Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The results refer to technical product as received from the sponsor. The test is performed according to standard guidelines, under GLP, no chemical analyses. IPA is used as solvent, because the test substance is poorly soluble in water. One fish died in the control with IPA, this is not considered to have an impact on the test results, because in the two lowest test concentrations no fish died during the test. In one test vessel the oxygen concentration was just below 60% instead of >60%, this did not have an impact on the test results either, because no fish died in that aquarium and it happened only at 96h. The effects of the test compound seem normal.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

and EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Stock solutions were prepared in IPA due to the poor water solubility of the test substance. The test concentrations were prepared by addition of the respective amounts of stock solution in DSW.
- Controls: One control and one control with 100 uL IPA/L
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Isopropylalcohol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): the highest test concentration (0.22 mg/L) contained 100 ul IPA/L, therefore one control + 100 uL IPA/L was added.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): -

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Strain: -
- Source: a well-known local aquarium retailer
- Age at study initiation (mean and range, SD): -
- Length at study initiation (length definition, mean, range and SD): 2.8 cm approx.
- Weight at study initiation (mean and range, SD): 0.3 g approx.
- Method of breeding: -
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: Before being used in the test, the fish were kept for at least 12 days in the laboratory
- Acclimation conditions (same as test or not): Holding water: Dutch Standard Water
- Type and amount of food: -
- Feeding frequency: 1 to 3 times a day during 6 days a week. Last feeding took place about 24 hours before the start of the test.
- Health during acclimation (any mortality observed): -

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

The fish were kept for at least 12 days in the laboratory to determine the acceptability of the batch.

Hardness:

13 degree dH

Test temperature:

22.4 - 23.0 degree C

pH:

7.4 - 8.2

Dissolved oxygen:

8.2 - 8.3 mg O2/l at the start of the test
4.9 - 5.9 mg O2/l at the end of the test
At 0.069 mg/L the oxygen concentration went just below 60% of the saturation value at the end of the test. This is not considered to have an impact on the outcome of the test because no fish died at this test concentration.

Nominal and measured concentrations:

0.047, 0.069, 0.10, 0.15 and 0.22 mg/l (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open; covered with a glass plate
- Material, size, headspace, fill volume: 5 L glass aquaria
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): solutions were renewed after 48 hours
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.7 g/L approx.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Synthetic water (Dutch Standard Water)
- Total organic carbon: -
- Particulate matter: -
- Metals: -
- Pesticides: -
- Chlorine: -
- Alkalinity: -
- Ca/mg ratio: 100 mg/L NaHCO3, 20 mg/L KHCO3. 200 mg/L CaCl2*2H2O, 180 mg/L MgSO4*7H2O
- Conductivity: -
- Culture medium different from test medium: No
- Intervals of water quality measurement: -


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 12 hours
- Light intensity: -


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality, behaviour, appearance


TEST CONCENTRATIONS
- Range finding study: Four range finding studies were performed
- Test concentrations:
First: 0.01, 0.1, 1.0 and 10 mg/L
Second: 0.022 and 0.046 mg/L
Third: 0.04, 0.05, 0.063, 0.079 and 0.1 mg/L
Fourth: 0.03, 0.07, 0.12 and 0.18 mg/L
- Results used to determine the conditions for the definitive study:
First: No mortality at 0.01 mg/L and all fish died within 4 hours at 0.1, 1.0 and 10 mg/L. This result was doubtful because very low amounts of a highly concentrated stock were used and the possibility of pipetting errors could not be ruled out.
Second: No mortality was observed at both concentrations within 92 hours.
Third: Only at 0.1 mg/L 70% of the fish died within 96 hours, in the other concentrations no mortality was observed.
Fourth: 100% mortality at 0.12 mg/L after 98 hours and at 0.18 mg/L after 17 hours. No mortality at 0,03 and 0.07 mg/L.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% Confidence limits 0.09 - 0.12 mg/L

Details on results:

- Behavioural abnormalities: At 0.10 mg/L one fish was floating upside down from 76 to 96 hours.
- Mortality of control: No mortaility in the control. In the control + IPA one fish was found dead after 44 hours.
- Other adverse effects control: No
- Abnormal responses: No

Results with reference substance (positive control):

- Results with reference substance valid?: yes
- Mortality: No mortality at 100 mg/L and the control, one fish died in 29 hours at 210 mg/L and at 460 mg/L all fish were found dead at 21 hours.
- LC50: -
- Other: One fish was floating upside down at 210 mg/L from 53 hours till the end of the test

Reported statistics and error estimates:

The LC50 was determined with an LC50 programme of Grifioen (RIZA) based on a model of Kooyman (1981).

Sublethal observations / clinical signs:

Number of surviving animals

concentration (mg/L)		0h	4h	20h	24h	28h	44h	48h	52h	68h	72h	76h	92h	96h
control		7	7	7	7	7	7	7	7	7	7	7	7	7
control + IPA		7	7	7	7	7	6	6	6	6	6	6	6	6
0.047		7	7	7	7	7	7	7	7	7	7	7	7	7
0.069		7	7	7	7	7	7	7	7	7	7	7	7	7
0.10		7	7	7	7	7	7	7	7	6	5*	5*	5*	5*
0.15		7	7	2	1	1	1	1	1	0
0.22		7	4	0

* one fish was almost dead, flaoting upside down

Validity criteria fulfilled:

yes

Remarks:

The oxygen level in one aquarium went just below 60% of the saturation value, but this is not considered to have an impact on the outcome of the test because no fish died at this concentration.

Conclusions:

LC50 = 0.1 mg/L (nominal)

Executive summary:

N-tallow alkyltrimethylenediamines were tested in an acute toxicity test with Brachydanio rerio (zebra fish) under semi-static conditions in accordance with EEG
method C.1 (updated version 11/1989) and OECD Test Guideline 203 (draft updating 12/1989).
No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations.
The LC50 (96 h) was calculated to be 0.1 mg/L with 95% confidence limits of 0.09 and 0.12 mg/L.
The highest concentration causing no mortality (no observed effect concentration, NOEC) after 96 hours amounted to 0.069 mg/l, whereas 100% mortality was produced within 68 hours at 0.15 mg/L.
Sublethal effects were observed at 0.10 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

supporting study

Study period:

18-04-1990 till 18-05-1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The test was performed in three separate tests with different fish batches. The LC50 is however determined in the last of the three tests in which also the LC0 and LC100 was determined. No quantification of the exposure concentration by chemical analyses. No purity/impurity data of the test substance is available

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

(1984)

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

(1984)

Principles of method if other than guideline:

The test was performed in three steps
First: 1 mg/L and control were tested
Second: 0.1 mg/L and control were tested
Third: 0.22, 0.5 and control were tested

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

The test substance was brought into test water and homogenized using ultra-Turrax and a sonicationbath. The test solutions were homogenized in the test vessels during 2 hours. Test concentrations: 0, 0.1, 0.22, 0.5 and 1 mg/L
At 0.22 and 0. 5 mg/L inhomogenity was observed as deposits at the water surface and bottom of the test vessels. This was not observed at the other test concentrations.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Barchydanio rerio
- Strain: (Hamilton-Buchanan)
- Source: West Aquarium 3422 Bad Lauterberg
- Age at study initiation (mean and range, SD):
Hatching date: 19.10.89/ 01.02.90/07.02.90
Date of receival: 09.01.90/24.04.09/03.05.90
- Length at study initiation (length definition, mean, range and SD):
Batch 1/90/L2 23.04.90 n=10 range: 2.9 - 3.4 average: 3.2 S = 0.16
Batch 4/90/L2 07.05.90 n=10 range: 2.4 - 2.9 average: 2.7 S = 0.18
Batch 5/90/L3 14.05.90 n=10 range: 2.7 - 3.4 average: 3.0 S = 0.24

ACCLIMATION
- Acclimation period: 14 days, 10 days for batch 5/90/L3
- Acclimation conditions (same as test or not): 22 deg C
- Type and amount of food: Tetramin twice a day ad libitum
- Health during acclimation (any mortality observed): No remarkable observations

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

in reconstitued lab wate

Hardness:

according to ISO/DIS 7346/1

Test temperature:

21.2 - 22.5 deg C

pH:

7.6 - 8.3

Dissolved oxygen:

7.1 - 9.2 mg/L

Salinity:

according to ISO/DIS 7346/1

Nominal and measured concentrations:

test concentrations: 0, 0.1, 0.22, 0.5 and 1 mg/L (nominal)

Details on test conditions:

Glass aquaria of L/B/H of 30/22/24 cm with a volume of 10 L. Thermostrated at 22 deg C. 12 hours light/12 hours darkness about 700 Lux. Without aeration. Dilution water according to ISO/DIS 7346/1 pH 8.1 - 8.3

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.148 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

0.22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

At 0.1 and 0.5 mg/L sublethal effects were observed like: reduced activity, change in appearance and breathing rate.
At 1 mg/L none of these effects could be observed because the fish died before the first evaluation moment.
Fish that had died appeared shiny metallic,

Results with reference substance (positive control):

-

Reported statistics and error estimates:

LC50 is determined as geometrical mean between LC0 and LC100 in the absence of partial effects.

Sublethal observations / clinical signs:

48 h       96 h

0        0/30      0/30

0.1      0/10      0/10

0.22     10/10     10/10         

0.5       10/10     10/10

1        10/10     10/10

Validity criteria fulfilled:

no

Remarks:

There is no evidence demonstrating that the test substance concentration has been maintained

Conclusions:

The validity is of the test is hampered by the absence of purity data and the absence of evidence demonstrating that the test substance concentration has been maintained during the test. Despite the fact that the test substance is most likely stable during the test the observation that deposits have been observed at the LC0 and LC100 test concentrations which makes the LC50 less reliable.
The information presented in the report is sufficient and transparent which should allow a repetition of the test
It is not expected that a repetition of the test will lead to significantly different results

Executive summary:

The short -term (96 h) toxicity of Genamin SHP 100 (CAS no: 68603 -64 -5) to Danio rerio was observed to be 0.148 mg/L (LC50 96 h). The test substance is poorly water soluble and therefore mechanical equipment was used to prepare the test solutions. No chemical analyses was performed in the absence of suitable analytical techniques, which means that the effect data are based on nominal concentrations.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-05-01 to 1990-05-18

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No chemical analyses

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

Not Applicable

Vehicle:

no

Details on test solutions:

The test solutions were prepared from stock dispersions of the test substance in deionized water.
The test substance was dispersible in water. The clear dispersion was prepared by adding the test substance to water, heating under stirring to approximately 56°C and subsequently cooling down. A clear, homogenous dispersion was obtained. The test concentrations were prepared by addition of the respective amounts of stock dispersion to the DSW.
The stock dispersion used in the definitive test contained 1g of the test substance /L of deionized water.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The fish were approximately 2.8 cm in size and the medium wetweight per fish was approximately 0.3 g. The fish for the definitive test were obtained from a well-known local aquarium retailer and entered in the in-house purchase register as Batch 17-4-90. Before being used in the definitive test the fish were kept for at least 12 days in the Akzo Research Laboratories, dept. CRL, in conformity with Standard Operation Procedure T42, to determine the acceptability of the batch. The water they were kept in was Dutch Standard Water.
The fish were fed one to three times each day (6 days a week), the last feeding taking place about 24h before the test was started.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

no data

Test temperature:

22-24°C

pH:

7.6-8.1

Dissolved oxygen:

Lowest oxygen concentration measured throughout the test was 78% of the air saturation value.

Salinity:

not relevant

Nominal and measured concentrations:

No chemical analyses of the test substance dissolved in the test media were performed. Therefore all concentrations are nominal concentrations:
0.1-0.15-0.22-0.32-0.46 mg/L. One control was included in the test. 7 fish were used per test concentration and control.

Details on test conditions:

The test was performed as a semi-static test with the solution being renewed after 48hours.
As test vessels were used 5L glass aquaria containing 3L of test medium, which were covered with a glass plate.
The biomass load of the test vessels was approximately 0.7 g of biomass/L. During the test, the test medium was not aerated and the animals were not fed.
The test was carried out in a temperature controlled incubator. During the test the temperature was continuously registered in the liquid of the control vessel by means of a temperature sensor. The light regime was 12hours of ambient light per day, provided by fluorescent tubes. Measurements of pH and the oxygen concentrations were carried out daily.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95%CL: 0.15-0.16 mg/L

Details on results:

The highest concentration causing no mortality (NOEC) after 96h amounted to 0.1 mg/L whereas 100% mortality was produced within 23h with 0.22 and 0.32 mg/L. 0.46 mg/L killed all fish within 4h.
No other effects were observed, except at the concentrations of 0.15 to 0.32 mg/L where some of the fish showed reduced activity and some were floating upside down before they died.

Results with reference substance (positive control):

Potassium dichromate was used in order to check the quality of the batch fish used in the test. The concentrations of reference substance used in this test were 100-210-460 mg/L. The pH of the stock solution was adjusted to approximately 8 with NaOH.
A LC50-48h of 205.5 mg/L was obatined.

Reported statistics and error estimates:

The LC50 was determined with an LC50 program of Griffioen (RIZA) based on a model of Kooyman (1981).

Validity criteria fulfilled:

yes

Remarks:

see details below

Conclusions:

The validity criteria were fulfilled:
- the mortality of the control was <10%
- the oxygen concentration was >60%
- the test was validated using a reference substance
but the concentrations of test substance were not analyzed during the test and thus the results are expressed in terms of nominal concentrations.

Executive summary:

Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates

was tested in an acute toxicity test with Brachydanio rerio under semi-static conditions according to OECD 203 guideline and GLP requirements. The LC50 -96h was calculated to be 0.15 mg/L. No chemical analyses of the test concentrations were performed during the test and thus the results are expressed in terms of nominal concentrations. But all the validity criteria were fulfilled.

Description of key information

The 96h-LC50 of Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates to zebrafish is 0.15 mg/L in an OECD TG 203 test under GLP. 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

0.15 mg/L

Additional information

Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates (CAS 1313206-64-2) was tested in an acute toxicity test with zebrafish under semi-static conditions according to OECD 203 guideline and GLP requirements. The LC50 -96h was calculated to be 0.15 mg/L. No chemical analyses of the test concentrations were performed during the test and thus the results are expressed in terms of nominal concentrations. But all the validity criteria were fulfilled.

 

Tests have been performed with comparable methods on linear alkyl diamines with C12 -18 chains. Similarly to tests performed on diamine acetate, most of these tests were conducted without chemical analyses. The available acute fish data for the diamines category which covers an alkyl chain length range from C12 to C18 reveal a comparable toxicity (0.08 to 0.32 mg/L), independent of the alkyl chain length. Thus the toxicity to fish of diamines and diamines acetates are similar. These results confirm that the toxicity of the diamines acetates is not determined by the presence of acetates and that substances have the same ecotoxicty profile.

For more details, see the read-across justification document ("Justification in support of cross-reading from diamines to diamine acetates").