1000 and 10000 mg/L, and mortality reported after 24, 48, 72 and 96 hours. At the end of the test (96 hours) no mortality was observed up to the highest concentration tested. Therefore, the 96h EC50 was greater than 10000 mg/L. Additional information was collected from the Japanese Ministry of International Trade and Industry (MITI, 1988), regarding a 48 hour test in which N,N’-ethane-1,2 diyldioctadecanamide did not have an effect on Oryzias latipes, being the 48h-LC50 greater than 300 mg/L (nominal concentration).

Additional information

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