Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite

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:

2013

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The concentrations of the test item were analyzed in one of the duplicate test media samples from the nominal concentrations of 124.5 and 249 mg/L from both sampling times (0 and 48 hours). The samples of the nominal test concentrations of 15.56, 31.16 and 62.25 mg/L were not analyzed since the concentrations were below the 48 hour NOEC determined in this test and, thus, were not relevant for the interpretation of the biological results. From the control, one of the duplicate samples was analyzed per sampling time, too.


- Sampling method: For the determination of the actual test item concentrations, duplicate samples were taken from each treatment at the start and at the end of the test (after 48 hours). For the 48 hour stability samples, additional flasks with adequate volumes of the freshly prepared test media of all test concentrations and the control were incubated during the test period under the same conditions as in the actual test (but without daphnids).

- Sample storage conditions before analysis: All samples were stored deep-frozen (at about -20 °C) immediately after sampling. Based on pre-experiments the test item was found to be stable in the test water under these storage conditions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test medium of the highest nominal concentration of 249 mg/L was prepared by completely dissolving 249.8 mg of test item in 1000 mL of test water under intense stirring for 15 minutes at room temperature. Adequate volumes of this test medium were serially diluted with test water to prepare the lower concentrated test media. The test media were prepared just before introduction of the daphnids (i.e., start and end of the test).
- Controls: dilution water only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): NA
- Concentration of vehicle in test medium (stock solution and final test solution): NA
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The test media containing the nominal concentrations of up to and including 31.16 mg substance/L (12.5 mg solid content/L) were clear solutions throughout the entire test duration. At the highest concentrations of 62.25, 124.5 and 249 mg substance/L (25, 50 and 100 mg solid content/L), the test item was observed at the surface of the test media after the start of the test. After 24 hours all test solutions were clear, however at the end of the test in the test media containing the nominal concentrations of 124.5 and 249 mg substance/L (50 and 100 mg solid content/L) the test item was observed at the surface again and was slightly yellow colored. However, analytical measurements confirmed the correct dosage of the test item and stability in the test media over the test period of 48 hours. The biological results were related to the nominal concentrations of the test item.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna Straus
- Strain: clone 5
- Source: A clone of this species (defined by the supplier as clone 5) was originally supplied by the University of Sheffield / UK in 1992. Since that time, the clone has been bred at Harlan Laboratories in reconstituted water of the quality identical to the water quality used in the tests (in respect to pH, main ions, and total hardness) and under temperature and light conditions identical to those of the test.
- Age at study initiation (mean and range, SD): At the start of the test, the organisms used in the test were 6 - 24 hours old and were not first brood progeny.
- Feeding during test: None
- Food type: NA
- Amount: NA
- Frequency: NA

ACCLIMATION
- Acclimation period: Not needed since the cultures were kept under conditions similar to the testing conditions

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3 (nominal from medium)

Test temperature:

20-21 °C

pH:

6.9 - 7.1

Dissolved oxygen:

dissolved oxygen concentrations in the test media and control were at least 8.1 mg/L

Nominal and measured concentrations:

control and 15.56, 31.16, 62.25, 124.5 and 249 mg/L (corresponding to 6.25, 12.5, 25, 50 and 100 mg solid content/L), measured concentrations were of the test concentrations of 124.5 and 249 mg/L were between 92 and 93% of the nominal values at the start of the test and between 85 and 91% at the end of the test.

Details on test conditions:

TEST SYSTEM
- Test vessel: 100-mL glass beakers filled with 70 mL of test medium
- Type (delete if not applicable): The test vessels were covered with glass plates to reduce the loss of water by evaporation and to avoid the entry of dust into the solutions.
- Material, size, headspace, fill volume: 100-mL glass beakers filled with 70 mL of test medium
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 16 mL/daphnia

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water according to ISO 6341
- Culture medium different from test medium: no
- Intervals of water quality measurement: start and end

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16-hour light to 8-hour dark cycle with a 30-minute transition period
- Light intensity: approximately between 390 and 560 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
immobilization at 0, 24, and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: NA
- Range finding study: not reported
- Test concentrations: not reported
- Results used to determine the conditions for the definitive study: not reported

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

72 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Immobilization

Remarks on result:

other: 95% C.I.: 55 - 83 mg as/L

Details on results:

- Behavioural abnormalities: not reported
- Other biological observations: none
- 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: The test media containing the nominal concentrations of up to and including 31.16 mg substance/L (12.5 mg solid content/L) were clear solutions throughout the entire test duration. At the highest concentrations of 62.25, 124.5 and 249 mg substance/L (25, 50 and 100 mg solid content/L), the test item was observed at the surface of the test media after the start of the test. After 24 hours all test solutions were clear, however at the end of the test in the test media containing the nominal concentrations of 124.5 and 249 mg substance/L (50 and 100 mg solid content/L) the test item was observed at the surface again and was slightly yellow colored. Analytical measurements confirmed the correct dosage and the stability of the test item under test conditions
- Effect concentrations exceeding solubility of substance in test medium: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: as expected
- EC50/LC50: 0.94 mg/L
- Other: For evaluation of the quality of the daphnia clone and the experimental conditions, potassium dichromate is tested as a positive control twice a year. The result of the latest positive control test in September 2012 (48-hour EC50: 0.94 mg/L, Harlan Laboratories Study D64333) showed that the sensitivity of the test organisms was within the internal historical range (48-hour EC50 from 2000 to 2012: 0.43 1.1 mg/L).

Reported statistics and error estimates:

The NOEC, EC0 and EC100 were determined directly from the raw data. The 24- and 48-hour EC50 and the 95% confidence limits were calculated by Moving Average Interpolation

Validity criteria fulfilled:

yes

Conclusions:

The EC50 is 72 mg as/L, the NOEC is 50 mg as/L

Executive summary:

In the Klimisch 1 GLP study from Kimmel (2013) the acute toxicity of Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite on Daphnia magna was determined in an 48 hour static test according to OECD 202 and EU method C.2. The test was performed with concentrations of 15.56, 31.16, 62.25, 124.5 and 249 mg/L (corresponding to 6.25, 12.5, 25, 50 and 100 mg solid content/L) and a blank control. Four replicates with 5 daphnids each were set up. After 48 hours 0, 0, 0. 0, 0 and 95 % immobilization was observed in the control and at test concentrations of 6.25, 12.5, 25, 50 and 100 mg solid content/L, respectively. Analytical measurements confirmed that the test solutions were correctly dosed and stable throughout the exposure period. Therefore the data were evaluated based on nominal concentrations. The EC50 is 72 mg as/L, the NOEC is 50 mg as/L.

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

Description of key information

EC50: 72 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

72 mg/L

Additional information

In the Klimisch 1 GLP study from Kimmel (2013) the acute toxicity of Reaction products of ricinoleic acid with 2-aminoethanol and maleic acid and sodium hydrogensulfite on Daphnia magna was determined in an 48 hour static test according to OECD 202 and EU method C.2. The test was performed with concentrations of 15.56, 31.16, 62.25, 124.5 and 249 mg/L (corresponding to 6.25, 12.5, 25, 50 and 100 mg solid content/L) and a blank control. Four replicates with 5 daphnids each were set up. After 48 hours 0, 0, 0. 0, 0 and 95 % immobilization was observed in the control and at test concentrations of 6.25, 12.5, 25, 50 and 100 mg solid content/L, respectively. Analytical measurements confirmed that the test solutions were correctly dosed and stable throughout the exposure period. Therefore the data were evaluated based on nominal concentrations. The EC50 is 72 mg as/L, the NOEC is 50 mg as/L.

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