Amides, C12-18 and C18-unsatd., N,N-bis(hydroxyethyl)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

From February 15, 1996 to April. 13, 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

KL 2 due to RA and no analytical dose verification.

Justification for type of information:

Refer to the section 13 for details on the category justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Direct dispersion in water
- Solvent control: Dechlorinated tap water
- Chemical name of vehicle (organic solvent): Water
- Concentration of vehicle in test medium (stock solution): 500 mg/L
- Preparation of test solution: Test material (500 mg) was dispersed in 1 L of dechlorinated tap water to give stock solution from which aliquots (40, 72, 128, 224 and 400 mL) were separately dispersed in 20 L (final volume) of dechlorinated tap water to give the test series 1.0, 1.8, 3.2, 5.6 and 10 mg/L.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Donnington Fish Farm, Upper Swell, Gloucestershire, UK
- Age at study initiation (mean and range, SD): Juvenile
- Length at study initiation (length definition, mean, range and SD): 5.0 cm (SD = 0.4)
- Weight at study initiation (mean and range, SD): 1.57 g (SD = 0.33)
- Feeding during test: No


ACCLIMATION
- Acclimation period: 8 d
- Acclimation conditions (same as test or not): Maintained in a glass fiber tank with a single pass water renewal system
- Type and amount of food: Commercial trout pellets, discontinued 24 h prior to definitive study

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

100 mg/L as CaCO3

Test temperature:

14 ± 1°C

pH:

7.6-7.9

Dissolved oxygen:

≥ 9.1 mg O2/L

Salinity:

No data

Nominal and measured concentrations:

Nominal concentrations:
- Range finding study: 1.0, 10 and 100 mg/L
- Definitive study: 1.0, 1.8, 3.2, 5.6 and 10 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): Closed
- Material, size, headspace, fill volume: 20 L glass exposure vessel
- Aeration: Aerated via narrow bore glass tubes
- Renewal rate of test solution (frequency/flow rate): Daily
- No. of organisms per vessel: Three in range finding study, ten in definitive study
- No. of vessels per concentration (replicates): Two
- No. of vessels per vehicle control (replicates): Two
- Biomass loading rate: 0.79 g bw/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated tap water


OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h darkness


EFFECT PARAMETERS MEASURED (with observation intervals if applicable): 3, 6, 24, 48, 72 & 96 h post-exposure


TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: Not applicable
- Range finding study
- Test concentrations: 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: Results from range-finding study showed no mortalities at 1 mg/L. However, mortalities were observed at 10 & 100 mg/L.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 2.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 1.8 - 3.2

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: -

Details on results:

- Behavioural abnormalities: Behavioural responses (increased pigmentation, fish swimming at the bottom of the test vessels, loss of equilibrium and the presence of moribund fish) to exposure were observed at test concentration ≥ 1.8 mg/L.
- Mortality of control: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

No data.

Sublethal observations / clinical signs:

Table 1: Cumulative mortality data in the range-finding study

Nominal concentration (mg/L)	Cumulative Mortality (Initial Population: 3)
	24 h	48 h	72 h	96 h
Control	0	0	0	0
1.0	0	0	0	0
10	3	3	3	3
100	3	3	3	3

Table 2: Cumulative mortality data in the definitive study

Nominal concentration (mg/L)	Cumulative Mortality (Initial Population: 10)						% mortality
	3 h	6 h	24 h	48 h	72 h	96 h	96 h
Control	0	0	0	0	0	0	0
1.0	0	0	0	0	0	0	0
1.8	0	0	0	0	0	0	0
3.2	0	0	0	0	5	10	100
5.6	0	0	2	10	10	10	100
10	0	0	10	10	10	10	100

* R1-R2 = Replicates 1 & 2

Table 3: Behavioural responses to exposure in the definitive study

Nominal concentration (mg/L)	Behavioural Responses*	Time (h)
		3	6	24	48	72	96
Control	No abnormalities detected
1	No abnormalities detected
1.8	Increased pigmentation					2/10
	Swimming at bottom with
	increased pigmentation						4/10
3.2	Swimming at bottom with
	increased pigmentation				8/10	2/5	A/D
	Moribund				2/10	3/5
5.6	Swimming at bottom		5/10	8/8	A/D
	Increased pigmentation			8/8
10	Swimming at bottom	10/10	5/10	A/D
	Loss of equilibrium		5/10

* Unless stated no adverse reactions were observed.

A/D = All fish dead

Table 4: Results based on the mortality data in the definitive study

Time (h)	LC50 (mg/L)	95% Confidence Limits (mg/L)
3	>10	-
6	>10	-
24	6.7	5.8 -7.8
48	4.2	3.2 -5.6*
72	3.2	2.6 -3.8
96	2.4	1.8 -3.2*

* Concentrations resulting in 0 and 100 % mortality respectively

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the nominal 96 h LC50 and NOEC were determined to be 2.4 and 1.0 mg/L, respectively.

Executive summary:

A study was conducted to evaluate the acute toxicity of the read across substance, C8-18 and C18-unsatd. DEA, to Rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203, in compliance with GLP. Based on the results of a range-finding study, groups of 20 trout were exposed to the test substance (directly dispersed in water) at nominal concentrations of 1.0, 1.8, 3.2, 5.6 and 10 mg/L for 96 h under semi-static conditions. Mortality and adverse reactions were determined at 3, 6, 24, 48, 72 and 96 h. No analytical dose verification was conducted. Mortality occurred as of 72 h at 3.2 mg/L and as of 24 h at the two higher doses. Behavioural responses (increased pigmentation, fish swimming at the bottom of the test vessels, loss of equilibrium and presence of moribund fish) were observed as of 1.8 mg/L. Under the study conditions, the nominal 96 h LC50 and NOEC were determined to be 2.4 and 1.0 mg/L, respectively (Wetton, 1996).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

From November 07, 1994 to November 17, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

KL 2 due to RA

Justification for type of information:

Refer to the section 13 for details on the category justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

1. Oxygen conc. was 35-57% of the ASV in the first 24 h. 2. No analytical monitoring was carried out for the test material

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

The test solution was prepared daily in > 90% aerated dilution water. The test material was found to be soluble up to 1000 mg/L in water.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The fish were received from the import quarantine and further quarantined for 14 days before start of the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No

Hardness:

250+/-25 mg CaCO3/L

Test temperature:

23+/-1°C

pH:

7.8+/-0.2

Dissolved oxygen:

35-57% of the air saturation value after the first 24 h; 51-89% of the ASV from 48-96 h.

Salinity:

Not reported

Nominal and measured concentrations:

Nominal conc.: 1, 2, 4, 6 and 8 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 5 L glass vessels
- Aeration: The dilution water of the test was aerated exceeding 90% of the air saturation value.
- Renewal rate of test solution (frequency/flow rate): Every 24 hours
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): One
- No. of vessels per control (replicates): One
- No. of vessels per vehicle control (replicates): One
- Biomass loading rate: 0.6 g fish/L


Dilution water composition:
Source-Reverse osmosis water.

CaCl2- 11.76 mg/mL (1 L solution)
MgSO4- 4.93 mg/mL (1 L solution)
NaHCO3- 2.59 mg/mL (1 L solution)
KCl- 0.23 mg/mL (1 L solution)

All four solutions were mixed and diluted to 40 L. Prior to use, the pH was adjusted and then aerated with an air pump.


OTHER TEST CONDITIONS
- Adjustment of pH: With HCl or NaOH
- Photoperiod: 16/8 light:dark
- Light intensity: Not specified, normal laboratory illumination


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality at start, 3, 24, 48, 72 and 96 h intervals


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 4.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality at test conc.-100% mortality was observed at the highest conc. by 24 h and at 6 mg/L by 96 h. There was one mortality at 4 mg/L by 96 h and no mortality at the two lower doses.
- Behavioural abnormalities: Not observed
- Observations on body length and weight: Not observed
- Other biological observations: None reported
- Mortality of control: None died
- Other adverse effects control: None
- Abnormal responses: None

Validity criteria were fulfilled in the following manner:
1. Throughout the test, constant conditions were claimed to be maintained
2. There was no mortality in control, hence mortality of control did not exceed 10%
3. Dissolved oxygen concentration was at least 60% of the ASV till the end of the study (however, during first 24 h, the oxygen conc. was 35-57% of the ASV).

Sublethal observations / clinical signs:

None.

Validity criteria fulfilled:

yes

Remarks:

with exceptions that i) the oxygen conc. was 35-57% (below 60%) only in the first 24 h. However, this deviation probably had no effect on the 100% mortality observed at 8 mg/L, ii) no analytical verification of the test material conc. was carried out

Conclusions:

Under the conditions of the study, the nominal 96 h LC50 and NOEC were calculated to be 4.9 and 2.0 mg/L, respectively.

Executive summary:

A study was conducted to determine the acute toxicity of the read across substance, C8-18 and C18-unsatd. DEA, to Zebrafish (Danio rerio) according to OECD Guideline 203, in compliance with GLP. Groups of 10 fish were exposed to the test substance (directly dissolved in water) at nominal concentrations of 0, 1, 2, 4, 6 and 8 mg/L for 96 h under semi-static conditions. Mortality was observed at 0, 3, 24, 48, 72 and 96 h. No analytical dose verification was conducted. Under the conditions of the study, the nominal 96 h LC50 and NOEC were calculated to be 4.9 and 2.0 mg/L, respectively (Olsson, 1994).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

2.4 mg/L

Additional information

A study was conducted to evaluate the acute toxicity of the read across substance, C8-18 and C18-unsatd. DEA, to Rainbow trout (Oncorhynchus mykiss) according to OECD Guideline 203, in compliance with GLP. Based on the results of a range-finding study, groups of 20 trout were exposed to the test substance (directly dispersed in water) at nominal concentrations of 1.0, 1.8, 3.2, 5.6 and 10 mg/L for 96 h under semi-static conditions. Mortality and adverse reactions were determined at 3, 6, 24, 48, 72 and 96 h. No analytical dose verification was conducted. Mortality occurred as of 72 h at 3.2 mg/L and as of 24 h at the two higher doses. Behavioural responses (increased pigmentation, fish swimming at the bottom of the test vessels, loss of equilibrium and presence of moribund fish) were observed as of 1.8 mg/L. Under the study conditions, the nominal 96 h LC50 and NOEC were determined to be 2.4 and 1.0 mg/L, respectively (Wetton, 1996).

A study was conducted to determine the acute toxicity of the read across substance, C8-18 and C18-unsatd. DEA, to Zebrafish (Danio rerio) according to OECD Guideline 203, in compliance with GLP. Groups of 10 fish were exposed to the test substance (directly dissolved in water) at nominal concentrations of 0, 1, 2, 4, 6 and 8 mg/L for 96 h under semi-static conditions. Mortality was observed at 0, 3, 24, 48, 72 and 96 h. No analytical dose verification was conducted. Under the conditions of the study, the nominal 96 h LC50 and NOEC were calculated to be 4.9 and 2.0 mg/L, respectively (Olsson, 1994).