Reaction products of alkenes, oxidized, esterified with alkanol

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 August 2015 to 15 September 2015

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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:

RANGE-FINDING TEST
- A sample of each loading rate WAF was taken for chemical analysis at 0 and 48 hours in order to determine the stability of the test item under test conditions.
- All samples were stored frozen prior to analysis.
-Only concentrations within the range to be used for the definitive test were analyzed.

DEFINITIVE TEST
- Samples were taken from the control and 100 mg/L loading rate WAF test group at 0 hours from the bulk test preparation and at 48 hours from the pooled replicates R1 to R4 for quantitative analysis.
- Samples were stored frozen prior to analysis.

Vehicle:

no

Details on test solutions:

TEST WATER
- Reconstituted water (ISO medium) used for both the range-finding and definitive tests is defined in Appendix 4 (attached).

PROCEDURE
- Due to the low solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

VALIDATION OF MIXING PERIOD
- Preliminary work (see Appendix 5, attached) was carried out to determine whether stirring for a prolonged period produced significantly higher measured test concentrations in the WAF.

RANGE-FINDING TEST
- Nominal amounts of test item (5.0, 50 and 500 mg) were each separately added to the surface of 5 L of test water to give the 1.0, 10 and 100 mg/L loading rates respectively.
- After addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface.
- Stirring was stopped after 23 hours and the mixtures were allowed to stand for one hour.
- A wide bore glass tube, covered at one end with Nescofilm, was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel.
- A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal.
- Microscopic inspections of the WAFs showed no micro-dispersions or undissolved test item present.
-The aqueous phase of WAF was removed by mid-depth siphoning (the first 75-100 mL discarded) to give the 1.0, 10 and 100 mg/L loading rate WAFs.
- A dilution of the 1.0 mg/L loading rate was used to produce the 0.10 mg/L loading rate WAF.

DEFINITIVE TEST
- Based on the result of the range-finding test a 'limit test' was conducted at a loading rate of 100 mg/L to confirm that at the highest attainable concentration no immobilisation or adverse reactions to exposure was observed.
- A nominal amount of test item (250 mg) was added to the surface of 2.5 L of test water to give the 100 mg/L loading rate.
- After addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface.
- Stirring was stopped after 23 hours and the mixture allowed to stand for one hour.
- A wide bore glass tube, covered at one end with Nescofilm, was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel.
- A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal.
-The aqueous phase or WAF was removed by mid-depth siphoning (the first 75-100 mL discarded) to give the 100 mg/L loading rate WAF.
- Microscopic inspection of the WAFs showed there to be no micro-dispersions or undissolved test item present.

Test organisms (species):

Daphnia magna

Details on test organisms:

- The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.
- Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium (see Appendix 2, attached) in a temperature controlled room at approximately 19°C.
- The lighting cycle was controlled to give 16 hours light and 8 hours darkness with 20 minute dawn and dusk periods.
- Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin flake food suspension.
- Culture conditions ensured that reproduction was by parthenogenesis.
- Gravid adults were isolated the day before initiation of the test so that young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing.
- The diet and diluent water were considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

None

Hardness:

250 mg/L total theoretical hardness as CaCO3

Test temperature:

22 °C

pH:

8.0 to 8.1

Dissolved oxygen:

8.8 to 8.9 mg O2/L

Salinity:

Not applicable

Nominal and measured concentrations:

RANGE-FINDING TEST
- Nominal loading rates of 0.10, 1.0, 10 and 100 mg/L

DEFINITIVE TEST
- Nominal loading rate of 100 mg/L

Details on test conditions:

RANGE-FINDING TEST
- Daphnids (10) were placed in each test and control vessel and maintained at a temperature controlled room at 21 to 22°C with a photoperiod of 16 hours light and 8 hours darkness for a period of 48 hours with 20 minute dawn and dusk transition periods.
- Each 150 mL test and control vessel contained 100 mL of test media and was covered to reduce evaporation.
- After 24 and 48 hours, the number of immobilised daphnids were recorded.
- The control group was maintained under identical conditions but not exposed to the test item.

DEFINITIVE TEST
- 150 mL glass jars containing approximately 100 mL of test preparation were used.
- At the start of the test daphnids (5) were placed in each test and control vessel at random.
- Four replicate test and control vessels were prepared.
- Test vessels were covered to reduce evaporation and maintained in a temperature controlled room at approximately 22 °C with a photoperiod of 16 hours light (200 to 1200 lux) and 8 hours darkness with 20 minute dawn and dusk transition periods.
- Daphnids were not individually identified, received no food during exposure, and the test vessels were not aerated.
- The control group was maintained under identical conditions but not exposed to the test item.
- The test preparations were not renewed during the exposure period.

Reference substance (positive control):

yes

Remarks:

potassium dichromate conducted between 23 February 2015 and 25 February 2015 (see Appendix 3, attached)

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

RANGE-FINDING TEST
- Cumulative immobilisation data and other observations from the exposure of Daphnia magna to the test item during the range-finding test are given in Table 1 (attached).
- No immobilisation was observed at 10 and 100 mg/L loading rate WAF, however immobilisation observed at 0.10 and 1.0 mg/L loading rate WAF.
- Sub lethal effects were observed at 24 hours in the 0.10, 1.0 and 10 mg/L loading rate WAFs.
- Sub lethal effects were trapping at the surface.
- Trapping at the surface was observed at 48 hours in the control, 0.10, 1.0 and 10 mg/L loading rate WAFs.
- Based on this information, a single loading rate WAF of four replicates (100 mg/L) was selected for the definitive test. This experimental design conforms to a 'limit test' to confirm that no immobilisation or adverse reactions to exposure were observed.
- Chemical analysis of the test preparations at 0 and 48 hours showed measured test concentrations of less than the limit of quantification (LOQ) of the analytical method which was determined to be 0.20 mg/L.
- Does not infer that no test item was in solution, just that any dissolved test item was at a concentration of less than the LOQ.

DEFINITIVE TEST - CHEMICAL ANALYSIS OF TEST LOADING RATES
- Chemical analysis of the fresh test preparations at 0 hours showed a measured test concentration of 1.72 mg/L.
- Chemical analysis of the aged test preparation at 48 hours showed a measured test concentration of 1.11 mg/L.
- The dissolved test item may have been one or several components of the test item. Given that toxicity cannot be attributed to a single component or mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

DEFINITIVE TEST IMMOBILISATION DATA
- Cumulative immobilisation data and other observations from exposure of Daphnia magna to the test item during the definitive test are given in Table 2 (attached).
- There was no immobilisation in 20 daphnids exposed to a 100 mg/L loading rate WAF for a period of 48 hours.
- One daphnid was observed to be immobilised in replicate 1 and was considered to be due to natural causes as less than 10% immobilization occurred.
- It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L.

DEFINITIVE TEST – Sub-Lethal Effects
-No sub-lethal effects of exposure were observed throughout the test.

DEFINITIVE TEST - VALIDATION CRITERIA
- The test was considered to be valid given that none of the control daphnids showed immobilisation or other signs of disease or stress and that the oxygen concentration at the end of the test was ≥ 3 mg/L in the control and test vessels.

DEFINITIVE TEST - WATER QUALITY CRITERIA
- The results of the water quality measurements are given in Table 3 (attached).
- Temperature was maintained at approximately 22 °C throughout the test.
- There were no treatment related differences for oxygen concentration or pH.
- Light intensity was observed to be in the range 602 to 633 lux throughout the test.

DEFINITIVE TEST - VORTEX DEPTH MEASUREMENTS
- The vortex depth recorded at the start and end of the mixing period was observed to be a dimple at the water surface on each occasion.

DEFINITIVE TEST - OBSERVATIONS ON TEST ITEM SOLUBILITY
- Observations were carried out during the mixing and testing of the WAF.
- At the start of the mixing period, the 100 mg/L loading rate was observed to be a clear colourless water column with visible lumps of yellow test item on the surface.
- After 23 hours stirring and a 1 hour standing period, the 100 mg/L loading rate was observed to remain a clear colourless water column with lumps of yellow test item on the surface.
- Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item present.
- After siphoning and during the test the 100 mg/L loading rate was observed to be a clear, colourless solution.

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item to the freshwater invertebrate Daphnia magna has been investigated and gave a 48-Hours EL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

Executive summary:

GUIDELINE

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202 "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No 440/2008.

METHODS

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF). Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to a Water Accommodated Fraction (WAF) of the test item at a nominal loading rate of 100 mg/L for 48 hours at a temperature of approximately 22°C under static test conditions. Immobilization and any adverse reactions to exposure were recorded after 24 and 48 hours.

RESULTS

Chemical analysis of the fresh test preparation at 0 hours showed a measured test concentration of 1.72 mg/L. Chemical analysis of the aged test preparation at 48 hours showed a measured test concentration of 1.11 mg/L. The dissolved test item may have been one or several components of the test item. Given that the toxicity cannot be attributed to a single component or mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

CONCLUSION

Exposure of Daphnia magna to the test item gave EL50 values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF. It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Reason / purpose for cross-reference:

read-across source

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Description of key information

Exposure of Daphnia magna to the analogue test item gave EL50 values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L (OECD 202 and EU Method C.2).

Additional information

GUIDELINE

A study was performed to assess the acute toxicity of an analogue test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202 "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No 440/2008.

METHODS

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to a Water Accommodated Fraction (WAF) of the test item at a nominal loading rate of 100 mg/L for 48 hours at a temperature of approximately 22°C under static test conditions. Immobilization and any adverse reactions to exposure were recorded after 24 and 48 hours.

RESULTS

Chemical analysis of the fresh test preparation at 0 hours showed a measured test concentration of 1.72 mg/L. Chemical analysis of the aged test preparation at 48 hours showed a measured test concentration of 1.11 mg/L.The dissolved test item may have been one or several components of the test item. Given that the toxicity cannot be attributed to a single component or mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

CONCLUSION

Exposure of Daphnia magna to the analogue test item gave EL50 values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF. It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.