C18 unsaturated fatty acids, reaction products with 1-aminopropan-2-ol, maleic anhydride and sodium bisulfite

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sample storage conditions before analysis: Specimens were diluted with methanol (1:1) and were stored deep frozen (≤ -18°C) until they were analysed. Concentrations of the test item C18 unsaturated fatty acids, reaction products with 1- aminopropan-2-ol, maleic anhydride and sodium bisulfite in test solutions were analyzed at test start (0 hours), and at the test end (48 hours) and after the test solution renewal in the ‘fresh’ solutions and in the ‘spent’ test solutions.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution was prepared by adding the test item to test medium. The stock solution was stirred for 10 minutes on a magnetic stirrer.
Stock solution and test vessels were prepared in the following way: - A stock solution was prepared by weighing of 53.0 mg test item (0 and 24 hours) into a graduated flask and filling up to 1000 mL using test medium (= stock solution A, corresponding to a concentration of 53.0 mg/L test item). The test solutions of the chosen concentrations were prepared directly in the test vessels by the dilution of the stock solution A with the test medium immediately before the transfer of the Daphnia 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 overnight.

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

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna STRAUS
- Age at study initiation (mean and range, SD): < 24 hours
- Method of breeding: breeding was performed in glass beakers (50 mL) under the same conditions as in the test; test animals were separated from the adults by means of a pipette. The parental daphniods were held and bred in the test facility under standardised laboratory conditions, test animals derived from a healthy stock, daphnids used for testing were not not first brood progeny.
- Source: RWTH Aachen University, Institute for Environmental Research (Biology V), Worringerweg 1, 52074 Aachen, Germany

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.6 – 20.7 °C (recorded continuously)

pH:

8.27 to 9.01

Dissolved oxygen:

7.58 - 8.04 mg/L

Nominal and measured concentrations:

Nominal control 4.77, 8.59, 15.4, 27.8, 50.0 mg/L test item (spacing factor: 1.8)
The calculated endpoints are based on the nominal concentrations for the test item since the measured concentrations were within 80 to 120% of nominal.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker (25 mL)
- Material, size, headspace, fill volume: 10 mL
- Renewal rate of test solution (frequency): renewal every 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 + 3 additional vessels (1 for measuring of water parameters, 1 for analysis, 1 retain specimen)
- No. of vessels per control (replicates): 4 + 3 additional vessels (1 for measuring of water parameters, 1 for analysis, 1 retain specimen)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water according to ISO 6341
- Culture medium different from test medium: No
- Intervals of water quality measurement: every 24 hours

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

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

Reference substance (positive control):

yes

Remarks:

The reference item potassium dichromate was tested in a separate study to verify the sensitivity of the test system.

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

7.32 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: No abnormal behaviour or appearance was observed (e.g. trapping at surface of water or discoloration).
- Mortality of control: no immobilization in the control
- Other adverse effects control: none
- Abnormal responses: 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:

The determination of the NOEC / LOEC for immobility was carried out using Step-down Cochran-Armitage Test Procedure at 24 and 48 hours after test start. The EC10, EC20 and EC50 values were calculated by Probit analysis according to the maximum likelihood method. During the probit analysis the logarithm of the test concentrations was used to present the ECx results in the figure.
Statistical analysis was performed using the software ToxRat Professional (Version 3.3, Ratte, 20.10.2018)

Validity criteria fulfilled:

yes

Conclusions:

48h-EC50: 7.32 mg a.s../L (Daphnia magna)

Executive summary:

In the Klimisch 1 GLP study from Juckeland (2019) the acute toxicity of C18 unsaturated fatty acids, reaction products with 1-aminopropan-2-ol, maleic anhydride and sodium bisulfite on Daphnia magna was determined in an 48 hour semi-static test according to OECD 202. The test was performed with 4.77, 8.59, 15.4, 27.8, 50.0 mg test item/L and a blank control. Four replicates with 5 daphnids each were set up. Additional replicates were set-up for analytical purposes. After 48 hours 0, 0, 5. 0, 65, 100 and 100% immobilization was observed in the blank control and at nominal test concentrations of control, 4.77, 8.59, 15.4, 27.8, 50.0 mg test item/L, respectively.

The calculated endpoints are based on the nominal concentrations for the test item since the measured concentrations were within 80 to 120% of nominal.

The 48 hour-EC50 was calculated to be 7.32 mg a.s./L All validity criteria were met.

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

Description of key information

The 48 hour-EC50 for Daphnia magna was calculated to be 7.32 mg a.s./L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

7.32 mg/L

Additional information

In the Klimisch 1 GLP study from Juckeland (2019) the acute toxicity of C18 unsaturated fatty acids, reaction products with 1-aminopropan-2-ol, maleic anhydride and sodium bisulfite on Daphnia magna was determined in an 48 hour semi-static test according to OECD 202. The test was performed with4.77, 8.59, 15.4, 27.8, 50.0 mg test item/L and a blank control. Four replicates with 5 daphnids each were set up. Additional replicates were set-up for analytical purposes. After 48 hours 0, 0, 5. 0, 65, 100 and 100% immobilization was observed in the blank control and at nominal test concentrations of control,4.77, 8.59, 15.4, 27.8, 50.0mg test item/L, respectively.

The calculated endpoints are based on the nominal concentrations for the test item since the measured concentrations were within 80 to 120% of nominal.

The 48 hour-EC50 was calculated to be 7.32 mg a.s./L All validity criteria were met.

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