Butanedioic acid, sulfo-, 4-[1-methyl-2-[(1-oxo-9-octadecenyl)amino]ethyl] ester, disodium salt

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:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control, 11.0, 14.0, 17.0, 20.0, 23.0 mg/L test item equivalent to 0, 3.97, 5.05, 6.14, 7.22, 8.30 mg/L a.s.
- Sample storage conditions before analysis: Specimens were stabilized with 50% methanol. All specimens were stored deep frozen (≤ -18 °C) until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item is emulsifiable in water. Therefore; a stock solution was prepared by adding the test item to test medium. The stock solutions were stirred for 10 minutes on a magnetic stirrer and were whitish and turbid.
Stock solution and test vessels were prepared in the following way:
- A stock solution was prepared by weighing of
460.1 mg test item at 0 hours = test start
459.7 mg test item at 24 hours
460.3 mg test item at 48 hours
460.2 mg test item at 72 hours
into a graduated flask and filling up to 1000 mL using test medium (= stock solution A, corresponding to 460.1 mg/L at test start, 459.7 mg/L for renewal at 24 after hours, 460.3 mg/L at 48 h and 460.2 mg/L at 72 hours).
The corresponding amounts of stock solution A were filled with test medium up to 1000 mL to prepare stock solutions B-F. Each stock solution was then mixed with 3 L test medium in the pool.
The test solutions of the chosen concentrations were prepared directly in the test vessels immediately before the transfer of the fish into the test vessels.


- Controls: The control vessels contained test medium (dilution water) only. One day before the start of the test, the test medium was prepared and aerated for 2 hours.

- Evidence of undissolved material (e.g. precipitate, surface film, etc.): no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish – Danio rerio HAMILTON (1892)

- Source: purchased from fish supplier „Zierfischcenter Scalar“ Paul-Geißler-Straße 14, 04808 Wurzen, Germany held in the Test Facility under standardised laboratory conditions


- Length at study initiation (length definition, mean, range and SD): 1.7 +/- 0.2 cm

- Weight at study initiation (mean and range, SD): 0.11 +/- 0.02 g
- Maintenance of the brood fish:

ACCLIMATION
- Acclimation period: 11 days
- Acclimation conditions (same as test or not): Test fish were in good health and were acclimatised in test medium of the same quality used in the test.
- Type and amount of food during acclimation:
- Feeding frequency during acclimation: daily (fish food), until 24 hours before test start,
- Health during acclimation (any mortality observed): During the acclimatisation period, fish were not exposed to the test item. Mortality of fish during the acclimatisation period was 0%.


FEEDING DURING TEST
- Food type: not fed during the test
- Amount: not fed during the test
- Frequency: not fed during the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

230 mg/L CaCO3

Test temperature:

23.9 – 24.3 °C (recorded continuously)pH

pH:

7.73 to 7.96

Dissolved oxygen:

8.44 to 8.67 mg/L

Nominal and measured concentrations:

Nominal Control, 11.0, 14.0, 17.0, 20.0, 23.0 mg/L test item equivalent to 0, 3.97, 5.05, 6.14, 7.22, 8.30 mg/L a.s.
Geometric mean of measured concentrations over 0-96 hours: Control, 10.3, 13.4, 14.1, 20.5, 23.3 mg/L test item equivalent to 0, 3.71, 4.83, 5.11, 7.42, 8.40 mg/L a.s.

Details on test conditions:

TEST SYSTEM
- Test vessel: stainless steel container (approx. 6 L volume)
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 4 L
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.18 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dilution water: reconstituted water according to ISO 6341, prepared 1 day before test start; conductivity of deionised water: ≤ 10 μS/cm (measured: 1.9 μS/cm)
- Culture medium different from test medium: no
- Intervals of water quality measurement: before the start of the test and before each renewal:

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 light/dark phases
- Light intensity: on average 20 μE *m-2*s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The mortality was recorded at 3, 6, 24, 48, 72 and 96 hours. Fish were considered dead if there was no visible movement (e.g. gill movements) and if touching of the caudal peduncle produced no reaction. Dead fish were removed when observed.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.7 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: The fish in the control group and in the treated test concentrations showed no abnormalities during the test or before they died.
- Mortality of control: none
- Other adverse effects control: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

Step-down Cochran-Armitage Test Procedure and Fisher`s Exact Binomial Test with Bonferroni Correction for statistical significance of mortality (p ≤ 0.05, one-sided)
Probit analysis for calculation of the ECx for mortality (with 95% confidence limits)
Statistical program: ToxRat Professional Version 3.3 (Ratte, 20.10.2018)

Validity criteria fulfilled:

yes

Conclusions:

96h- LC50: 5.7 mg a.s./L

Executive summary:

In the Klimisch 1 GLP study from Juckeland (2020) the acute toxicity of C18 unsaturated fatty acids, reaction products with 1-aminopropan-2-ol, maleic anhydride and sodium bisulfite to zebra fish (Danio rerio) was determined in a 96‑hour semi-static test with daily test medium renewal according to the OECD Guideline for Testing of Chemicals No. 203 (1992).

The following nominal concentrations were tested: Control, 11.0, 14.0, 17.0, 20.0, 23.0 mg/L test item equivalent to 0, 3.97, 5.05, 6.14, 7.22, 8.30 mg/L a.s..

The geometric mean of measured concentrations over 0-96 hours were: control, 10.3, 13.4, 14.1, 20.5, 23.3 mg/L test item equivalent to 0, 3.71, 4.83, 5.11, 7.42, 8.40 mg/L a.s.

 

The biological results were based on geometric mean of measured concentrations.

In the blank control and up to 4.83 mg a.s./L all fish survived until the end of the test and no visible abnormalities were observed. At 5.11 mg a.s./L 42.9% of the fish died. At higher concentrations, all fish died. The 96-hour LC50 was calculated to be 5.7 mg a.s./L.

  

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

 

Description of key information

The 96-hour LC50 was calculated to be 5.7 mg a.s./L.

  

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

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

5.7 mg/L

Additional information

In the Klimisch 1 GLP study from Juckeland (2020) the acute toxicity of C18 unsaturated fatty acids, reaction products with 1-aminopropan-2-ol, maleic anhydride and sodium bisulfite to zebra fish (Danio rerio) was determined in a 96‑hour semi-static test with daily test medium renewal according to the OECD Guideline for Testing of Chemicals No. 203 (1992).

The following nominal concentrations were tested: Control, 11.0, 14.0, 17.0, 20.0, 23.0 mg/L test item equivalent to 0, 3.97, 5.05, 6.14, 7.22, 8.30 mg/L a.s..

The geometric mean of measured concentrations over 0-96 hours were: control, 10.3, 13.4, 14.1, 20.5, 23.3 mg/L test item equivalent to 0, 3.71, 4.83, 5.11, 7.42, 8.40 mg/L a.s.

 

The biological results were based on geometric mean of measured concentrations.

In the blank control and up to 4.83 mg a.s./L all fish survived until the end of the test and no visible abnormalities were observed. At 5.11 mg a.s./L 42.9% of the fish died. At higher concentrations, all fish died. The 96-hour LC50 was calculated to be 5.7 mg a.s./L.

  

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