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

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 - 28 January 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study in principle according to guideline and under GLP. No data on actual concentrations but in the algae test from 2018 the test substance was shown to be stable.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

No chemical analysis, water hardness less than adviced

Principles of method if other than guideline:

No measurement of actual concentration, hardness below recommended concentration (50 iso 140 mg/L as CaCO3). 2 reps per group iso 4. Statistics according to Thompson and not Finney.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Direct dispersion in water. 200 mg of test material was dispersed in aged and dechlorinated tap water and the volume adjusted to 2 litres to give a 100 mg/L test concentration from which dilutions are made to give the test series.

Test organisms (species):

Daphnia magna

Details on test organisms:

Name: Daphnia magna Straus
Source: Laboratory culture originating from a strain supplied by the Institut national de Recherche Chimique Appliquée (IRCHA), France
Culture: At 21C in propylene vessels containing two litres of dechlorinated and aged tap water. Cultures were fed daily with a suspension of mixed algae (predominantly Chlorella spp.). Culture conditions ensure that reproduction is by parthogenesis. Gravid adults were isolated 24 hours prior to the initiation of the test. Young daphnids produced overnight were used for testing.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

None

Hardness:

50 mg/L as CaCO3

Test temperature:

22.0 C

pH:

7.5 - 7.7

Dissolved oxygen:

8.0 - 8.7 mgO2/L

Nominal and measured concentrations:

Nominal concentrations:
1.0, 1.8, 3.2, 5.6, 10, 18, 32, 56, 100 mg/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.
Test vessels were glass jars each containing 200 ml test solution and were covered to reduce evaporation. 1 control in duplicate and 9 test concentrations in duplicate. 20 mL test solution per organism, 10 per replicate. No aeration during the test and a 16h light - 8h dark photoperiod. Daphnia were placed in the test solutions after addition of the test material. Daphnia were considered to be immobilised if they were unable to swim for approximately 15 seconds after gentle agitation.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.56 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 1.38 - 1.77 mg/L

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 9.0 - 12 mg/L

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

5.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 4.6 - 5.9 mg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

Concentrations >= 18 mg/L caused complete immobility within 24 hours. No immobilisation was observed up to concentrations of 3.2 mg/L during the study period. At the dose rate of 5.6 mg/L no immobilisation was apparent after 24 hours but 13/20 animals were immobilised after 48 hours. In the 10 mg/L dose group 9/20 animals were immobilised after 24 hours, whilst all animals were immobilised after 48 hours.
The EC50 24h and EC50 48h were determined at 10 and 5.2 mg/L respectively on a nominal basis.
The 48h NOEC was concluded at 3.2 mg/L nominally.

Reported statistics and error estimates:

Analysis of the immobility data by the moving average method of Thompson (1947).

Number of mobile animals

Conc substance	Test vessel
in mg/l	number	0 hours	24 hours	48 hours
control	1	10	10	10
	2	10	10	10
	total	20	20	20
1.0	1	10	10	10
	2	10	10	10
	total	20	20	20
1.8	1	10	10	10
	2	10	10	10
	total	20	20	20
3.2	1	10	10	10
	2	10	10	10
	total	20	20	20
5.6	1	10	10	4
	2	10	10	3
	total	20	20	7
10	1	10	6	0
	2	10	5	0
	total	20	11	0
18	1	10	0	0
	2	10	0	0
	total	20	0	0
32	1	10	0	0
	2	10	0	0
	total	20	0	0
56	1	10	0	0
	2	10	0	0
	total	20	0	0
100	1	10	0	0
	2	10	0	0
	total	20	0	0

Validity criteria fulfilled:

yes

Conclusions:

Unfortunately no preliminary test was done. Instead of nine test concentrations, four would have been sufficient for the same amount of information.
Concentrations >= 18 mg/L caused complete immobility within 24 hours. No immobilisation was observed up to concentrations of 3.2 mg/L during the study period. At the dose rate of 5.6 mg/L no immobilisation was apparent after 24 hours but 13/20 animals were immobilised after 48 hours. In the 10 mg/L dose group 9/20 animals were immobilised after 24 hours, whilst all animals were immobilised after 48 hours. The EC50 24h and EC50 48h were determined at 10 and 5.2 mg/L respectively on a nominal basis. The 48h NOEC was concluded at 3.2 mg/L nominally. Expressed for active ingredient (30%) this would give EC50 values of 3.1, 1.6 and a NOEC of 0.96 mgai/L.

Executive summary:

In this study the acute toxicity of Ampholak YCE to Daphnia magna was studied according to OECD202. The test was performed under GLP as a static test. Each replicate contained 10 daphnids, 2 replicates for control and for the nine test concentrations. Concentrations >= 18 mg/L caused complete immobility within 24 hours. No immobilisation was observed up to concentrations of 3.2 mg/L during the study period. At the dose rate of 5.6 mg/L no immobilisation was apparent after 24 hours but 13/20 animals were immobilised after 48 hours. In the 10 mg/L dose group 9/20 animals were immobilised after 24 hours, whilst all animals were immobilised after 48 hours. The EC50 24h and EC50 48h were determined at 10 and 5.2 mg/L respectively on a nominal basis. The 48h NOEC was concluded at 3.2 mg/L nominally.

Description of key information

Two short-term daphnia studies have been performed with Sodium cocopropylenediamine propionate. 

One acute daphnia study with Sodium cocopropylenediamine propionateis was performed in 1993. The study from Sewell and Grant-Salmon (1993) resulted in a 48h EC50 for daphnia magna of 1.6 mg a.i./L based on nominal test concentrations. The test was performed 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 1993.

In 2022 an additional acute daphnia study with Sodium cocopropylenediamine propionate was performed. The study from Scheerbaum (2022) resulted in a 48 h EC50 of 6.41 mg a.i./L based on nominal test item concentrations. The test was performed under static conditions and the analytically verified measured concentrations of the test item based on the main components C12-amine dipropionate, C14-amine dipropionate, C12-diamine tripropionate and C14-diamine tripropionate at the start of the exposure (0 hours) were in the range of 101% to 119% of the nominal values. At the end of the exposure period (48 hours), the measured concentrations were in the range of 82 to 113% of the nominal values. Therefore, the EC50 value is based on the nominal concentration.

Both acute daphnia tests show similar effect values. Since no analytical measurement was performed in the study from Sewell & Grant-Salmon (1993) the data are less reliable compared to the study from Scheerbaum (2022). Therefore the EC50 of 6.41 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 invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

6.41 mg/L

Additional information