[No public or meaningful name is available]

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline compliant GPL study with good documentation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
- Sampling method: For the determination of the actual test item concentrations in this semi-static test, duplicate samples were taken from each treatment just before the start of the first and of the last renewal periods (Days 0 and 3). At the end of these renewal periods (Days 1 and 4), additional samples were taken for determination of the stability of the test item during the renewal periods of 24 hours.

- Sample storage conditions before analysis: All samples were taken from the approximate center of each aquarium without mixing the test medium, and were stored deep-frozen (at about -20 °C) immediately after sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Prior to the start of the test and prior to each test medium renewal, a stock solution of nominal 100 mg/L was freshly prepared by completely dissolving 250 mg of the test item (dosed in the range of 250.02 to 250.7 mg) in 2500 mL of test water using ultrasonic treatment for 15 minutes and stirring for 15 minutes at room temperature. Then, adequate volumes of this intensively stirred stock solution were diluted with test water to obtain the test item concentrations of 29.17 and 9.33 mg/L. These concentrations were further diluted to obtain the test item concentrations of the nominal concentrations of 2.92, 0.93 and 0.29 mg/L. The test media were freshly prepared just before introduction of the fish (= start of the exposure) and before each test medium renewal.

- Controls: dilution water only
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No remarkable observations were made concerning the appearance of the test media. All test media were clear throughout the entire test duration.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Strain: not provided
- Source: The fish were obtained from a breeding culture at Harlan Laboratories.
- Age at study initiation (mean and range, SD): not provided
- Length at study initiation (length definition, mean, range and SD): From the acclimated test fish batch, 10 fish were measured at the start of the test. The mean total body length (measured from the mouth to the end of the tail fin) of the fish was 2.9 ± 0.18 cm (Mean ± SD).
- Weight at study initiation (mean and range, SD): the mean body wet weight was 0.17 ± 0.02 g (Mean ± SD).
- Method of breeding: similar to test conditions
- Feeding during test: no

ACCLIMATION
- Acclimation period: Prior to test start, they were acclimated for one week to the test water and temperature.
- Acclimation conditions (same as test or not): similar
- Type and amount of food: During holding and acclimatization until one day before the start of the test, the fish were fed with a commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-Werke, 49304 Melle / Germany).
- Feeding frequency: not reported
- Health during acclimation (any mortality observed):During holding and acclimatization, no fish died in the test fish batch and all fish were healthy.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

15 mg/L as CaCO3

Test temperature:

20-21 °C

pH:

7.0 to 7.8

Dissolved oxygen:

The oxygen concentration was at least 8.4 mg/L, corresponding to at least 92% oxygen saturation.

Nominal and measured concentrations:

The following nominal concentrations of Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18 unsaturated alkyl) tetrasodium salts were tested: 0.29, 0.93, 2.92, 9.33 and 29.17 mg test item/L corresponding to 0.1, 0.32, 1.0, 3.2 and 10 mg solid content/L (mg s.c./L). Additionally, a control (test water without test item) was tested in parallel. The measured concentrations of the 10 mg s.c/L was within 97 and 100 % of the nominal concentrations. Hence the results were reported based on nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: 5 L glass vessel
- Aeration: not reported, but DO concentration was within the validity range
- Renewal rate of test solution (frequency/flow rate): 24 h renewal interval.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.24 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water (AAP Medium) prepared according to the test guideline was used for this study.
- Culture medium different from test medium: no
- Intervals of water quality measurement: at every renewal of the freshly prepared and aged solutions

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: A 16 hour light to 8-hour dark photoperiod, with a 30-minute transition period was used
- Light intensity: light intensity during the light period was approximately within the range of 140 to 480 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
EC50 96 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.2
- Justification for using less concentrations than requested by guideline: NA
- Range finding study
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: Prior to this full test, a limit test according the Threshold-Approach was conducted with a single concentration of 292 mg test item/L (100 mg solid content/L). This test resulted in 100% mortality within 24 hours

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities:
- Observations on body length and weight: NA
- Other biological observations: In the control and up to and including the highest test concentration 29.17 mg test item/L (10 mg solid content/L), all fish survived until the end of the test and no visible abnormalities were observed.
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No remarkable observations were made concerning the appearance of the test media.
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

The LC50 values at different observation times could not be calculated because up to the highest test concentration no mortality or any kind of symptoms were observed. The NOEC, LOEC, LC0 and LC100 were determined directly from the raw data.

Validity criteria fulfilled:

yes

Executive summary:

In the Klimisch 1 GLP study from Kimmel (2013) theacute toxicity of Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18 unsaturated alkyl) tetrasodium salts to zebra fish (Danio rerio) was determined in a 96‑hour semi-static test with daily test medium renewal according to the EU Commission Directive 92/69/, Part C.1, the Commission Regulation (EC) No 440/2008, Part C.1 and the OECD Guideline for Testing of Chemicals No. 203 (1992). The nominal test item concentrations tested were 0.29, 0.93, 2.92, 9.33 and 29.17 mg test item/L. corresponding to 0.1, 0.32, 1.0, 3.2 and 10 mg solid content/L (mg solid content/L). Additionally, a control group was tested in parallel. In the analyzed test medium samples from the start and end of the first and last test medium renewal periods the measured test item concentrations ranged between 97 and 100% of the nominal value. Consequently, the test water renewal ensured nearly constant concentration of the test item during the test and the reported biological results are related to the nominal test item concentration. In the control and up to and including the highest test concentration 10 mg solid content/L, all fish survived until the end of the test and no visible abnormalities were observed. The 96 hour NOEC and LC₀of the test item to zebra fish were both determined to be 10 mg solid content/L. The 96 hour LC₅₀of the test item was determined to be > 10 mg solid content/L.

The results of this study are considered relevant and reliable for the risk assessment.

 

Description of key information

96 h LC50: > 10 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

10 mg/L

Additional information

In the Klimisch 1 GLP study from Kimmel (2013) theacute toxicity of Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18 unsaturated alkyl) tetrasodium salts to zebra fish (Danio rerio) was determined in a 96‑hour semi-static test with daily test medium renewal according to the EU Commission Directive 92/69/, Part C.1, the Commission Regulation (EC) No 440/2008, Part C.1 and the OECD Guideline for Testing of Chemicals No. 203 (1992). The nominal test item concentrations tested were 0.29, 0.93, 2.92, 9.33 and 29.17 mg test item/L. corresponding to 0.1, 0.32, 1.0, 3.2 and 10 mg solid content/L (mg solid content/L). Additionally, a control group was tested in parallel. In the analyzed test medium samples from the start and end of the first and last test medium renewal periods the measured test item concentrations ranged between 97 and 100% of the nominal value. Consequently, the test water renewal ensured nearly constant concentration of the test item during the test and the reported biological results are related to the nominal test item concentration.In the control and up to and including the highest test concentration 10 mg solid content/L, all fish survived until the end of the test and no visible abnormalities were observed.The 96 hour NOEC and LC₀of the test item to zebra fish were both determined to be 10 mg solid content/L. The 96 hour LC₅₀of the test item was determined to be > 10 mg solid content/L.

The results of this study are considered relevant and reliable for the risk assessment.