trisodium 3-[{3-[bis(2-carboxylatoethyl)amino]propyl}(C12-18-(even numbered) and C18-(unsaturated) alkyl)amino]propanoate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 October - 1 November 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study according to guideline & GLP. No data on actual concentrations. The algae study from 2018 showed that the test substance is stable during 72 hours. It is reasonable to assume that the test substance remained stable in the fish test as well.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

No chemical analysis

Principles of method if other than guideline:

No measurement of actual concentrations but substance has shown to be stable in the algae test (2018). No preliminary test. Statistics different from guideline.

GLP compliance:

yes (incl. QA statement)

Remarks:

although study director page can not be read

Analytical monitoring:

no

Remarks:

In the algae study from 2018 it was shown that the test substance is stable during 72 hours under algae test conditions. It is reasonable to assume that also in this acute fish study the test substance must have remained stable.

Vehicle:

no

Details on test solutions:

Direct dispersion in water. 0.2, 0.36, 0.64, 1.12 and 2.00 g of test material were each dispensed directly into 20 litres of diluent to give the test series.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

Rainbow trout (Oncorhynchus mykiss)
Source: Parkwood trout farm, Wigmore, kent, United Kingdom.
The stock of fish was held since 17 October 1991 and was acclimatised to test conditions from 17 - 28 October 1991.
The stock fish were maintained in a glass fibre tank with a single-pass water renewal system.
Fish were fed dailey with commercial trout pellets, but feeding was discontinued 24h prior to the test.
Mean standard length = 4.1 cm (SD = 0.3 cm), mean weight = 0.97 g (SD = 0.20 g), no data on age of fish.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

Approximately 50 mg/L as CaCO3

Test temperature:

14.0 °C

pH:

7.4 - 7.6

Dissolved oxygen:

10.0 - 10.2 mg O2/L

Salinity:

Fresh water

Nominal and measured concentrations:

Nominal concentrations:
10, 18, 32, 56, 100 mg/L test substance
3, 5.4, 9.6, 16.8 and 30 mg a.i./L

Details on test conditions:

The test material was suspected to absorb to glassware and so saturation of the absorption sites was achieved by soaking the test vessels overnight prior to the start of the test with the test solutions. At 0 hours the test vessels were emptied, rinsed with the solution to be tested and then refilled with the fresh test solution.
There was a dailey renewal of test media to ensure stability of the test concentrations.

The lightning was controlled at 16h light- 8 h darkness cycle.

Test vessels: Glass aquaria holding 20L of test media. 5 test concentrations plus 1 control (10 animals per test concentration).
Fish were placed at random in prepared test media, loading 0.49 g bodyweight/L.
Aeration via narrow bore glass tubes.

Criteria of death: absence of (i) respiratory movement and (ii) response to a physical stimulation.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 3 - 5.4 mg/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 10 - 18 mg/L

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 10 - 18 mg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

24 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 18 - 32 mg/L

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

28 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 24 - 34 mg/L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

The highest concentration test substance resulting in 0% mortality = 10 mg/L and the lowest test concentration resulting in 100% mortality = 18 mg/L.
At 100 mg/L all fish died within 3 h after exposure. Apart from mortality, only loss of equilibrium was observed at concentrations >= 32 mg/L.
The LC50 (96h) was concluded at 13 mg/L , the NOEC (96h) was concluded at 10 mg/L on a nominal basis.

Results with reference substance (positive control):

n.a.

Reported statistics and error estimates:

Analysis of mortality was carried out by the method of Thompson (1947)

Sublethal observations / clinical signs:

Number of surviving animals

Conc (mg/L)	0h	3h	6h	24h	48h	72h	96h
Control	10	10	10	10	10	10	10
10	10	10	10	10	10	10	10
18	10	10	10	10	10	0	0
32	10	10	10	3	0	0	0
56	10	10	0	0	0	0	0
100	10	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The study was performed according to GLP and guideline. Based on the available data, the highest concentration resulting in 0% mortality = 10 mg/L and the lowest test concentration resulting in 100% mortality = 18 mg/L. At 100 mg/L all fish died within 3 h after exposure. Apart from mortality, only loss of equilibrium was observed at concentrations >= 32 mg/L. The LC50 (96h) was concluded at 13 mg/L , the NOEC (96h) was concluded at 10 mg/L on a nominal basis.

Executive summary:

The acute toxicity of Ampholak YCE to rainbow trout (Oncorhynchus mykiss) was evaluated according to OECD203 and was performed under GLP. It was carried out as a semi-static test, with dose levels of nominally 10, 18, 32, 56 and 100 mg/L. The LC50 (96h) was observed to be 13 mg/L of test substance or 4 mg a.i./L, the NOEC (96h) was concluded at 10 mg/L (or 3 mg a.i./L) on a nominal basis. Loss of equilibrium was observed in animals exposed to concentrations of >= 32 mg/L (or 9.6 mg a.i./L).

Description of key information

Two acute fish studies have been performed with Sodium cocopropylenediamine propionate. 

One acute fish study with Sodium cocopropylenediamine propionate was performed in 1992. The study from Sewell and Wetton (1992) resulted in a 96h LC50 for rainbow trout of 4.00 mg a.i./L based on nominal test concentrations. The test was performed semi-statically and the glassware was presoaked with test solution to prevent any potential sorption to glassware. The test substance concentration was however not quantified as no suitable analytical method was available in 1992.

In 2022 an additional acute fish study with Sodium cocopropylenediamine propionate was performed with a limit test concentration of 10.0 mg test item/L (3.14 mg a.i/L based on 31.4 % active ingredient). The selection of the test concentration was based on the derivation of a threshold concentration (TC) from the data of the test item of results of a daphnia toxicity test (EC 50 (48h) = 20.4 mg test item/L and of an algae toxicity test (ErC50 (0-72 h) > 10.0 mg test item/L) from Scheerbaum (2022) and Klix (2018). The acute fish study from Scheerbaum (2022) resulted in a 96 h LC0 for zebrafish of 3.14 mg a.i./L based on nominal test item concentrations and consequently an LC50 > 3.14. The test was performed under static conditions and the measured concentrations of the test item based on the main components C12-amine dipropionate, C14-amine dipropionate, C12-diamine tripropionate and C14-diamine tripropionate were in the range of 80-120% of nominal. Therefore, the effect concentration is based on nominal value.

Both acute fish tests show similar effect values. Since no analytical measurement was performed in the study from Sewell & Wetton (1992) the data are less reliable compared to the study from Scheerbaum (2022). Therefore the 96 h LC50 of >3.14 mg a.i./L (based on an active ingredient content of 31.4%) will be used as key value for the chemical risk assessment. 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 3.14 mg/L