Benzenamine, N-phenyl-, reaction products with styrene and 2,4,4-trimethylpentene

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not specified

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Data taken from HPV Challenge Program website from which data have been entered into the HPVIS: January 15, 2002 (original submission), October 23, 2004 and January 19, 2007.

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPPTS 850.1035 (Mysid Acute Toxicity Test)

Deviations:

yes

Remarks:

see below.

Principles of method if other than guideline:

The methodology as stated in the OECD SIDS documentation is listed as "OECD Guideline 471; B14 in EC Directive 92/69/EEC". This appears to be a typographical error in the data set, as this methodology relates to the Bacterial Reverse Mutation Assay. As such, it is assumed given the location of the study laboratory conducting the test, and the methodology followed, that the method is the US method for Mysid Acute Toxicity testing. This is therefore detailed above as the appropriate method.

GLP compliance:

yes

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:

Nominal and Measured Concentrations: 0, 1.3, 2.2, 3.6, 6, and 10 mg/l

Vehicle:

no

Details on test solutions:

Solutions of the test material were prepared by dilution with sea water. After mixing and allowing undissolved material to settle, the water soluble faction was added to two corresponding replicate test vessels.

Test organisms (species):

other:

Details on test organisms:

No further data specified within the report.

Test type:

flow-through

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No post exposure period specified within the report.

Hardness:

Not specified.

Test temperature:

Not specified.

pH:

Not specified.

Dissolved oxygen:

Not specified.

Salinity:

Not specified

Nominal and measured concentrations:

Not specified.

Details on test conditions:

Test Type: Flow-through
Test Vessel: No Data
Water Media Type: Saltwater
Test Concentrations: Unknown
Nominal and Measured Concentrations: 0, 1.3, 2.2, 3.6, 6, and 10 mg/l
Exposure Period: 96 Hours
Two control vessels were established containing the same dilution water but no test material. The test concentrations 0, 1.3, 2.2, 3.6, 6, and 10 mg/l were selected based on preliminary test results. Mysids then were added to the test and control vessels. Test organisms were carefully transferred into the appropriate concentrations of newly prepared vessels at the 24, 48, and 72 hour observation periods. In life observations and water analyses were conducted at 0, 24, 48, 72 and 96 hours.

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

< 1.3 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mortality

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.3 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mortality

Details on results:

Following 96-hours of exposure, 60-100% mortality was observed in the three highest concentrations, and 25 and 40% mortality was observed in the two lowest concentrations. The 96-hour LC50 was determined to be 2.3 mg/l (1.3-10 mg/l; 95% confidence limits. The 96-hour NOEC was < 1.3 mg/l.

Results with reference substance (positive control):

Not specified.

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 was determined to be 2.3 mg/l. The 96-hour NOEC was < 1.3 mg/l.

Executive summary:

Data taken from HPV Challenge Program website from which data have been entered into the HPVIS: January 15, 2002 (original submission), October 23, 2004 and January 19, 2007.

Following 96-hours of exposure, 60-100% mortality was observed in the three highest concentrations, and 25 and 40% mortality was observed in the two lowest concentrations. The 96-hour LC50 was determined to be 2.3 mg/l (1.3-10 mg/l; 95% confidence limits. The 96-hour NOEC was < 1.3 mg/l.  

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1035 (Mysid Acute Toxicity Test)

Deviations:

not specified

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Melting point: <0 °C
- Boiling point: 258 °C
- Vapour pressure: Not measured
- Water solubility (under test conditions): Not measured
OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: 8 % after 28 day
- Other:

Analytical monitoring:

no

Details on sampling:

Test Levels: Control, 150, 250, 400, 600 and 1,000 mg/L nominal test concentrations.

Vehicle:

no

Details on test solutions:

The test was conducted using the water accommodated fraction (WAF) of nominal test concentrations. Individual WAFs were prepared by adding a measured weight of test material to a measured volume of dilution water (1-L) in a glass vessel and stirring for 24 hours. Following the mixing period, the test solutions were allowed to stand for 1 hour before the water phase was siphoned off. The siphoned water phase (i.e., WAF) was used for the aquatic toxicity test.

Test organisms (species):

Americamysis bahia (previous name: Mysidopsis bahia)

Details on test organisms:

Test species: Juvenile mysids less than 24-hours old were produced from laboratory in-house culture.

Test type:

semi-static

Water media type:

saltwater

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation period specified.

Hardness:

Not specified

Test temperature:

Test Temperature (ºC) - 24 + 1

pH:

pH - 8.1

Dissolved oxygen:

TOC - 3.9 to 8.2

Salinity:

Water was adjusted to a salinity of 20 parts per thousand and aerated

Nominal and measured concentrations:

Nominal test concentrations

Details on test conditions:

A 2-L glass beaker that contained 1 L of test solution was used per treatment. The test vessels were loosely covered to reduce entry of dust, etc. Mysids were fed newly hatched Artemia salina nauplii once or twice daily during the test.
Dilution water: Seawater collected from the Atlantic Ocean in Hampton, New Hampshire was used. Water was adjusted to a salinity of 20 parts per thousand and aerated.
Water was free of pesticides and PCBs at the detection limit.
The WAF was used for testing. 10 mysids per test vessel (2 replicates per test concentration were used).

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

ca. 250 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 733 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Mysids exposed to 600 mg/L were lethargic and exhibited erratic swimming from 48 to 96 hours. No other sub lethal effects were observed in any test vessel during the 96 hour exposure. Effect concentrations based on nominal loading rates.

Results with reference substance (positive control):

Control response was satisfactory (>90% survival and no sublethal effects).

Reported statistics and error estimates:

EL50’s were calculated using standard statistical methods from Stephan (1983).

Validity criteria fulfilled:

not specified

Conclusions:

NOEC: c = 250 mg/l
EC50: c = 733 mg/l

Executive summary:

Study conducted to OECD test guidelines in compliance with GLP.Results: Effect concentrations based on nominal loading rates. Control response was satisfactory (>90% survival and no sublethal effects). Mysids exposed to 600 mg/L were lethargic and exhibited erratic swimming from 48 to 96 hours. No other sub lethal effects were observed in any test vessel during the 96 hour exposure. NOEC: c = 250 mg/l

EC50: c = 733 mg/l

Read across to supporting substance, CAS No. 36878 -20 -3 by structural analogue.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study, which is rated as reliability 2 because it is a read-across study.

Justification for type of information:

Please refer to IUCLID Section 13 for the read-across justification.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1035 (Mysid Acute Toxicity Test)

Deviations:

not specified

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Melting point: <0 °C
- Boiling point: 258 °C
- Vapour pressure: Not measured
- Water solubility (under test conditions): Not measured
OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: 8 % after 28 day
- Other:

Analytical monitoring:

no

Details on sampling:

Test Levels: Control, 150, 250, 400, 600 and 1,000 mg/L nominal test concentrations.

Vehicle:

no

Details on test solutions:

The test was conducted using the water accommodated fraction (WAF) of nominal test concentrations. Individual WAFs were prepared by adding a measured weight of test material to a measured volume of dilution water (1-L) in a glass vessel and stirring for 24 hours. Following the mixing period, the test solutions were allowed to stand for 1 hour before the water phase was siphoned off. The siphoned water phase (i.e., WAF) was used for the aquatic toxicity test.

Test organisms (species):

Americamysis bahia (previous name: Mysidopsis bahia)

Details on test organisms:

Test species: Juvenile mysids less than 24-hours old were produced from laboratory in-house culture.

Test type:

semi-static

Water media type:

saltwater

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation period specified.

Hardness:

Not specified

Test temperature:

Test Temperature (ºC) - 24 + 1

pH:

pH - 8.1

Dissolved oxygen:

TOC - 3.9 to 8.2

Salinity:

Water was adjusted to a salinity of 20 parts per thousand and aerated

Nominal and measured concentrations:

Nominal test concentrations

Details on test conditions:

A 2-L glass beaker that contained 1 L of test solution was used per treatment. The test vessels were loosely covered to reduce entry of dust, etc. Mysids were fed newly hatched Artemia salina nauplii once or twice daily during the test.
Dilution water: Seawater collected from the Atlantic Ocean in Hampton, New Hampshire was used. Water was adjusted to a salinity of 20 parts per thousand and aerated.
Water was free of pesticides and PCBs at the detection limit.
The WAF was used for testing. 10 mysids per test vessel (2 replicates per test concentration were used).

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

ca. 250 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 733 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Mysids exposed to 600 mg/L were lethargic and exhibited erratic swimming from 48 to 96 hours. No other sub lethal effects were observed in any test vessel during the 96 hour exposure. Effect concentrations based on nominal loading rates.

Results with reference substance (positive control):

Control response was satisfactory (>90% survival and no sublethal effects).

Reported statistics and error estimates:

EL50’s were calculated using standard statistical methods from Stephan (1983).

Validity criteria fulfilled:

not specified

Conclusions:

NOEC: c = 250 mg/l
EC50: c = 733 mg/l

Executive summary:

Study conducted to OECD test guidelines in compliance with GLP.Results: Effect concentrations based on nominal loading rates. Control response was satisfactory (>90% survival and no sublethal effects). Mysids exposed to 600 mg/L were lethargic and exhibited erratic swimming from 48 to 96 hours. No other sub lethal effects were observed in any test vessel during the 96 hour exposure. NOEC: c = 250 mg/l

EC50: c = 733 mg/l

Read across to supporting substance, CAS No. 36878 -20 -3 by structural analogue.

Description of key information

Assessment of acute toxicity to aquatic invertebrates.

Key value for chemical safety assessment

Marine water invertebrates

Marine water invertebrates

Effect concentration:

2.3 mg/L

Additional information

ANNEX XI - General Rules for Adaptation of the Standard Testing Regime set out in Annexes VII to X, Section 1. “Testing Does Not Appear Scientifically Necessary”, paragraph 1.1.2 states that:

 

Data shall be considered to be equivalent to data generated by the corresponding test methods referred to in Article 13(3) if the following conditions are met:

 

1) adequacy for the purpose of classification and labelling and/or risk assessment;

 

2) adequate and reliable coverage of the key parameters foreseen to be investigated in the corresponding test methods referred to in Article 13(3);

 

3) exposure duration comparable to or longer than the corresponding test methods referred to in Article 13(3) if exposure duration is a relevant parameter; and

 

4) adequate and reliable documentation of the study is provided.

 

The substancehas been supported under Environmental Protection Agency’s (EPA’s) High Production Volume (HPV) Challenge Program. The American Chemical Councils RAPA Panel, has derived a “Substituted Diphenylamines” category of chemicals, and assessed all available data.  To address the acute toxicity to aquatic invertebrates, two saltwater toxicity tests, using mysid shrimp are available; one on the substance itself, and one on a close structural analogue. The results of these are as follows:

 

CAS 68921-45-9: 96-h LC50 : 2.3 mg/l (1.3-10 mg/l; 95% confidence limits). 96-h NOEC < 1.3 mg/l.  

 

CAS No. 36878-20-3: 96 h EC50: c = 733 mg/l ; 96-h NOEC: c = 250 mg/l (WAF).

It is noted that the preferred study to address this endpoint utilises freshwater daphnia. However, the available data on saltwater species is considered appropriate to address the requirements of Annex XI, Para 1.1.2 as follows:

1) adequacy for the purpose of classification and labelling and/or risk assessment;

The two results available are somewhat different; this is attributed to the dosing methods seen in the experiments. For the purposes of hazard assessment, the value on the substance itself it taken as appropriate, and classification and labelling is applied on this basis. This is therefore considered adequate for risk assessment.

 

2) adequate and reliable coverage of the key parameters foreseen to be investigated in the corresponding test methods referred to in Article 13(3);

The studies are GLP compliant studies, conducted to recent guidelines. The integrity and results are therefore considered appropriate and are in accordance with the recognised guidelines. There is no documented evidence that saltwater species are more sensitive than freshwater species, although the REACH guidance attributes and additional assessment factor of 10 for risk characterisation calculations.

 

3) exposure duration comparable to or longer than the corresponding test methods referred to in Article 13(3) if exposure duration is a relevant parameter; and

The exposure duration of the study is in accordance with the guideline method at 96 hours. This is longer than the required 48 hour exposure utilised in the acute toxicity to Daphnia study.

 

4) adequate and reliable documentation of the study is provided.

The studies are GLP compliant studies, and have been peer reviewed and evaluated in the context of the OECD SIDS and US HP programme. The results are presented as appropriate summaries of the original reports.

 

The use of these studies results in classification and labelling under the CLP-Regulation (EC) No 1272/2008 as follows:

 

Aquatic Chronic Category 2

 

H411: Toxic to aquatic life with long lasting effects.