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

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:

2007-06-05 to 2007-06-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: highest concentration and control
- Sampling Method: 2 mL from the approximate centre of the test vessel at t=0 h, t = 24 h and t = 96 h
- 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):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Length at study initiation (length definition, mean, range and SD): 2.7 ± 0.1 cm
- Weight at study initiation (mean and range, SD): 0.56 ± 0.1 g
- Method of breeding: F1 from a single parent-pair bred in UV-treated water.
- Feeding during test: No feeding from 24 hours prior to the test and during the total test period.

ACCLIMATION
- Acclimation period: At least 12 days
- Acclimation conditions (same as test or not): same
- Type and amount of food: pelleted fish food (Nutra 3.0 T, TROUW NUTRITION, Putten, The Netherlands)
- Feeding frequency: Daily
- Health during acclimation (any mortality observed):In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

180 mg CaCO3 per litre and a pH of 7.7 ± 0.3

Test temperature:

control: 20.7 - 21.4°C
100 mg/L: 20.9 - 21.1°C

pH:

control: 7.4 - 7.6
100 mg/L: 7.2 - 7.5

Dissolved oxygen:

control: 5.9 (aeration introduced on day 2) - 8.9 mg/L
100 mg/L: 6.0 (aeration introduced on day 2) - 8.9 mg/L

Nominal and measured concentrations:

nominal: 0, 0.1, 1.0, 10 and 100 mg/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 96-hour test period (96-98%)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 10 L, all-glass, containing 9 L of test solution.
- Aeration: The test media were not aerated, except for a period of 22.5 hours during nominal days 2 and 3 of the test.
- No. of organisms per vessel: 3 (0.1, 1, 10 mg a.i./L), 7 (control and 100 mg a.i./L)
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: Control and 100 mg a.i./L: 0.44 g fish/L; 0.10, 1.0 and 10 mg a.i./L: 0.19 mg fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands)
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH, nitrate and nitrite concentration and ammonia concentration: once a week. Temperature: daily.
In addition, pH and temperature were measured before transferring the fish to the test system.

OTHER TEST CONDITIONS
- Adjustment of pH: yes, pH 7.7 ± 0.3
- Photoperiod: 16 h/d


EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Mortality and sublethal effects at 4, 24, 48, 72 and 96 hours. In addition, every afternoon from day 0 and every morning from day 1 to observe for any dead or severely distressed fish.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Test concentrations: 0, 0.1, 1, 10 and 100 mg a.i./L

Reference substance (positive control):

yes

Remarks:

pentachlorophenol

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

No mortality or clinical effects were observed at any of the concentrations tested.

Results with reference substance (positive control):

- Results with reference substance valid: Yes
96h LC50 = 0.15 mg/L (95% ci 0.10 - 0.22 mg/L)
range of historical data: 96h LC50 0.10 - 0.46 mg/L

Sublethal observations / clinical signs:

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 96-hour test period (96-98%).

All test conditions remained within the ranges prescribed by the protocol (pH: 6.0-8.5, constant within 1 unit; temperature 20-24°C, constant within 2°C; oxygen > 60% of air saturation). Aeration was introduced after 48 hours of exposure, as the oxygen concentration tended to drop below the optimum level for testing with carp, i.e. below 5 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present test Amphopropionate C8 induced no visible effects in carp at or below 100 mg a.i./L. Hence, the 96 h LC50, exceeded 100 mg/L, based on active ingredient concentrations.

Executive summary:

In a 96 h acute toxicity study according to OECD 203 guideline (1992) and EU Method C.1 (1992), carp (Cyprinus carpio) were exposed to Amphopropionate C8 (50.6% a.i.) at nominal concentrations of 0 (control), 0.1, 1.0, 10 and 100 mg a.i./L under static conditions. 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 96-hour test period (96-98%). 3 test organisms were exposed to 0.1, 1 and 10 mg a.i./L and 7 test organisms to 0 mg a.i./L (control) and 100 mg a.i./L.

The 96 h LC50 was > 100 mg a.i./L.  The EC50 and NOEC values, based on mortality/sublethal effects, were both > 100 mg a.i./L.  No visible sublethal effects were observed under the conditions of the present test. 

Results Synopsis

Test organism size/age: Cyprinus carpio (mean length: 2.7 ± 0.1 cm; mean weight: 0.56 ± 0.1 g)

Test Type: static

LC50:  > 100 mg a.i./L                       

NOEC:  > 100 mg a.i./L                       

Endpoint(s) Effected: mortality and sublethal effects

Description of key information

96 h LC50 > 100 mg a.i./L ( analytically verified nominal concentration); OECD TG 203, Cyprinus carpio, GLP, RL1

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

In a 96 h acute toxicity study according to OECD 203 guideline (1992) and EU Method C.1 (1992), carp (Cyprinus carpio) were exposed to Amphopropionate C8 (50.6% a.i.) at nominal concentrations of 0 (control), 0.1, 1.0, 10 and 100 mg a.i./L under static conditions. 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 96-hour test period (96-98%). 3 test organisms were exposed to 0.1, 1 and 10 mg a.i./L and 7 test organisms to 0 mg a.i./L (control) and 100 mg a.i./L.

The 96 h LC50 was > 100 mg a.i./L.  The EC50 and NOEC values, based on mortality/sublethal effects, were both > 100 mg a.i./L.  No visible sublethal effects were observed under the conditions of the present test.