Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
05/11/2012 - 04/01/2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study according to international guideline (OECD guideline 202) under GLP.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not relevant
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 100 mg/L
- Sampling method: Water samples were taken from the control and the 100 mg/L loading rate WAF test group (replicates R1 - R4 pooled) at 0 and 48 hours
- Sample storage conditions before analysis: Samples were stored at approximately -20 °C prior to analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: An amount of test item (200 mg) was added to the surface of 2 liters of reconstituted water to give the 100 mg/L loading rate. After the addition of the test item, the reconstituted water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixture allowed to stand for 1 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 inspection of the WAF showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 100 mg/L loading rate WAF.
- Controls: yes, blanks
- Observations on solubility: At the start of the mixing period the 100 mg/L loading rate was observed to be a clear colorless water column with oily globules of 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 colorless water column with oily globules of test item on the surface. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 100 mg/L loading rate was observed to be a clear, colorless solution.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Source: in-house laboratory culture
- Method of breeding: Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium (see Appendix 1) in a temperature controlled room at approximately 20 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition 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, such that the 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 are considered not to contain any contaminant that would affect the integrity or outcome of the study.
- Feeding during test: no
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
Not relevant
Hardness:
250 mg/L as CaCO3
Test temperature:
20 - 21ºC
pH:
7.9 - 8.1
Dissolved oxygen:
8.8 - 9.3 mg/L ( 97 - 102% of ASV)
Salinity:
Not relevant
Nominal and measured concentrations:
Nominal: 100 mg/l
Measured (TOC): 1.21 mg C/L (0 h) and < LoQ (48 h)
Details on test conditions:
TEST SYSTEM
- Test vessel: jar
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: glass, 250 mL filled with 200 mL test medium
- Aeration: None
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted wtaer
- Culture medium different from test medium: No, Elendt M7 medium
- Intervals of water quality measurement: pH, temperature and oxygen concentration were measured every 24 h, starting at 0 h

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Test was performed in the dark, to avoid exposure of the test substance to light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mobility (every 24h)

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Duration:
48 h
Dose descriptor:
NOELR
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:
24 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Mortality of control: 0%
- Abnormal responses:
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
- Effect concentrations exceeding solubility of substance in test medium: At the start of the mixing period the 100 mg/L loading rate was observed to be a clear colorless water column with oily globules of 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 colorless water column with oily globules of test item on the surface. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 100 mg/L loading rate was observed to be a clear, colorless solution.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- EC50/LC50: 48h-EC50: 0.45 mg/L
Reported statistics and error estimates:
An estimate of the EL*50 values was given by inspection of the immobilization data.

For detailed results, see attached file "Results.docx".

Validity criteria fulfilled:
yes
Remarks:
Mortality in controls <10%, physico-chemical parameters within acceptable limits
Conclusions:
The acute toxicity (48h-EL50) of soybean oil, polymerized measured in a WAF test is > 100 mg/l.
Executive summary:

The acute toxicity of soybean oil, polymerized was investigated according to OECD guideline 202 under GLP. Daphnids were exposed to a WAF of 100 mg/l and observed for 48 hours. The 48h-NOEL and 48h-EL50 were found to be 100 and > 100 mg/l respectively.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-11-28 - 2013-12-15
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Read-across to effluent test, no GLP, in line with OECD 202 but no Standard guidance.
Reason / purpose:
reference to other study
Reason / purpose:
reference to same study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
Test material was the effluent of a SCAS unit fed with domestic wastewater and Stand Linseed Oil. Thus the toxicity of the biodegradation products of Stand Linseed Oil was tested.
GLP compliance:
no
Remarks:
The essence of the test was to investigate the toxicity of the effluent of the SCAS test to daphnids in an experimental setup. No GLP protocol based on existing SOPs for such set-up was available.
Analytical monitoring:
no
Details on test solutions:
Dilutions of the SCAS effluent containing biodegradation product(s) of Linseed standoil were made with effluent from the control SCAS unit.
Control groups included:
1. DSW (Dutch Standard Water) : check for normal performance of the Daphnia magna
2. Effluent of Control SCAS Unit : used as dilution medium; check for absence of effects on the test organisms
3. Effluent Control SCAS Unit with 50 mg Tween 80/L: check for effects of the dispersant, if still present
5. Effluent Control SCAS Unit with 20 mg Stand Linseed Oil/L and 20 mg Tween 80/L : check for effects of non-degraded parent materials
6. Effluent Control SCAS Unit with 50 mg Stand Linseed Oil/L and 50 mg Tween 80/L : check for effects of non-degraded parent materials

The medium containing the actual test material was:
7. Effluent of Linseed Standoil SCAS Unit

The test medium Dutch Standard Water contains per litre demineralised water 200 mg of CaCl2.2H2O, 180 mg of MgSO4.7H2O, 100 mg NaHCO3 and 20 mg KHCO3. The pH of DSW ranges from 7.0 – 8.0.

Test organisms (species):
Daphnia magna
Details on test organisms:
Aged less than 24 h at the start of the test, taken from a stock culture of 2 to 4 weeks old
Test type:
static
Water media type:
freshwater
Total exposure duration:
48 h
Test temperature:
20 - 21 ºC
pH:
6.7 - 7.2 at start of test
6.7 - 7.4 after 48 h
Dissolved oxygen:
Fullfilled the requirement that oxygen concentration should be at or above 3 mg/l in control and test vessels
Nominal and measured concentrations:
The collected effluent of the SCAS unit fed with Linseed standoil (mixed effluent sample) contained 15 mg/L organic carbon most likely derived from Linseed standoil biodegradation product(s). The most likely molecular formula of the degradation product of Linseed standoil is C36O4H64 (see IUCLID entry 5.2.1). Using this formula a carbon content of 0.77 g carbon/g degradation product can be calculated and subsequently a concentration of 19.5 mg degradation product/L effluent. The lowest dilution factor (20 times) gives a degradation product concentration of approximately 1 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 ml-glass beakers
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 50 ml of medium
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 1 replicate in preliminary test, 2 replicates (10 animals) in final test
- No. of vessels per control (replicates): 2 replicates
- No. of vessels per vehicle control (replicates): 1 or 2 replicates each for each of the controls

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dutch Standard Water DSW or effluent of the SCAS units
- Total organic carbon: 18 - 26 mg/L (Non purgeable Organic Carbon)

TEST CONCENTRATIONS
- Test concentrations: Based on measured Organic Carbon content and converted to mass of biodegradation products.
Preliminary test: undiluted effluent: circa 19.5 mg degradation products/L;
Final test: effluent dilutions with factor 3.2, so 1:20 - 1: 64 - 1:200 - 1: 640 - 1: 2000 and undiluted effluent
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 1 mg/L
Nominal / measured:
estimated
Conc. based on:
other: total mass of biodegradation products
Basis for effect:
mobility
Details on results:
- Mortality of control: complete survival of all daphnids in all controls

Test Results

Preliminary study: number of mobile daphnids in different test media

Medium

Number of mobile daphnids

0 hours

24 hours

48 hours

Control: Effluent SCAS unit

 

5

5

5

Effluent Linseed standoil SCAS unit

 

5

3

0

Control: DSW

 

5

5

5

Control: Effluent SCAS unit supplemented with 20 mg/L Linseed standoil and 20 mg/L Tween 80 *

5

5

5

Control: Effluent SCAS unit supplemented with 50 mg/L Linseed standoil and 50 mg/L Tween 80*

5

5

5

Control: Effluent SCAS unit supplemented with 50 mg/L Tween 80

5

5

5

Effluent supplemented with Linseed Stand Oil and Tween 80 were light suspensions


Definitive study: number of mobile daphnids in different test media

Medium

Replicates

Number of mobile daphnids

0 hours

24 hours

48 hours

Control: Effluent SCAS unit

 

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit

 

I

II

5

5

3

3

0

0

Control: DSW

 

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:20 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:64 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:200 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:640 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:2000 dilution)

I

II

5

5

5

5

5

5

Dilutions of effluent from the Linseed standoil SCAS unit were made with effluent from the control SCAS unit.

Validity criteria fulfilled:
yes
Remarks:
Validation criteria were met: no effects in the control and oxygen concentrations remained > 3 mg/l in all test concentrations
Conclusions:
The toxicity of the biodegradation products of Stand Linseed Oil, expressed as the 48h-EC50, is > 1 mg (total mass) per Liter.
Executive summary:

The biodegradation of Stand Linseed Oil was studied in a Semi-Continuous Activated Sludge (SCAS) system (OECD TG 302). The test substance did not biodegrade completely, so the effluent of the SCAS system contained biotransformation products. Based on the measured Organic Carbon content of the effluent, the total concentration of the biotransformation products was estimated at 19.5 mg/l. The parent substance is not toxic to Daphnia, but the undiluted effluent caused complete mortality within 2 days. Yet in a 1:20 dilution of the effluent no effects were observed after 48 h. This implies that the 48h-EC50 of the (total mass of the) biodegradation products is above 1 mg/L.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
See attached justification
Reason / purpose:
reference to other study
Reason / purpose:
read-across source
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 1 mg/L
Nominal / measured:
estimated
Conc. based on:
other: total mass of biodegradation products
Basis for effect:
mobility
Details on results:
- Mortality of control: complete survival of all daphnids in all controls

Test Results

Preliminary study: number of mobile daphnids in different test media

Medium

Number of mobile daphnids

0 hours

24 hours

48 hours

Control: Effluent SCAS unit

 

5

5

5

Effluent Linseed standoil SCAS unit

 

5

3

0

Control: DSW

 

5

5

5

Control: Effluent SCAS unit supplemented with 20 mg/L Linseed standoil and 20 mg/L Tween 80 *

5

5

5

Control: Effluent SCAS unit supplemented with 50 mg/L Linseed standoil and 50 mg/L Tween 80*

5

5

5

Control: Effluent SCAS unit supplemented with 50 mg/L Tween 80

5

5

5

Effluent supplemented with Linseed Stand Oil and Tween 80 were light suspensions


Definitive study: number of mobile daphnids in different test media

Medium

Replicates

Number of mobile daphnids

0 hours

24 hours

48 hours

Control: Effluent SCAS unit

 

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit

 

I

II

5

5

3

3

0

0

Control: DSW

 

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:20 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:64 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:200 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:640 dilution)

I

II

5

5

5

5

5

5

Effluent Linseed standoil SCAS unit (1:2000 dilution)

I

II

5

5

5

5

5

5

Dilutions of effluent from the Linseed standoil SCAS unit were made with effluent from the control SCAS unit.
Validity criteria fulfilled:
yes
Remarks:
Validation criteria were met: no effects in the control and oxygen concentrations remained > 3 mg/l in all test concentrations
Conclusions:
The toxicity of the biodegradation products of Stand Linseed Oil, expressed as the 48h-EC50, is > 1 mg (total mass) per Liter.
Executive summary:

The biodegradation of Stand Linseed Oil was studied in a Semi-Continuous Activated Sludge (SCAS) system (OECD TG 302). The test substance did not biodegrade completely, so the effluent of the SCAS system contained biotransformation products. Based on the measured Organic Carbon content of the effluent, the total concentration of the biotransformation products was estimated at 19.5 mg/l. The parent substance is not toxic to Daphnia, but the undiluted effluent caused complete mortality within 2 days. Yet in a 1:20 dilution of the effluent no effects were observed after 48 h. This implies that the 48h-EC50 of the (total mass of the) biodegradation products is above 1 mg/L.

Description of key information

48h-EL50: > 100 mg/L

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
100 mg/L

Additional information

The acute toxicity of soybean oil, polymerized was investigated according to OECD guideline 202 under GLP. Daphnids were exposed to a WAF of 100 mg/l and observed for 48 hours. The 48h-NOEL and 48h-EL50 were found to be 100 and > 100 mg/l respectively.

A study on the acute toxicity to Daphnia magna (according to OECD 202, not under GLP) was also conducted with the biodegradation products of Linseed oil, polymerized. This result is read-across to the biodegradation products of Soybean oil, polymerized. Based on the results of this test, it was concluded that the 48h-EC50 of the (total mass of the) biodegradation products is above 1 mg/L. Based on this result it can be concluded that the biodegradation products of Soybean oil, polymerized do not meet the "T" criterion in the PBT assessment.