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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
08.11.1995 to 10.11.1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
EU Regulation 92/69/EWG of 31.07.1992
Deviations:
not specified
Principles of method if other than guideline:
The study was conducted under special consideration of the VCI concept design for (acute) aquatic testing of poorly soluble substances of 31.1.1994.
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: HenkelKGaA, SIS no. 6239, RIS-No. 51Z1200071000
- Expiration date of the lot/batch: 10/1996
- Manufacturing date: 03/1995
- Analytical purity: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
Analytical monitoring:
yes
Details on sampling:
The analytical recovery of the test substance was confirmed at the beginning and end of the test.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the low solubility of the test substance, two different test solutions were prepared:
A) Direct addition of test item to test solution: the test substance was directly weighed into the vessel and ultrasonicated for 5 min
B) Water Accommodated Fraction (WAF): 100 mg of test substance was placed in 1L of M4 medium and stirred for 24 hours with a magnetic stirrer. The undissolved test substance was separated with a glass fibre filter (pre-treated with sodium hydroxide solution).

- Controls: M4 medium
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: waterflea
- Strain/clone: Daphnia magna Straus
- Age at study initiation (mean and range, SD): Daphnids 6 to 24-hour old were used in the test.
- Source: Testing facility, originally obtained from BGA Berliun (May 1994)
- Feeding during test : Daphnids were not fed during test.

ACCLIMATION
- Type and amount of food: Daphnids in the culture were fed with algae, Scenedesmus subspicatus, amount not reported.
- Feeding frequency: Not reported.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
Not reported.
Test temperature:
21.8°C
pH:
7.8 - 7.9
Dissolved oxygen:
Dissolved oxygen was ≥ 90% oxygen saturation.
Nominal and measured concentrations:
Nominal concentration: 100 mg/L
Measured concentrations in test solution A (directly weighed test item): 105.36 mg/L at 0 h, 44.30 mg/L at 48 h.
Measured concentrations in test solution B (WAF): 0.29 mg/L at 0 h, 0.15 mg/L at 48 h. The recovery of the test substance at the beginning and after 48 hours was higher than the water solubility of the test substance of < 0.01 mg/L (determined with the column elution method), which indicates that the separation of test item was not complete.
Details on test conditions:
TEST SYSTEM
- Test vessel: glass beaker
- Material, size, headspace, fill volume: 100 mL beakers covered with watch glasses
- Volume of solution: 50 mL
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: M4 medium according to Elendt

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light daily
- Light intensity: Not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation after 24 and 48 hours.
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: See 'Remark'
Remarks:
Based on immobilisation of daphnids in solution A (prepared as direct addition without separation of undissolved test substance)
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
WAF
Basis for effect:
mobility
Remarks on result:
other: See 'Remark'
Remarks:
Based on immobilisation of daphnids in solution B (prepared as Water Accomodated Fraction, with separation of undissolved test substance)
Details on results:
- Behavioural abnormalities: Not reported
- Mortality of control: No mortality in control observed.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: In solution A (prepared as direct addition without separation of undissolved test substance) black particles were observed, partly attached to the daphnids. A turbidity of the test suspension was observed.
- Effect concentrations exceeding solubility of substance in test medium: Solution A was prepared as direct addition without separation of undissolved test substance and was tested above the water solubility of test item.

Table 1. Immobilisation (%) of daphnids in solution A (prepared as direct addition without separation of undissolved test substance)

Concentration (mg/L) / replicate

Incubation time

24 h

48 h

0/1

0

0

0/2

0

0

100/1

0*

10*

100/2

10*

10*

*Black particles were observed in the suspension which were partially attached to the daphnids. A turbidity of the test suspension was observed.

 Table 2. Immobilisation (%) of daphnids in solution B (prepared as Water Accomodated Fraction, with separation of undissolved test substance)

Concentration (mg/L) / replicate

Incubation time

24 h

48 h

0/1

0

0

0/2

0

0

100/1

0

0

100/2

0

0

Validity criteria fulfilled:
yes
Conclusions:
The EL50 for Daphnia magna exposed to glyceryl trioleate was determined to be >100 mg/L loading rate WAF based on nominal test concentrations.
Executive summary:

The acute toxicity of glyceryl trioleate to Daphnia magna was investigated over 48 hours under static conditions, in a study conducted according to EU Method C.2 (Kuch 1995). The study was conducted as a limit test with one test concentration of 100 mg/L. The study gave consideration to the VCI concept for poorly soluble substances and therefore two different test solutions were prepared: one solution where the test substance was directly weighed into the test vessel and undissolved substance was not removed and a second solution with separation of undissolved substance through filtration (preparation of a Water Accommodated Fraction (WAF)). Test item concentrations were verified by chemical analysis using gas chromatography. Immobilisation of Daphnids was recorded after 24 and 48 hours of exposure.

After 48 hours no immobilisation of Daphnia was observed at 100 mg/L loading rate WAF and 10% immobilisation was observed in the oversaturated solution with undissolved test substance at 100 mg/L.

The study is a guideline study, is GLP-compliant and is considered to be reliable with restrictions (Klimisch 2).

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08.11.1995 to 10.11.1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
EU Regulation 92/69/EWG of 31.07.1992
Deviations:
not specified
Principles of method if other than guideline:
The study was conducted under special consideration of the VCI concept design for (acute) aquatic testing of poorly soluble substances of 31.1.1994.
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: HenkelKGaA, SIS no. 6239, RIS-No. 51Z1200071000
- Expiration date of the lot/batch: 10/1996
- Manufacturing date: 03/1995
- Analytical purity: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
Analytical monitoring:
yes
Details on sampling:
The analytical recovery of the test substance was confirmed at the beginning and end of the test.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the low solubility of the test substance, two different test solutions were prepared:
A) Direct addition of test item to test solution: the test substance was directly weighed into the vessel and ultrasonicated for 5 min
B) Water Accommodated Fraction (WAF): 100 mg of test substance was placed in 1L of M4 medium and stirred for 24 hours with a magnetic stirrer. The undissolved test substance was separated with a glass fibre filter (pre-treated with sodium hydroxide solution).

- Controls: M4 medium
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: waterflea
- Strain/clone: Daphnia magna Straus
- Age at study initiation (mean and range, SD): Daphnids 6 to 24-hour old were used in the test.
- Source: Testing facility, originally obtained from BGA Berliun (May 1994)
- Feeding during test : Daphnids were not fed during test.

ACCLIMATION
- Type and amount of food: Daphnids in the culture were fed with algae, Scenedesmus subspicatus, amount not reported.
- Feeding frequency: Not reported.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
Not reported.
Test temperature:
21.8°C
pH:
7.8 - 7.9
Dissolved oxygen:
Dissolved oxygen was ≥ 90% oxygen saturation.
Nominal and measured concentrations:
Nominal concentration: 100 mg/L
Measured concentrations in test solution A (directly weighed test item): 105.36 mg/L at 0 h, 44.30 mg/L at 48 h.
Measured concentrations in test solution B (WAF): 0.29 mg/L at 0 h, 0.15 mg/L at 48 h. The recovery of the test substance at the beginning and after 48 hours was higher than the water solubility of the test substance of < 0.01 mg/L (determined with the column elution method), which indicates that the separation of test item was not complete.
Details on test conditions:
TEST SYSTEM
- Test vessel: glass beaker
- Material, size, headspace, fill volume: 100 mL beakers covered with watch glasses
- Volume of solution: 50 mL
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: M4 medium according to Elendt

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light daily
- Light intensity: Not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation after 24 and 48 hours.
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: See 'Remark'
Remarks:
Based on immobilisation of daphnids in solution A (prepared as direct addition without separation of undissolved test substance)
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
WAF
Basis for effect:
mobility
Remarks on result:
other: See 'Remark'
Remarks:
Based on immobilisation of daphnids in solution B (prepared as Water Accomodated Fraction, with separation of undissolved test substance)
Details on results:
- Behavioural abnormalities: Not reported
- Mortality of control: No mortality in control observed.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: In solution A (prepared as direct addition without separation of undissolved test substance) black particles were observed, partly attached to the daphnids. A turbidity of the test suspension was observed.
- Effect concentrations exceeding solubility of substance in test medium: Solution A was prepared as direct addition without separation of undissolved test substance and was tested above the water solubility of test item.

Table 1. Immobilisation (%) of daphnids in solution A (prepared as direct addition without separation of undissolved test substance)

Concentration (mg/L) / replicate

Incubation time

24 h

48 h

0/1

0

0

0/2

0

0

100/1

0*

10*

100/2

10*

10*

*Black particles were observed in the suspension which were partially attached to the daphnids. A turbidity of the test suspension was observed.

 Table 2. Immobilisation (%) of daphnids in solution B (prepared as Water Accomodated Fraction, with separation of undissolved test substance)

Concentration (mg/L) / replicate

Incubation time

24 h

48 h

0/1

0

0

0/2

0

0

100/1

0

0

100/2

0

0

Validity criteria fulfilled:
yes
Conclusions:
The EL50 for Daphnia magna exposed to glyceryl trioleate was determined to be >100 mg/L loading rate WAF based on nominal test concentrations.
Executive summary:

The acute toxicity of glyceryl trioleate to Daphnia magna was investigated over 48 hours under static conditions, in a study conducted according to EU Method C.2 (Kuch 1995). The study was conducted as a limit test with one test concentration of 100 mg/L. The study gave consideration to the VCI concept for poorly soluble substances and therefore two different test solutions were prepared: one solution where the test substance was directly weighed into the test vessel and undissolved substance was not removed and a second solution with separation of undissolved substance through filtration (preparation of a Water Accommodated Fraction (WAF)). Test item concentrations were verified by chemical analysis using gas chromatography. Immobilisation of Daphnids was recorded after 24 and 48 hours of exposure.

After 48 hours no immobilisation of Daphnia was observed at 100 mg/L loading rate WAF and 10% immobilisation was observed in the oversaturated solution with undissolved test substance at 100 mg/L.

The study is a guideline study, is GLP-compliant and is considered to be reliable with restrictions (Klimisch 2).

Description of key information

The EL50 for Daphnia magna is >100 mg/L loading rate WAF based on nominal test concentrations.

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
100 mg/L

Additional information

No short-term data on toxicity of Tallicin K2 to aquatic invertebrates is available. Therefore, a study with glyceryl trioleate is used to complete this endpoint. The study was conducted according to a standard guideline, is GLP-compliant and is considered to be reliable with restrictions (Klimisch 2).

Acute toxicity of glyceryl trioleate to Daphnia magna was investigated in a limit test conducted according to EU method C.2. The study was conducted with additional consideration of the VCI concept for poorly soluble substances and therefore two different test solutions were prepared: one solution where the test substance was directly weighed into the test vessel and undissolved substance was not removed and a second solution with separation of undissolved substance through filtration (preparation of a Water Accommodated Fraction (WAF)).

After 48 hours no immobilisation of Daphnia was observed at 100 mg/L loading rate WAF and 10% immobilisation was observed in the oversaturated solution with undissolved test substance at 100 mg/L.