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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Study period:
2014-05-23 to 2014-05-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: QSAR value. The substance falls into the applicability domains of the QSAR model.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
QSAR model
Principles of method if other than guideline:
The acute toxicity to aquatic invertebrates was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
no
Details on test solutions:
not applicable
Test organisms (species):
Daphnia sp.
Details on test organisms:
not applicable
Test type:
other: QSAR
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
48h-EC50
Post exposure observation period:
not applicable
Hardness:
The QSAR is based on data from studies performed at acceptable hardness to ensure control survival
Test temperature:
The QSAR is based on data from studies performed at between 14 - 25 °C (depending on the species considered)
pH:
The QSAR is based on data from studies performed at acceptable pH between 6.0 - 9.0
Dissolved oxygen:
The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%)
Salinity:
The QSAR is based on data from studies performed on freshwater species
Nominal and measured concentrations:
The QSAR is based on data from studies performed using measured concentrations or with acceptable stability
Details on test conditions:
not applicable
Reference substance (positive control):
no
Remarks:
QSAR model
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.65 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL [0.40-1.06]
Details on results:
not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
95% CL [0.40-1.06]
QSAR statistical parameters are given in the QMRF and the QPRF

no data

Validity criteria fulfilled:
yes
Remarks:
The substance falls into the applicability domains of the QSAR model.
Conclusions:
48h-EC50 for myrcene = 0.65 mg test item/L with 95% Confidence Limit between 0.40 and 1.06 mg test item/L.
Executive summary:

The acute toxicity to daphnia was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.

The acute toxicity to daphnia of myrcene has been investigated using a QSAR model that predicts daphnia mortality in an OECD 202 study. Myrcene falls within the applicability domain of the model as demonstrated in the QPRF.

The 48-h EC50 was 0.65 mg test material/L with 95%-Confidence Limit between 0.40 and 1.06 mg test material/L. EC50 were based on mobility. 

Based on the results of this study, myrcene would be classified as acute 1 to aquatic organisms in accordance with the classification of the CLP.

This toxicity study is acceptable and can be used for that endpoint.

 

Results Synopsis

Test Type: QSAR model

LC50: 0.65 mg test material/L 95%-C.L.: 0.40 to 1.06 mg test material/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Study period:
2014-05-23 to 2014-05-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: QSAR value. The substance falls into the applicability domains of the QSAR model.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
QSAR model
Principles of method if other than guideline:
The acute toxicity to aquatic invertebrates was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
no
Details on test solutions:
not applicable
Test organisms (species):
Daphnia sp.
Details on test organisms:
not applicable
Test type:
other: QSAR
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
48h-EC50
Post exposure observation period:
not applicable
Hardness:
The QSAR is based on data from studies performed at acceptable hardness to ensure control survival
Test temperature:
The QSAR is based on data from studies performed at between 14 - 25 °C (depending on the species considered)
pH:
The QSAR is based on data from studies performed at acceptable pH between 6.0 - 9.0
Dissolved oxygen:
The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%)
Salinity:
The QSAR is based on data from studies performed on freshwater species
Nominal and measured concentrations:
The QSAR is based on data from studies performed using measured concentrations or with acceptable stability
Details on test conditions:
not applicable
Reference substance (positive control):
no
Remarks:
QSAR model
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.57 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL [0.34-0.94]
Details on results:
not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
95% CL [0.34-0.94]
QSAR statistical parameters are given in the QMRF and the QPRF

no data

Validity criteria fulfilled:
yes
Remarks:
The substance falls into the applicability domains of the QSAR model.
Conclusions:
48h-EC50 for Δ-3-carene = 0.57 mg test item/L with 95% Confidence Limit between 0.34 and 0.94 mg test item/L.
Executive summary:

The acute toxicity to daphnia was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.

The acute toxicity to daphnia of Δ-3-carene has been investigated using a QSAR model that predicts daphnia mortality in an OECD 202 study. Δ-3-carene falls within the applicability domain of the model as demonstrated in the QPRF.

The 48-h EC50 was 0.57 mg test material/L with 95%-Confidence Limit between 0.34 and 0.94 mg test material/L. EC50 were based on mobility. 

Based on the results of this study, Δ-3-carene would be classified as acute 1 to aquatic organisms in accordance with the classification of the CLP.

This toxicity study is acceptable and can be used for that endpoint.

 

Results Synopsis

Test Type: QSAR model

LC50: 0.57 mg test material/L 95%-C.L.: 0.34 to 0.94 mg test material/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Study period:
2014-05-20 to 2014-05-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: QSAR value. The substance falls into the applicability domains of the QSAR model.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
QSAR model
Principles of method if other than guideline:
The acute toxicity to aquatic invertebrates was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
no
Details on test solutions:
not applicable
Test organisms (species):
Daphnia sp.
Details on test organisms:
not applicable
Test type:
other: QSAR
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
48h-EC50
Post exposure observation period:
not applicable
Hardness:
The QSAR is based on data from studies performed at acceptable hardness to ensure control survival
Test temperature:
The QSAR is based on data from studies performed at between 14 - 25 °C (depending on the species considered)
pH:
The QSAR is based on data from studies performed at acceptable pH between 6.0 - 9.0
Dissolved oxygen:
The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%)
Salinity:
The QSAR is based on data from studies performed on freshwater species
Nominal and measured concentrations:
The QSAR is based on data from studies performed using measured concentrations or with acceptable stability
Details on test conditions:
not applicable
Reference substance (positive control):
no
Remarks:
QSAR model
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
1.37 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL [0.91-2.07]
Details on results:
not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
95% CL [0.91-2.07]
QSAR statistical parameters are given in the QMRF and the QPRF

no data

Validity criteria fulfilled:
yes
Remarks:
The substance falls into the applicability domains of the QSAR model.
Conclusions:
48h-EC50 for limonene = 1.37 mg test item/L with 95% Confidence Limit between 0.91 and 2.07 mg test item/L.
Executive summary:

The acute toxicity to daphnia was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.

The acute toxicity to daphnia of limonene has been investigated using a QSAR model that predicts daphnia mortality in an OECD 202 study. Limonene falls within the applicability domain of the model as demonstrated in the QPRF.

The 48-h EC50 was 1.37 mg test material/L with 95%-Confidence Limit between 0.91 and 2.07 mg test material/L. EC50 were based on mobility. 

Based on the results of this study, limonene would not be classified as acute 1 to aquatic organisms in accordance with the classification of the CLP.

This toxicity study is acceptable and can be used for that endpoint.

 

Results Synopsis

Test Type: QSAR model

LC50: 1.37 mg test material/L 95%-C.L.: 0.91 to 2.07 mg test material/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted similarly to OECD guideline 202 with minor deviations: no data on acclimation period, feeding pattern and reference substance. There are no details on the solubility data of the substance and this value is not in accordance with the solubility determined in a recent study conducted according to the OECD guideline. In fact, results show that there is no toxicity at the solubilit limit. Mortality at higher nominal rates could be explain by physical effects.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
no data on acclimation period, feeding pattern and reference substance
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No data
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0, 390, 780, 1170, 1560 or 1950 µg/L
- Sampling time: 0, 24 and 48 hours
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test solutions was provided as liquid/liquid saturation units by adding excess of test chemical in 14 L of Lake Superior water
- Controls: Dilution water
- Chemical name of vehicle: Water
- Concentration of vehicle in test medium: Stock (1950 µg/L liq-liq equil.)
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Source: ERL-Duluth, USA
- Age at study initiation: < 24 hours
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
None
Hardness:
43.8 ± 1.2 mg/L CaCO3
Test temperature:
20.1 ± 0.4 °C
pH:
7.9 ± 0.05
Dissolved oxygen:
8.2 ± 0.2 mg O2/L
Salinity:
No data
Nominal and measured concentrations:
Nominal concentrations: 0, 390, 780, 1170, 1560 and 1950 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Six crystallizing dishes were used as test chambers in the electronic diluter with tank volumes of 250 mL; test solution depth: 4-5 cm
- Type (delete if not applicable): Closed; stainless steel screen enclosures (forty mesh) were placed in each chamber for the daphnia exposures
- Type of flow-through: Electronic diluter
- Renewal rate of test solution (frequency/flow rate): Flow rates of 21 mL/min to each tank provided concentrations of 244-1760 µg/L
- No. of organisms per vessel: 15
- No. of vessels per concentration (replicates): One
- No. of vessels per vehicle control (replicates): One

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Lake Superior water
- Preparation of dilution water: Water was passed through sand filters, ultraviolet sterilizers and 55 µm fibre filter. Test waters were then aerated and heated at 20 ± 1 °C
- Alkalinity: 36.2 ± 0.3 mg/L CaCO3
- Intervals of water quality measurement: Tank temperatures (°C), dissolved oxygen (mg/L) and pH were measured daily in each of the test chambers where test organisms were still alive

OTHER TEST CONDITIONS
- Photoperiod: 16 hour/day light cycle; cool white fluorescent light
- Light intensity: 12-28 lumens

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Mortalities or other effects were observed at 24 and 48 hours
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
1 250 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 1150-1360 µg/L
Details on results:
- Mortality or behavioural abnormalities: No mortality occurred during the test, however, organisms showed signs of stress at approximately 24 hour, failed to swim throughout the water column and remained on the bottoms of the test chambers. Because the highest chamber concentration was measured near water solubility (approximately 2100 µg/L), the test was not repeated.
- LC50 values were not determined due to limited mortality
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Measured concentrations were lower than nominal concentrations due to volatility.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
- EC50 was calculated using the corrected average of the analyzed tank concentrations and the Trimmed Spearman-Karber Method (Hamilton et al., 1977).
- Mean toxicant concentrations used in the calculations were corrected for analytical spike recoveries

Table 2: Effects

 

Hour

Control (0 µg/L)

A (390 µg/L)

B (780 µg/L)

C (1170 µg/L)

D (1560 µg/L)

E (1950 µg/L)

Initial

15

15

15

15

15

15

24

 

 

 

 

 

15

48

 

 

 

 

5

15

Validity criteria fulfilled:
yes
Remarks:
mortality in control < 10% (no immobilisation observed); dissolved oxygen concentration > 3 mg O2/L
Conclusions:
Under the test conditions, no effects were observed at the solubility limit.
Executive summary:

In an acute aquatic toxicity study performed similarly to OECD guideline 202, groups (15/dose) of Daphnia magna was exposed to β-pinene at concentrations of 0, 390, 780, 1170, 1560 and 1950 µg/L (nominal) for 48 hours under flow-through conditions. Mortality and effects were observed at 24 and 48 hours.

 

No mortality occurred during the test, however, organisms showed signs of stress at approximately 24 hour, failed to swim throughout the water column and remained on the bottoms of the test chambers.

The study was conducted similarly to OECD guideline 202 but there are no details on the solubility data of the substance and this value is not in accordance with the solubility determined in a recent study conducted according to the OECD guideline. In fact, results show that there is no toxicity at the solubilit limit. Mortality at higher nominal rates could be explain by physical effects.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Study period:
2014-05-23 to 2014-05-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: QSAR value. The substance falls into the applicability domains of the QSAR model.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
QSAR model
Principles of method if other than guideline:
The acute toxicity to aquatic invertebrates was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
no
Details on test solutions:
not applicable
Test organisms (species):
Daphnia sp.
Details on test organisms:
not applicable
Test type:
other: QSAR
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
48h-EC50
Post exposure observation period:
not applicable
Hardness:
The QSAR is based on data from studies performed at acceptable hardness to ensure control survival
Test temperature:
The QSAR is based on data from studies performed at between 14 - 25 °C (depending on the species considered)
pH:
The QSAR is based on data from studies performed at acceptable pH between 6.0 - 9.0
Dissolved oxygen:
The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%)
Salinity:
The QSAR is based on data from studies performed on freshwater species
Nominal and measured concentrations:
The QSAR is based on data from studies performed using measured concentrations or with acceptable stability
Details on test conditions:
not applicable
Reference substance (positive control):
no
Remarks:
QSAR model
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.8 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL [0.50-1.28]
Details on results:
not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
95% CL [0.50-1.28]
QSAR statistical parameters are given in the QMRF and the QPRF

no data

Validity criteria fulfilled:
yes
Remarks:
The substance falls into the applicability domains of the QSAR model.
Conclusions:
48h-EC50 for β-phellandrene = 0.80 mg test item/L with 95% Confidence Limit between 0.50 and 1.28 mg test item/L.
Executive summary:

The acute toxicity to daphnia was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.

The acute toxicity to daphnia of β-phellandrene has been investigated using a QSAR model that predicts daphnia mortality in an OECD 202 study. β-phellandrene falls within the applicability domain of the model as demonstrated in the QPRF.

The 48-h EC50 was 0.80 mg test material/L with 95%-Confidence Limit between 0.50 and 1.28 mg test material/L. EC50 were based on mobility. 

Based on the results of this study, Camphene would be classified as acute 1 to aquatic organisms in accordance with the classification of the CLP.

This toxicity study is acceptable and can be used for that endpoint.

 

Results Synopsis

Test Type: QSAR model

LC50: 0.80 mg test material/L 95%-C.L.: 0.50 to 1.28 mg test material/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Study period:
2014-05-23 to 2014-05-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: QSAR value. The substance falls into the applicability domains of the QSAR model.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
QSAR model
Principles of method if other than guideline:
The acute toxicity to aquatic invertebrates was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
no
Details on test solutions:
not applicable
Test organisms (species):
Daphnia sp.
Details on test organisms:
not applicable
Test type:
other: QSAR
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
48h-EC50
Post exposure observation period:
not applicable
Hardness:
The QSAR is based on data from studies performed at acceptable hardness to ensure control survival
Test temperature:
The QSAR is based on data from studies performed at between 14 - 25 °C (depending on the species considered)
pH:
The QSAR is based on data from studies performed at acceptable pH between 6.0 - 9.0
Dissolved oxygen:
The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%)
Salinity:
The QSAR is based on data from studies performed on freshwater species
Nominal and measured concentrations:
The QSAR is based on data from studies performed using measured concentrations or with acceptable stability
Details on test conditions:
not applicable
Reference substance (positive control):
no
Remarks:
QSAR model
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
7.28 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL [4.56-11.64]
Details on results:
not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
95% CL [4.56-11.64]
QSAR statistical parameters are given in the QMRF and the QPRF

no data

Validity criteria fulfilled:
yes
Remarks:
The substance falls into the applicability domains of the QSAR model.
Conclusions:
48h-EC50 for Bornyl acetate = 7.28 mg test item/L with 95% Confidence Limit between 4.56 and 11.64 mg test item/L.
Executive summary:

The acute toxicity to daphnia was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.

The acute toxicity to daphnia of Bornyl acetate has been investigated using a QSAR model that predicts daphnia mortality in an OECD 202 study. Bornyl acetate falls within the applicability domain of the model as demonstrated in the QPRF.

The 48-h EC50 was 7.28 mg test material/L with 95%-Confidence Limit between 4.56 and 11.64 mg test material/L. EC50 were based on mobility. 

Based on the results of this study, limonene would not be classified as acute 1 to aquatic organisms in accordance with the classification of the CLP.

This toxicity study is acceptable and can be used for that endpoint.

 

Results Synopsis

Test Type: QSAR model

LC50: 7.28 mg test material/L 95%-C.L.: 4.56 to 11.64 mg test material/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted similarly to OECD guideline 202 with minor deviations: no data on acclimation period, feeding pattern and reference substance.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
no data on acclimation period, feeding pattern and reference substance
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0, 124, 248, 372, 496 or 620 µg/L
- Sampling time: 0, 24 and 48 hours
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test solutions was provided as liquid/liquid saturation units by adding excess of test chemical in 14 L of Lake Superior water
- Controls: Dilution water
- Chemical name of vehicle: Water
- Concentration of vehicle in test medium: Stock (620 µg/L liq-liq equil.)
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Source: ERL-Duluth, USA
- Age at study initiation: < 24 hours
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
None
Hardness:
45.4 ± 0.2 mg/L CaCO3
Test temperature:
20.2 ± 0.2 °C
pH:
7.8 ± 0.09
Dissolved oxygen:
7.8 ± 0.4 mg O2/L
Salinity:
No data
Nominal and measured concentrations:
Nominal concentrations: 0, 124, 248, 372, 496 and 620 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Six crystallizing dishes were used as test chambers in the electronic diluter with tank volumes of 200 mL; test solution depth: 4-5 cm
- Type (delete if not applicable): Closed; stainless steel screen enclosures (forty mesh) were placed in each chamber for the daphnia exposures
- Type of flow-through: Electronic diluter
- Renewal rate of test solution (frequency/flow rate): Single tank exposures were conducted at reduced flow rates (< 7 mL/min) so that a higher concentration could be attained; two additional toxicity tests were completed using the single-cell toxicity screening system: first test utilized a 10 mL/min flow rate into the chamber, while the second test was set at 1 mL/min.
- No. of organisms per vessel: 15
- No. of vessels per concentration (replicates): One
- No. of vessels per vehicle control (replicates): One

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Lake Superior water
- Preparation of dilution water: Water was passed through sand filters, ultraviolet sterilizers and 55 µm fibre filter. Test waters were then aerated and heated at 20 ± 1 °C
- Alkalinity: 42.9 ± 0.1 mg/L CaCO3
- Intervals of water quality measurement: Tank temperatures (°C), dissolved oxygen (mg/L) and pH were measured daily in each of the test chambers where test organisms were still alive

OTHER TEST CONDITIONS
- Photoperiod: 16 hours/day light cycle; cool white fluorescent light
- Light intensity: 12-28 lumens

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Mortalities or other effects were observed at 24 and 48 hours
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
ca. 1 440 µg/L
Nominal / measured:
not specified
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- An approximate LC50 value of 1440 µg/L was calculated based on one mortality in the first test and total mortality in the second test
- EC50 values were not obtained
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
- LC50/EC50 was calculated using the corrected average of the analyzed tank concentrations and the Trimmed Spearman-Karber Method (Hamilton et al., 1977).
- Mean toxicant concentrations used in the calculations were corrected for analytical spike recoveries

Table 2: Mortalities

 

Hour

Control (0 µg/L)

A (124 µg/L)

B (248 µg/L)

C (372 µg/L)

D (496 µg/L)

E (620 µg/L)

Initial

15

15

15

15

15

15

48 (Deads)

 0

2

 

 

Table 2: Effects

 

Hour

Control (0 µg/L)

A (124 µg/L)

B (248 µg/L)

C (372 µg/L)

D (496 µg/L)

E (620 µg/L)

Initial

15

15

15

15

15

15

48 (Effects)

 0

 0

2

Validity criteria fulfilled:
yes
Remarks:
mortality in control < 10% (no immobilisation observed); dissolved oxygen concentration > 3 mg O2/L
Conclusions:
Under the test conditions, an approximate LC50 value of 1440 µg/L was calculated based on one mortality in the first test and total mortality in the second test.
Executive summary:

In an acute aquatic toxicity study performed similarly to OECD guideline 202, groups (15/dose) of Daphnia magna were exposed to (+)-alpha pinene at concentrations of 0, 124, 248, 372, 496 and 620 µg/L (nominal) for 48 hours under flow-through conditions. Mortality and effects were observed at 24 and 48 hours.

An approximate EC50 value of 1440 µg/L was calculated based on one mortality in the first test and total mortality in the second test. EC50 was not obtained.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Study period:
2014-05-23 to 2014-05-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: QSAR value. The substance falls into the applicability domains of the QSAR model.
Justification for type of information:
QSAR prediction: migrated from IUCLID 5.6
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
QSAR model
Principles of method if other than guideline:
The acute toxicity to aquatic invertebrates was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
no
Details on test solutions:
not applicable
Test organisms (species):
Daphnia sp.
Details on test organisms:
not applicable
Test type:
other: QSAR
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
48h-EC50
Post exposure observation period:
not applicable
Hardness:
The QSAR is based on data from studies performed at acceptable hardness to ensure control survival
Test temperature:
The QSAR is based on data from studies performed at between 14 - 25 °C (depending on the species considered)
pH:
The QSAR is based on data from studies performed at acceptable pH between 6.0 - 9.0
Dissolved oxygen:
The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%)
Salinity:
The QSAR is based on data from studies performed on freshwater species
Nominal and measured concentrations:
The QSAR is based on data from studies performed using measured concentrations or with acceptable stability
Details on test conditions:
not applicable
Reference substance (positive control):
no
Remarks:
QSAR model
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.56 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL [0.33-0.95]
Details on results:
not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
95% CL [0.33-0.95]
QSAR statistical parameters are given in the QMRF and the QPRF

no data

Validity criteria fulfilled:
yes
Remarks:
The substance falls into the applicability domains of the QSAR model.
Conclusions:
48h-EC50 for camphene = 0.56 mg test item/L with 95% Confidence Limit between 0.33 and 0.95 mg test item/L.
Executive summary:

The acute toxicity to daphnia was determined using a validated QSAR for the Mode of Action in question, based on validated data derived from 48-hour EC50 tests on daphnia, for which the concentrations of the test item had been determined by chemical analyses over the test period. The QSAR is a simple linear regression of sub-cooled liquid solubility versus 48-h EC50 for daphnia and is valid within the applicability domain defined in the QMRF.

The acute toxicity to daphnia of camphene has been investigated using a QSAR model that predicts daphnia mortality in an OECD 202 study. Camphene falls within the applicability domain of the model as demonstrated in the QPRF.

The 48-h EC50 was 0.57 mg test material/L with 95%-Confidence Limit between 0.34 and 0.94 mg test material/L. EC50 were based on mobility. 

Based on the results of this study, Camphene would be classified as acute 1 to aquatic organisms in accordance with the classification of the CLP.

This toxicity study is acceptable and can be used for that endpoint.

 

Results Synopsis

Test Type: QSAR model

LC50: 0.56 mg test material/L 95%-C.L.: 0.33 to 0.95 mg test material/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
calculation (if not (Q)SAR)
Remarks:
Migrated phrase: estimated by calculation
Adequacy of study:
key study
Study period:
2014-05-23 to 2014-05-26
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Calculation method is used ; calculation method applicable for the endpoint
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
calculation method
Principles of method if other than guideline:
The acute toxicity to daphnia was determined using a validated QSAR for the Mode of Action in question. The first step of the iSafeRat mixture toxicity calculation employs phase equilibrium thermodynamics in order to determine the concentrations of each constituent within the WAF. This fraction equates to the analyzable fraction of a WAF study.
Within the WAF, the constituents also partition between themselves further reducing the bioavailable fraction and thus the toxicity of the mixture compared to the individual constituents. In the calculation the second step is to remove this non-bioavailable fraction.
The final step is to determine the truly bioavailable fraction of the WAF per constituent. The EC50s of each constituent are already known from literature or calculated using the iSafeRat QSAR model. Each value and calculation has been included as a supporting study in the IUCLID. An additivity approach (based on Chemical Activity of each constituent) is used in order to calculate the Effective Loading rate of the WAF.
The method has been validated using data derived from 48-hour EC50 tests on aquatic invertebrates, for which the concentrations of the test item had been determined by chemical analyses over the test period. Further to this the effective loading rate of the WAF is determined by using a series of calculation steps using phase equilibrium thermodynamics and excluding the non-bioavailable fraction.
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not applicable
Analytical monitoring:
no
Details on sampling:
not applicable
Vehicle:
no
Details on test solutions:
not applicable
Test organisms (species):
Daphnia sp.
Details on test organisms:
not applicable
Test type:
other: calculation method
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
48h-EL50 (effective loading rate of WAF)
Post exposure observation period:
not applicable
Hardness:
Hardness is not a necessary component of the WAF calculation
Test temperature:
The Temperature is not a necessary component of the WAF calculation but extremely low or high temperatures could influence the solubility of certain constituents. Therefore, the calculation method is considered acceptable to determine EL50s for daphnia between 12 and 28°C.
pH:
The pH is not a necessary component of the WAF calculation
Dissolved oxygen:
The oxygen concentration is not a necessary component of the WAF calculation
Salinity:
Salinity is not a necessary component of the WAF calculation. However as the fish QSAR for the constituents calculation was based on data from freshwater studies, the resulting calculation is considered valid for freshwater organisms
Nominal and measured concentrations:
The calculation determines measured concentrations
Details on test conditions:
calculation method
Reference substance (positive control):
not required
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
2.9 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: Typical mixture (Composition 1)
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
2.5 mg/L
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: Theoretical worst case mixture (Composition 2)
Details on results:
not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
not applicable

Table 1: Expected concentrations at this 48h-EL50 (composition 1: typical composition)

 

constituents

concentration in the WAF (mg.L-1)

limonene

0.20

α-pinene

1.26

β-pinene

0.47

Δ-3-carene

0.39

myrcene

0.10

camphene

0.04

bornyl acetate

0.05

β-phellandrene

0.05

Table 2: Expected concentrations at this 48h-EL50 (composition 2: theoretical worst case composition)

 

constituents

concentration in the WAF (mg.L-1)

limonene

0.16

α-pinene

1.38

β-pinene

0.60

Δ-3-carene

0.25

myrcene

0.04

camphene

0.03

bornyl acetate

0.03

β-phellandrene

0.0

Validity criteria fulfilled:
yes
Conclusions:
48h-EL50 for typical composition of Pine scotch oil = 2.9 mg test item /L and 48h-EL50 for theoretical worst case composition of Pine scotch oil = 2.5 mg test item/L
Executive summary:

Pine scotch oil is a Natural Complex Substance (UVCB) with a well-defined composition for which the relative percentage and its reported variation of each constituent is known. Its acute toxicity to aquatic invertebrate property has been investigated using an in-house calculation method that replaces an OECD 202 study and guideline for Testing of Chemicals No. 23 (i.e. WAF conditions).

Two theoretical compositions have been investigated, the “typical” composition proposed by the Lead registrant and a theoretical worst case composition that maximizes the concentration of the most toxic constituents.The acute toxicity to aquatic invertebrate was determined using a calculation method for Mode of Action 1 (non-polar narcotics) for each individual constituent present in the mixture.

 

The first step of the iSafeRat mixture toxicity calculation employs phase equilibrium thermodynamics in order to determine the concentrations of each constituent within the WAF. This fraction equates to the analysable fraction of a WAF study. In the calculation the second step is to remove this non-bioavailable fraction. Within the WAF, the constituents also partition between themselves further reducing the bioavailable fraction and thus the toxicity of the mixture compared to the individual constituents.

These two reasons explain why ecotoxicity values from WAF studies are always higher for non-polar narcotic mixtures than the calculated values from CLP additivity calculation.

The final step is to determine the truly bioavailable fraction of the WAF per constituent. The EC50s of each constituent are already known from literature or predicted using the iSafeRat QSAR model. Each value has been included as a supporting study in the IUCLID. An additivity approach (based on Chemical Activity of each constituent) is used in order to calculate the Effective Loading rate of the WAF.

The 48-h EL50 was 2.9 mg test material/L for the typical composition of Pine scotch oil, and the 48-h EL50 was 2.5 mg test material/L for the theoretical worst case composition. EL50 were based on mortality. 

Based on the results of this study, Pine scotch oil would not be classified as acute 1 to aquatic organisms in accordance with the classification of the CLP.

This toxicity study is acceptable and can be used for that endpoint.

 

Results Synopsis

Test Type: Calculation method

EL50: 2.9 mg test material/L for typical composition

EL50: 2.5 mg test material/L for theoretical worst case composition

Description of key information

Pine scotch oil is a Natural Complex Substance (UVCB) with a well-defined composition for which the relative percentage and its reported variation of each constituent is known. The acute toxicity to aquatic invertebrates property has been investigated using an in-house calculation method that mimics an OECD 202 study and guideline for Testing of Chemicals No. 23 (i.e. WAF conditions). The acute toxicity to aquatic invertebrates was determined using a calculation method for Mode of Action 1 (non-polar narcotics) for each individual constituent present in the mixture. This algorithm is based on a QSAR model which has been validated to be compliant with the OECD recommendations for QSAR modeling (OECD, 2004). Further to this the lethal loading rate of the WAF is determined by using a series of calculation steps using phase equilibrium thermodynamics first to determine the analyzable fraction (the concentration which should be analyzable in a WAF study) and then excluding the non-bioavailable fraction of the remaining constituents. The remaining, bioavailable fraction corresponds to the lethal loading value of the mixture. This approach has been validated using data derived from 48-hour EL50 tests on daphnia (OECD 202 study and the OECD guidance document on toxicity testing for difficult substances and mixtures No. 23, i.e. WAF conditions). Two theoretical compositions have been investigated, the “typical” composition proposed by the Lead registrant and a theoretical worst case composition that maximizes the concentration of the most toxic constituents. The 48-h EL50 was 2.9 mg test material/L for the typical composition of Pine scotch oil, and the 48-h EL50 was 2.5 mg test material/L for the theoretical worst case composition. EL50 were based on mortality.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
2.5 mg/L

Additional information

Pine scotch oil is a Natural Complex Substance (UVCB) with a well-defined composition for which the relative percentage of each constituent is known. Therefore, it has been decided that the ecotoxicity of Pine scotch oil will be derived from knowledge of the constituents; constituents approach.  The mixture ecotoxicity properties may be derived from the ecotoxicity of the individual constituents using the CLP additivity calculation approach.

However, CLP additivity approach is calculated on the basis that all the substances are at their maximum solubility and it has been observed that CLP additivity calculations for mode of action 1 compounds are unreasonably conservative when compared to classic WAF studies. This has been proved in a number of cases for natural complex substances. Indeed, natural extract compositions are a mixture of hydrophilic alcohol molecules and hydrophobic terpene molecules. Therefore, when a WAF is performed most of substances fully dissolved in the aqueous phase are the hydrophilic fraction while the hydrophobic fraction (the more toxic elements for MOA 1 substances) may be below their water solubility value.

The acute toxicity to aquatic invertebrate property for Pine scotch oil has been investigated using an in-house calculation method (iSafeRat® WAF module for mixture Toxicity calculation) that mimics an OECD 202 study and guideline for Testing of Chemicals No. 23 (i.e. WAF conditions).

Two theoretical compositions have been investigated, the “typical” composition proposed by the Lead registrant and a theoretical worst case composition that maximizes the concentration of the most toxic constituents. The acute toxicity to aquatic invertebrate was determined using a calculation method for Mode of Action 1 (non-polar narcotics) for each individual constituent present in the mixture.

 

The first step of the iSafeRat mixture toxicity calculation employs phase equilibrium thermodynamics in order to determine the concentrations of each constituent within the WAF. This fraction equates to the analysable fraction of a WAF study. In the calculation the second step is to remove this non-bioavailable fraction. Within the WAF, the constituents also partition between themselves further reducing the bioavailable fraction and thus the toxicity of the mixture compared to the individual constituents.

These two reasons explain why ecotoxicity values from WAF studies are always higher for non-polar narcotic mixtures than the calculated values from CLP additivity calculation.

The final step is to determine the truly bioavailable fraction of the WAF per constituent. The EC50s of each constituent are already known from literature or predicted using the iSafeRat QSAR model. Each value has been included as a supporting study in the IUCLID. An additivity approach (based on Chemical Activity of each constituent) is used in order to calculate the Effective Loading rate of the WAF.

 

 

Using this approach, the 48-h EL50 for fish was 2.9 mg test material/L for the typical composition of Pine scotch oil, and the 48-h EL50 was 2.5 mg test material/L for the theoretical worst case composition. These EL50 were based on mortality. 

Validation of iSafeRat® WAF module is presented in the attached document to the dossier. iSafeRat® Calculation remains conservative compared to the experimental value.

Based on the results of this study, Pine scotch oil would not be classified as acutely toxic to aquatic organisms in accordance with the classification of the CLP.

This toxicity prediction has been validated and is considered acceptable to fulfil the aquatic invertebrate toxicity endpoint.