N-(2-hydroxyethyl)-N-[2-[(1-oxooctyl)amino]ethyl]-β-alanine

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:

2007-06-05 to 2007-06-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control and 100 mg a.i./L
- Sampling method: Frequency: at t=0 h and t=48 h; Volume: 2 mL from the approximate centre of the test vessels. At the end of the exposure period, the replicates were pooled at each concentration before sampling.
- Sample storage conditions before analysis: Samples were stored in a freezer until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of test solutions started with two solutions of 100 mg a.i./L (198 mg/L, based on formulation) applying 16 minutes of magnetic stirring to accelerate the dissolving of the test substance in the test medium, after which the solutions were pooled. Part of the solution was used as highest test concentration and part was used to prepare the lower test concentrations.
- Controls: Test medium without test substance or other additives
- Evidence of undissolved material (e.g. precipitate, surface film, etc):The final test solutions were all clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: In-house laboratory culture with a known history.
- Age at study initiation (mean and range, SD): For the test selection of young Daphnia with an age of < 24 hours, from parental daphnids of more than two weeks old.
- Method of breeding:
Start of each batch: with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
Maximum age of the cultures: 4 weeks
Renewal of the cultures: Alter 7 days of cultivation half of the medium twice a week.
Temperature of medium: 18-22°C
Feeding: Daily, a suspension of fresh water algae.
Medium: M7, as prescribed by Dr. Elendt-Schneider (Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antenna!
damage in Daphnia magna Straus. Protoplasma 154, 25-33).
- Feeding during test: no feeding
- Validity of batch: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

180 mg/L expressed as CaCO3 and the pH: 7.7 ± 0.3.

Test temperature:

19.7 - 20.9°C

pH:

control: 7.6 (t=0h); 7.7 (t=48h)
100 mg a.i./L: 7.5 (t=0); 7.7 (t=48h)

Dissolved oxygen:

control: 8.9 (t=0h); 8.9 (t=48h)
100 mg a.i./L: 8.9 (t=0); 8.6 (t=48h)

Nominal and measured concentrations:

nominal: 0, 0.1, 10, 100 mg a.i./L
Analysis of the samples taken during the combined limit/range-finding test showed that measured concentrations were stable and in agreement with nominal throughout the 48-hour test period (94-97%).

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 100 mL, all-glass
- Aeration: no aeration of the test solutions
- No. of organisms per vessel: 5 per vessel containing 80 ml of test solution
- No. of vessels per concentration (replicates): 2 vessels per concentration for 0.1, 1 and 10 mg/L , 4 vessels for 100 mg/L
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of test medium: Adjusted ISO medium (M7, as prescribed by Dr. Elendt-Schneider)
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
lmmobility (including mortality): At 24 hours and at 48 hours.
pH and dissolved oxygen: At the beginning and at the end of the test, in the highest concentration and the control.
Temperature of medium: Continuously in a temperature control vessel, beginning at the start of the test.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Justification for using less concentrations than requested by guideline:
- Range finding study: Limit test with 100 mg a.i./L
- Test concentrations: 0, 0.1, 1, 10 and 100 mg a.i./L

Reference substance (positive control):

yes

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

No significant immobilisation was observed in any of the concentrations tested.

Results with reference substance (positive control):

- Results with reference substance valid: Yes (potassium dichromate)
- EC50:
24 h EC50 = 0.58 mg/L (95% ci 0.53 - 0.68 mg/L)
48 h EC50 = 0.37 mg/L (95% ci 0.33 -0.42 mg/L)
results were within the ranges of historical data

Acceptability of the test

1. In the control, no daphnia became immobilised or trapped at the surface of the water.

2. The oxygen concentration at the end of the test was ≥ 3 mg/L in control and test vessels.

Table 1: Acute immobilisation of daphnia after 48 hours

Concentration Amphopropionate C8 (mg a.i./L)	Vessel number	Number Daphnia exposed	Response at 48 h
			Number	Total %
Control	A B C D	5 5 5 5	0 0 0 0	0
0.10	A B	5 5	0 0	0
1.0	A B	5 5	0 0	0
10	A B	5 5	0 0	0
100	A B C D	5 5 5 5	1 0 0 0	5

Validity criteria fulfilled:

yes

Conclusions:

No significant immobilisation was observed in any of the concentrations tested. The 48 h EC50 of Amphopropionate C8 in Daphnia magna was > 100 mg a.i./L (nominal concentration).

Executive summary:

The 48-hr-acute toxicity of Amphopropionate C8 (50.6% a.i.) to Daphnia magna was studied under static conditions accordign to OECD Guideline 202.  Daphnids were exposed to control and test chemical at nominal concentrations of 0 (control), 0.10, 1.0, 10, 100 mg a.i./L for 48 h.  Mortality and immobilisation were observed daily. 

Analysis of the samples taken during the combined limit/range-finding test showed that measured concentrations were stable and in agreement with nominal throughout the 48-hour test period (94-97%). Under the conditions of the present study Amphopropionate did not induce acute immobilisation of Daphnia magna at or below 100 mg a.i./L, after 48 hours of exposure.

 

Results Synopsis

Test Organism Age: Daphnia magna, <24 h

Test Type: Static

 

EC50: > 100 mg a.i./L

Endpoint(s) Effected:  immobilisation

Description of key information

48 h EC50 > 100 mg a.i./L (analytically verified nominal concentration), OECD TG 202, Daphnia magna, static; GLP, RL1

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The 48-hr-acute toxicity of Amphopropionate C8 (50.6% a.i.) to Daphnia magna was studied under static conditions accordign to OECD Guideline 202.  Daphnids were exposed to control and test chemical at nominal concentrations of 0 (control), 0.10, 1.0, 10, 100 mg a.i./L for 48 h.  Mortality and immobilisation were observed daily. 

Analysis of the samples taken during the combined limit/range-finding test showed that measured concentrations were stable and in agreement with nominal throughout the 48-hour test period (94-97%). Under the conditions of the present study Amphopropionate C8 did not induce acute immobilisation of Daphnia magna at or below 100 mg a.i./L, after 48 hours of exposure.