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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 based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Study meets the criteria for Klimisch code 2 because it followed an internationally recognised method (EPA OTS 797.1300) and was carried out to Good Laboratory Practice. However, there was no analytical confirmation of the test concentrations. In addition, there were deviations from the guideline, but these deviations did not affect the study outcome.
Justification for type of information:
REPORTING FORMAT FOR THE CATEGORY APPROACH
1. HYPOTHESIS FOR THE CATEGORY APPROACH
The read across follows Scenario 5 - Qualitatively and quantitatively similar effects are caused by a common compound, which is formed from all category members (as described in the 2017 Read-Across Assessment Framework document).
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
TARGET: Benzenesulfonic acid, di-C10-14-alkyl derivs., sodium salts
SOURCE: Benzenesulphonic acid, C14-24 branched / linear alkyl derivative, calcium salt / Benzenesulphonic acid, C14-24 branched / linear alkyl derivative, calcium salt
3. CATEGORY APPROACH JUSTIFICATION

Linear and non-linear or branched alkylbenzene sulfonates are anionic surfactants with molecules characterized by a hydrophobic (apolar) and a hydrophilic (polar) group. As a group of chemicals, they are generally mixtures of closely related isomers and homologues. Each molecule contains an aromatic ring sulfonated at the para position and attached to either a linear or a branched alkyl chain at any position except the terminal carbons. The sulfonate group is a common functional group present in each of the category members, and is expected to exhibit similar biological activities with little influence from the length of carbon chain. The cation components of the chemicals (e.g. calcium, magnesium, sodium, or barium) are not expected to contribute significantly to the toxicity.
4. DATA MATRIX
See Read Across document attached to CSR
Qualifier:
according to
Guideline:
EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
yes
Remarks:
TOC in dilution water at the beginning of test was greater than 2 mg/L. Hardness was not determined in the water used for culture and acclimation. These deviations did not affect the outcome of the test.
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Total organic carbon (TOC) measurements of initial (0 hours) test solutions and at test termination (48 hours).
Vehicle:
no
Details on test solutions:
The 100, 300 and 1000 mg/L nominal loadings were prepared using sterilised filtered well water. Individual water accommodation fractions were prepared by stirring the vessel for 24 hours with a Teflon coated magnetic stir bar and removing the water phase for the toxicity test.

Dilution water was filtered well water collected at Hampton, New Hamshire. Water was adjusted to an appropriate hardness, passed through activated carbon and then an ultraviolet steriliser. Water was stored in a polyethylene tank where it was aerated. Dilution water had a total organic carbon concentration of 2.8 mg/L and a total suspended solids concentration of <10 mg/L.

No undissolved test material was reported on the surface of the test vessels during the entire toxicity test.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Waterflea
- Source: In-house laboratory culture
- Age at study initiation (mean and range, SD): Juvenile (<24 hours old)


ACCLIMATION
- Acclimation conditions: Daphnid cultures were maintained in 100% dilution water under static reneal conditions at 19.0 to 20.4°C. The dissolved oxygen concentration was always at least 8.0 mg/L
- Health during acclimation: During the 24 hours preceeding the test, daphnids were not treated for disease and were free from apparent sickness, injuries and abnormalities at the beginning of the test
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
190 mg CaCO3/L at the beginning of the test and 188 mg CaCO3/L at the end of the test
Test temperature:
Refer to Table 1
pH:
Refer to Table 2
Dissolved oxygen:
Refer to Table 3
Salinity:
Not relevant
Nominal and measured concentrations:
Control, 100, 300 and 1000 mg/L WAF.

TOC levels were 2.8-3.5 mg/L in the control, 2.8-3.6 mg/L at 100 mg/L WAF, 3.0-3.7 mg/L at 300 mg/L WAF and 2.7-3.4 mg/L at 1000 mg/L WAF loading. TOC levels were not considered to be indicative of actual test material concentrations and results are therefore based on nominal loading rates.
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL glass beaker containing 200 mL of test solution, loosely covered to reduce the entry of dust
- No. of organisms per vessel: Ten
- No. of vessels per concentration (replicates): Two

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Filtered well water collected at Hampton, New Hampshire
- Total organic carbon: Refer to Table 4
- Alkalinity: pH 7.2 to 8.1
- Conductivity: 860 to 880 µmhos/cm

OTHER TEST CONDITIONS
- Photoperiod: 16-hours light per day
- Light intensity: 20 µEin/m²

EFFECT PARAMETERS MEASURED: Immobilisation

TEST CONCENTRATIONS
- Test concentrations: Control, 100, 300 and 1000 mg/L nominal WAF
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Details on results:
Control immobilisation did not exceed the test validity criterion (i.e. <10% immobilisation in the controls).

At 24-hours, no immobilised or dead organisms were observed in the control or treatments. At 48-hours, 5, 0, 20 and 5% immobilisation were reported for control, 100, 300 and 1000 mg/L, respectively.
Reported statistics and error estimates:
The statistical analysis of survival data is not warranted.

Table 5. Survival and sub-lethal effect data

Concentration (mg/L)

Replicate

Number of survivors

No of affected Daphnia

0 hours

3 hours

24 hours

48 hours

0 hours

3 hours

24 hours

48 hours

0 (control)

1

10

10

10

9

0

0

0

0

2

10

10

10

10

0

0

0

0

100

1

10

10

10

10

0

0

0

0

2

10

10

10

10

0

0

0

0

300

1

10

10

10

7

0

0

0

0

2

10

10

10

9

0

0

0

0

1000

1

10

10

10

10

0

0

0

0

2

10

10

10

9

0

0

0

0

Validity criteria fulfilled:
yes
Conclusions:
The test substance was not toxic to daphnids at loading rates up to 1000 mg/L.
Executive summary:

The 48–hour acute toxicity of benzenesulphonic acid, C14-24 branched / linear alkyl derivative, calcium salt to Waterfleas (Daphnia magna) was studied under static conditions. Daphnids were exposed to control and test chemical at nominal concentrations of 100, 300 and 1000 mg/L WAF for 48 hours.  Mortality/immobilisation and sublethal effects were observed daily.

 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age: Neonates (<24 hours old)

Test Type: Static

Percent survival/unaffected organisms was 95% in the control, 100% at 100 mg/l, 80% at 300 mg/l and 95% at 1000 mg/l.

48 hour No Observed Effect Loading Rate (NOELR) = 1000 mg/l

48 hour Effective Loading Rate (EL50) = >1000 mg/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Study period:
1993
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Study meets the criteria for Klimisch score 2 because it followed an internationally recognised method (EPA OTS 797.1300) and was carried out to Good Laboratory Practice. However, there was no analytical confirmation of the test concentrations. In addition, there were deviations from the guideline, but these deviations did not affect the study outcome.
Justification for type of information:
REPORTING FORMAT FOR THE CATEGORY APPROACH
1. HYPOTHESIS FOR THE CATEGORY APPROACH
The read across follows Scenario 5 - Qualitatively and quantitatively similar effects are caused by a common compound, which is formed from all category members (as described in the 2017 Read-Across Assessment Framework document).
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
TARGET: Benzenesulfonic acid, di-C10-14-alkyl derivs., sodium salts
SOURCE: benzenesulfonic acid, mono C15-30 branched alkyl and di C11-13 branched and linear alkyl derivatives./ benzenesulfonic acid, mono C15-30 branched alkyl and di C11-13 branched and linear alkyl derivatives.
3. CATEGORY APPROACH JUSTIFICATION

Linear and non-linear or branched alkylbenzene sulfonates are anionic surfactants with molecules characterized by a hydrophobic (apolar) and a hydrophilic (polar) group. As a group of chemicals, they are generally mixtures of closely related isomers and homologues. Each molecule contains an aromatic ring sulfonated at the para position and attached to either a linear or a branched alkyl chain at any position except the terminal carbons. The sulfonate group is a common functional group present in each of the category members, and is expected to exhibit similar biological activities with little influence from the length of carbon chain. The cation components of the chemicals (e.g. calcium, magnesium, sodium, or barium) are not expected to contribute significantly to the toxicity.
4. DATA MATRIX
See Read Across document attached to CSR
Qualifier:
according to
Guideline:
EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
no
Remarks:
TOC in dilution water at the beginning of test was greater than 2 mg/L. Hardness was not determined in the water used for culture and acclimation. The vortex did not extend to 30-50% of the mixing vessel and the WAF settled for longer than 1 hour.
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Total organic carbon (TOC) measurements of initial (0 hours) test solutions and at test termination (48 hours)
Vehicle:
no
Details on test solutions:
The 100, 300 and 1000 mg/L nominal loadings were prepared using sterilised filtered well water. Individual water accommodation fractions were prepared by stirring the vessel for 24 hours with a Teflon coated magnetic stir bar and removing the water phase for the toxicity test.

Dilution water was filtered well water collected at Hampton, New Hamshire. Water was adjusted to an appropriate hardness, passed through activated carbon and then an ultraviolet steriliser. Water was stored in a polyethylene tank where it was aerated. Dilution water had a total organic carbon concentration of 2.3 mg/L and a total suspended solids concentration of <10 mg/L.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Waterflea
- Source: In-house laboratory culture
- Age at study initiation (mean and range, SD): Juvenile (<24 hours old)


ACCLIMATION
- Acclimation conditions: Daphnid cultures were maintained in 100% dilution water under static reneal conditions at 20.0 to 20.2°C. The dissolved oxygen concentration was always at least 8.8 mg/L
- Health during acclimation: During the 24 hours preceeding the test, daphnids were not treated for disease and were free from apparent sickness, injuries and abnormalities at the beginning of the test
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not reported.
Hardness:
180 mg CaCO3/L
Test temperature:
Refer to Table 1
pH:
Refer to Table 2
Dissolved oxygen:
Refer to Table 3
Salinity:
Not relevant
Nominal and measured concentrations:
Control, 100, 300 and 1000 mg/L WAF.

TOC levels were 2.3-2.8 mg/L in the control, 2.8-2.9 mg/L at 100 mg/L WAF, 2.6-2.8 mg/L at 300 mg/L WAF and 2.6-3.0 mg/L at 1000 mg/L WAF loading. TOC levels were not considered to be indicative of actual test material concentrations and results are therefore based on nominal loading rates.
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL glass beaker containing 200 mL of test solution, loosely covered to reduce the entry of dust
- No. of organisms per vessel: Ten
- No. of vessels per concentration (replicates): Two

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Filtered well water collected at Hampton, New Hampshire
- Total organic carbon: Refer to Table 4
- Alkalinity: pH 7.2 to 8.1
- Conductivity: Refer to Table 5

OTHER TEST CONDITIONS
- Photoperiod: 16-hours light per day
- Light intensity: 20 µEin/m²

EFFECT PARAMETERS MEASURED: Immobilisation

TEST CONCENTRATIONS
- Test concentrations: Control, 100, 300 and 1000 mg/L nominal WAF
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Details on results:
Control immobilisation did not exceed the test validity criterion (i.e. <10% immobilisation in the controls).

At 24-hours, no immobilisation or dead organisms were observed in the control or at 300 and 1000 mg/L WAF, but 5% immobilisation/mortality was seen in the 100 mg/L WAF treatment. At 48-hours, 0, 10, 0 and 0% immobilisation were reported for control, 100, 300 and 1000 mg/L WAF, respectively.
Reported statistics and error estimates:
The statistical analysis of survival data is not warranted.

Table 6. Survival and sublethal effect data

Concentration (mg/L)

Replicate

Number of survivors

No of floating Daphnia

0 hours

3 hours

24 hours

48 hours

0 hours

3 hours

24 hours

48 hours

0 (control)

1

10

10

10

10

0

0

0

0

2

10

10

10

10

0

0

0

0

100

1

10

10

9

9

0

10

7

6

2

10

10

10

9

0

10

10

10

300

1

10

10

10

10

0

0

0

3

2

10

10

10

10

0

0

0

3

1000

1

10

10

10

10

0

0

3

3

2

10

10

10

10

0

0

5

7

No sublethal effects were noted (floating was observed, but is not a sublethal effect)

Validity criteria fulfilled:
yes
Conclusions:
The test substance was not toxic to daphnids are loading rates up to 1000 mg/L.
Executive summary:

The 48–hour acute toxicity of benzenesulfonic acid, mono C15-30 branched alkyl and di C11-13 branched and linear alkyl derivatives, calcium salts, overbased to Waterfleas (Daphnia magna) was studied under static conditions. Daphnids were exposed to control and test chemical at nominal concentrations of 100, 300 and 1000 mg/L WAF for 48 hours. Mortality/immobilisation and sublethal effects were observed daily.

 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age: Neonates (<24 hours old)

Test Type: Static

Percent survival/unaffected organisms was 95% in the control, 100% at 100 mg/l, 80% at 300 mg/l and 95% at 1000 mg/l.

48 hour No Observed Effect Loading Rate (NOELR) = 1000 mg/L

48 hour Effective Loading Rate (EL50) = >1000 mg/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Study meets criteria for Klimisch code 2 because it followed an internationally recognised method (EPA OTS 797.1300) and was carried out to Good Laboratory Practice. However, there was no analytical confirmation of the test concentrations. In addition, there were deviations from the guideline, but these deviations did not affect the study outcome.
Justification for type of information:
REPORTING FORMAT FOR THE CATEGORY APPROACH
1. HYPOTHESIS FOR THE CATEGORY APPROACH
The read across follows Scenario 5 - Qualitatively and quantitatively similar effects are caused by a common compound, which is formed from all category members (as described in the 2017 Read-Across Assessment Framework document).
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
TARGET: Benzenesulfonic acid, di-C10-14-alkyl derivs., sodium salts
SOURCE: Benzenesulfonic acid, mono and dialkyl derivatives, magnesium salts
3. CATEGORY APPROACH JUSTIFICATION

Linear and non-linear or branched alkylbenzene sulfonates are anionic surfactants with molecules characterized by a hydrophobic (apolar) and a hydrophilic (polar) group. As a group of chemicals, they are generally mixtures of closely related isomers and homologues. Each molecule contains an aromatic ring sulfonated at the para position and attached to either a linear or a branched alkyl chain at any position except the terminal carbons. The sulfonate group is a common functional group present in each of the category members, and is expected to exhibit similar biological activities with little influence from the length of carbon chain. The cation components of the chemicals (e.g. calcium, magnesium, sodium, or barium) are not expected to contribute significantly to the toxicity.
4. DATA MATRIX
See Read Across document attached to CSR
Qualifier:
according to
Guideline:
EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
yes
Remarks:
TOC in dilution water at the beginning of test was greater than 2 mg/L. Hardness was not determined in the water used for culture and acclimation. The vortex did not extend to 30-50% of the mixing vessel.
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Total organic carbon (TOC) measurements of initial (0 hours) test solutions and at test termination (48 hours)
Vehicle:
no
Details on test solutions:
The 100, 300 and 1000 mg/L nominal loadings were prepared using sterilised filtered well water. Individual water accommodation fractions were prepared by stirring the vessel for 24 hours with a Teflon coated magnetic stir bar and removing the water phase for the toxicity test.

Dilution water was filtered well water collected at Hampton, New Hamshire. Water was adjusted to an appropriate hardness, passed through activated carbon and then an ultraviolet steriliser. Water was stored in a polyethylene tank where it was aerated. Dilution water had a total organic carbon concentration of 2.9 mg/L and a total suspended solids concentration of <10 mg/L.

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Waterflea
- Source: In-house laboratory culture
- Age at study initiation (mean and range, SD): Juvenile (<24 hours old)

ACCLIMATION
- Acclimation conditions: Daphnid cultures were maintained in 100% dilution water under static reneal conditions at 20.4 to 20.7°C. The dissolved oxygen concentration was always at least 8.6 mg/L
- Health during acclimation: During the 24 hours preceeding the test, daphnids were not treated for disease and were free from apparent sickness, injuries and abnormalities at the beginning of the test

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
176 mg CaCO3/L at the beginning of the test, 180 mg CaCO3/L at the end of the test
Test temperature:
Refer to Table 1
pH:
Refer to Table 2
Dissolved oxygen:
Refer to Table 3
Salinity:
Not relevant
Nominal and measured concentrations:
Control, 100, 300 and 1000 mg/L WAF.

TOC levels were 1.8-2.8 mg/L in the control, 2.2-3.3 mg/L at 100 mg/L WAF, 2.1-3.2 mg/L at 300 mg/L WAF and 1.8-3.3 mg/L at 1000 mg/L WAF loading. TOC levels were not considered to be indicative of actual test material concentrations and results are therefore based on nominal loading rates.
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL glass beaker containing 200 mL of test solution, loosely covered to reduce the entry of dust
- No. of organisms per vessel: Ten
- No. of vessels per concentration (replicates): Two

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Filtered well water collected at Hampton, New Hampshire
- Total organic carbon: Refer to Table 4
- Alkalinity: pH 7.0 to 8.4
- Conductivity: Refer to Table 5

OTHER TEST CONDITIONS
- Photoperiod: 16-hours light per day
- Light intensity: 9 µEin/m²

EFFECT PARAMETERS MEASURED: Immobilisation

TEST CONCENTRATIONS
- Test concentrations: Control, 100, 300 and 1000 mg/L nominal WAF
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Details on results:
Control immobilisation did not exceed the test validity criterion (i.e. <10% immobilisation in the controls).

No immobilisation of daphnids was observed in the control and all the treatments.
Reported statistics and error estimates:
The statistical analysis of survival data is not warranted.

Table 6. Survival and sublethal effect data

Concentration (mg/L)

Replicate

Number of survivors

No of affected

0 hours

3 hours

24 hours

48 hours

0 hours

3 hours

24 hours

48 hours

0 (control)

1

10

10

10

10

0

0

0

0

2

10

10

10

10

0

0

0

0

100

1

10

10

10

10

0

0

0

0

2

10

10

10

10

0

0

0

0

300

1

10

10

10

10

0

0

0

0

2

10

10

10

10

0

0

0

0

1000

1

10

10

10

10

0

0

0

0

2

10

10

10

10

0

0

0

0

Conclusions:
The test substance was not toxic to daphnids are loading rates up to 1000 mg/L.
Executive summary:

The 48–hour acute toxicity of benzenesulfonic acid.mon and dialkyl derivatives, magnesium salts to Waterfleas (Daphnia magna) was studied under static conditions. Daphnids were exposed to control and test chemical at nominal concentrations of 100, 300 and 1000 mg/L WAF for 48 hours. Mortality/immobilisation and sublethal effects were observed daily.

 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age: Neonates (<24 hours old)

Test Type: Static

No immobilisation of daphnids was observed in the control and all the treatments.

48 hour No Observed Effect Loading Rate (NOELR) = 1000 mg/L

48 hour Effective Loading Rate (EL50) = >1000 mg/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Study meets criteria for Klimisch code 2 because it followed an internationally recognised method (OECD Guideline 202) and was carried out to Good Laboratory Practice. However, there was no analytical confirmation of the test concentrations.
Justification for type of information:
REPORTING FORMAT FOR THE CATEGORY APPROACH
1. HYPOTHESIS FOR THE CATEGORY APPROACH
The read across follows Scenario 5 - Qualitatively and quantitatively similar effects are caused by a common compound, which is formed from all category members (as described in the 2017 Read-Across Assessment Framework document).
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
TARGET: Benzenesulfonic acid, di-C10-14-alkyl derivs., sodium salts
SOURCE: Alkyl benzene sulfonate salts of calcium, magnesium, sodium, or barium.
3. CATEGORY APPROACH JUSTIFICATION

Linear and non-linear or branched alkylbenzene sulfonates are anionic surfactants with molecules characterized by a hydrophobic (apolar) and a hydrophilic (polar) group. As a group of chemicals, they are generally mixtures of closely related isomers and homologues. Each molecule contains an aromatic ring sulfonated at the para position and attached to either a linear or a branched alkyl chain at any position except the terminal carbons. The sulfonate group is a common functional group present in each of the category members, and is expected to exhibit similar biological activities with little influence from the length of carbon chain. The cation components of the chemicals (e.g. calcium, magnesium, sodium, or barium) are not expected to contribute significantly to the toxicity.
4. DATA MATRIX
See Read Across document attached to CSR
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
no
Details on sampling:
- Concentrations: 0 and 100 mg/l nominal WAF loading rates
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: An amount of test material (250 mg) was added to the surface of 2.5 litres of reconstituted water in a 3 litre glass beaker. The test solution was stirred for 24 hours using a magnetic stirrer using a stirring rate such that a slight dimple was formed on the surface. After stirring the mixture was 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 5 cm for from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aquoous phase was removed by mid-depth siphoning (the first 75-100 ml discarded).
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Microscopic inspection of the WAF showed no micro-dispersions or undissolved test material to be present.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Waterflea
- Source: In-house laboratory cultures
- Age at study initiation: 1st instar
- Feeding during test: Daphnids were not fed during the test

ACCLIMATION
- Acclimation conditions: Adult Daphnia were maintained in polypropylene vessels containing approximately 2 litres of reconstituted water in a temperature controled room at approximately 20°C. Gravid adults were isolated tge 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.
- Lighting cycle: 16 hours light, 8 hours dark with 20 minute dawn and dusk transition periods
- Type and amount of food: Algae (Chlorella sp.)
- Feeding frequency: Each culture was fed daily with a suspension of algae (Chlorella sp.)
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
Approximate theoretical total hardness of 250 mg/l
Test temperature:
Refer to Table 1
pH:
Refer to Table 2
Dissolved oxygen:
Refer to Table 3
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 ml glass jar containing approximately 200 ml of test preparation loosely covered by aluminium foil to reduce evaporation
- No. of organisms per vessel: Five
- No. of vessels per concentration (replicates): Four
- No. of vessels per control (replicates): Four
- Biomass loading rate: Not reported


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water as described below


OTHER TEST CONDITIONS
- Photoperiod: 16 hours light, 8 hours dark with 20 minute dawn and dusk transition periods


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisatin and adverse reactions were recorded at 24 and 48 hours after the start of exposure. Daphnia were considered immobile if they were unable to swim for approximately 15 seconds after gentle agitation


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 10 and 100 mg/l nominal WAF loading rates
- Results used to determine the conditions for the definitive study: No immobilisation was observed at the 10 and 100 mg/l WAF loading rates
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:
other: WAF loading rate
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Details on results:
No immobilisation was observed in any Daphnids at 100 mg/l nominal WAF loading rate
Results with reference substance (positive control):
24 hour EC50 = 1.5 mg/l (95% CL = 1.4-1.7 mg/l)
48 hour EC50 = 1.2 mg/l (95% CL = 1.0-1.3 mg/l)
Reported statistics and error estimates:
The EC50s and associated confidence limits for the reference substance were calculated using the Spearman-Karber method
Executive summary:

The 48–hour acute toxicity of sulphonic acids, petroleum, calcium salts to Waterfleas (Daphnia magna) was studied under static conditions. Daphnids were exposed to control and test chemical at a nominal concentration of 100 mg/L WAF for 48 hours. Immobilisation and sublethal effects were observed daily.

 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age: <24 hours old

Test Type: Static

No immobilisation of daphnids was observed in the control and all the treatments.

48 hour No Observed Effect Loading Rate (NOELR) = 100 mg/L

48 hour Effective Loading Rate (EL50) = >100 mg/L

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Study meets criteria for Klimisch code 2 because it followed an internationally recognised method (OECD Guideline 202) and was carried out to Good Laboratory Practice. However, there was no analytical confirmation of the test concentrations.
Justification for type of information:
REPORTING FORMAT FOR THE CATEGORY APPROACH
1. HYPOTHESIS FOR THE CATEGORY APPROACH
The read across follows Scenario 5 - Qualitatively and quantitatively similar effects are caused by a common compound, which is formed from all category members (as described in the 2017 Read-Across Assessment Framework document).
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
TARGET: Benzenesulfonic acid, di-C10-14-alkyl derivs., sodium salts
SOURCE: Sulphonic acids, petroleum, sodium salts
3. CATEGORY APPROACH JUSTIFICATION

Linear and non-linear or branched alkylbenzene sulfonates are anionic surfactants with molecules characterized by a hydrophobic (apolar) and a hydrophilic (polar) group. As a group of chemicals, they are generally mixtures of closely related isomers and homologues. Each molecule contains an aromatic ring sulfonated at the para position and attached to either a linear or a branched alkyl chain at any position except the terminal carbons. The sulfonate group is a common functional group present in each of the category members, and is expected to exhibit similar biological activities with little influence from the length of carbon chain. The cation components of the chemicals (e.g. calcium, magnesium, sodium, or barium) are not expected to contribute significantly to the toxicity.
4. DATA MATRIX
See Read Across document attached to CSR
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
no
Details on sampling:
- Concentrations: 0 and 100 mg/l nominal WAF loading rates
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: An amount of test material (250 mg) was added, via syringe, to the surface of 2.5 litres of reconstituted water. The test solution was stirred for 24 hours using a magnetic stirrer using a stirring rate such that a slight dimple was formed on the surface. After stirring the mixture was 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 5 cm for from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aquoous phase was removed by mid-depth siphoning (the first 75-100 ml discarded).
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Microscopic inspection of the WAF showed no micro-dispersions or undissolved test material to be present
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Waterflea
- Source: In-house laboratory cultures
- Age at study initiation: 1st instar
- Feeding during test: Daphnids were not fed during the test

ACCLIMATION
- Acclimation conditions: Adult Daphnia were maintained in polypropylene vessels containing approximately 2 litres of reconstituted water in a temperature controled room at approximately 20°C. Gravid adults were isolated tge 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.
- Lighting cycle: 16 hours light, 8 hours dark with 20 minute dawn and dusk transition periods
- Type and amount of food: Algae (Chlorella sp.)
- Feeding frequency: Each culture was fed daily with a suspension of algae (Chlorella sp.)
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
Approximate theoretical total hardness of 250 mg/l
Test temperature:
Refer to Table 1
pH:
8.0
Dissolved oxygen:
Refer to Table 2
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 ml glass jar containing approximately 200 ml of test preparation loosely covered by aluminium foil to reduce evaporation
- No. of organisms per vessel: Five
- No. of vessels per concentration (replicates): Four
- No. of vessels per control (replicates): Four
- Biomass loading rate: Not reported


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water as described below


OTHER TEST CONDITIONS
- Photoperiod: 16 hours light, 8 hours dark with 20 minute dawn and dusk transition periods


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisatin and adverse reactions were recorded at 24 and 48 hours after the start of exposure. Daphnia were considered immobile if they were unable to swim for approximately 15 seconds after gentle agitation


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 10 and 100 mg/l nominal WAF loading rates in the initial range finding study, 0, 1.0 and 10 mg/l nominal WAF loading rates in the second range finding study and 0, 1.0, 10 and 100 mg/l in the third range finding study
- Results used to determine the conditions for the definitive study: Immobilisation was observed at 10 and 100 mg/l WAF loading rates in the first range finding study, therefore the second study was conducted. No immobilisation was observed at 1.0 and 10 mg/l WAF loading rates in the second trange finding study, therefore the third study was conducted. No immobilisation was observed at the 1.0, 10 and 100 mg/l WAF loading rates in the third study.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mobility
Details on results:
No immobilisation was observed in any Daphnids at 100 mg/l nominal WAF loading rate
Results with reference substance (positive control):
24 hour EC50 = 1.5 mg/l (95% CL = 1.4-1.7 mg/l)
48 hour EC50 = 1.2 mg/l (95% CL = 1.0-1.3 mg/l)
Reported statistics and error estimates:
The EC50s and associated confidence limits for the reference substance were calculated using the Spearman-Karber method
Executive summary:

The 48–hour acute toxicity of sulphonic acids, petroleum, sodium salts to Waterfleas (Daphnia magna) was studied under static conditions. Daphnids were exposed to control and test chemical at a nominal concentration of 100 mg/L WAF for 48 hours. Immobilisation and sublethal effects were observed daily.

 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age: <24 hours old

Test Type: Static

No immobilisation of daphnids was observed in the control and all the treatments.

48 hour No Observed Effect Loading Rate (NOELR) = 100 mg/L

48 hour Effective Loading Rate (EL50) = >100 mg/L

Description of key information

All data for short-term toxicity to invertebrates are from two studies on read across source substances were conducted according to OECD TG 202 or EPA OTS 797.1300. In each of these studies, the 48 hour Effective Loading Rate was equivalent to the highest concentrations tested.

 

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
100 mg/L

Additional information

Short-term toxicity to invertebrates studies on two read across source substances and conducted according to OECD TG 202 or EPA OTS 797.1300 are available. In each of these studies, the 48 hour Effective Loading Rate was equivalent to the highest concentrations tested. In short-term studies performed according to EPA OTS 797.1300 (Daphnid Acute Toxicity Test), water fleas (Daphnia magna) were exposed to four concentrations (0,100, 300, and 1000 mg/L) of test substance under a static conditions (Ward, 1993). Test substances were tested independently and include benzenesulphonic acid, C14-24 branched/ linear alkyl derivative, calcium salt (CASN 115733-09-0), benzenesulfonic acid, mono C15-30 branched alkyl and di C11-13 branched and linear alkyl derivatives, calcium salts, overbased (CASN 71486-79-8) or benzenesulfonic acid mono and dialkyl derivatives, magnesium salts (CASN 71786-47-5). Mortality/immobilization were observed daily. The 48-hour No Observed Effect Loading Rate (NOELR) was 1000 mg/L when 24 hour old daphnids were exposed in each of the studies. The 48 hour Effective Loading Rate (EL50) is >1000 mg/L based on nominal concentrations.

In another study performed according to OECD TG 202 (Daphnia sp. Acute Immobilisation Test), water fleas (Daphnia magna) were exposed to 0 and 100 mg/L of either sulfonic acids, petroleum, calcium salts (CASN 61789-86-4) or sulfonic acids, petroleum, sodium salts (CASN 68608-26-4) for a period of 48 hours under static conditions (Goodband, 2005). In each of the studies the 48 hour No Observed Effective Loading Rate (NOELR) was 100 mg/L and the 48 hour Effective Loading Rate (EL50) was >100 mg/L.