3-nitrotoluene

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Justification for type of information:

Due to insufficient reporting of the experimental details, this study was rated with a Klimisch score of 4 (‘not assignable’). As none of the available studies can serve as a key study with a sufficiently detailed reporting justifying a Klimisch score of 1 or 2 for the given endpoint, several independent sources were used in a weight of evidence approach. ‘Using weight of evidence implies that no single study of sufficient quality and reliability exists, and that information from several independent sources is required to be able to reach a conclusion on a particular property of the substance.' (guidance document How to prepare registration and PPORD dossiers, p. 87)

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): 3-nitrotoluene
- Analytical purity: > 98%

Test organisms (species):

Chlorella pyrenoidosa

Test type:

not specified

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

200 mg/l as CaCO3

Test temperature:

20 ± 0.5

pH:

8.4 ± 0.1

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

14 mg/L

Conc. based on:

test mat.

Basis for effect:

cell number

Remarks on result:

other: 95%

Results are given in the original reference as log EC50: log EC50 = 2.01 (EC50 µmol/l).
The concentrations causing 50 % reduction of the maximum density (yield)of Chlorella pyrenoidosa during a 96 h period of exposure were calculated according to Kooyman et al. (1983). Parametric analysis of population growth in bio-assays. Water Research 17: 527 - 538

Validity criteria fulfilled:

not specified

Executive summary:

A test on the acute toxicity of 3-nitrotoluene to the alga Chorella pyrenoidosa was performed according to the OECD Guideline 201 (Alga, Growth Inhibition Test). For a test period of 96 hours an EC50 value of 14 mg/l was reported (Deneer, 1988, 1989).

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Original report not available and documentation insufficient for assessment; exposure period too short: 6h

Justification for type of information:

Due to methodological deficiencies, this study was rated with a Klimisch score of 3 (‘not reliable’). As none of the available studies can serve as a key study with a sufficiently detailed reporting justifying a Klimisch score of 1 or 2 for the given endpoint, several independent sources were used in a weight of evidence approach. ‘Using weight of evidence implies that no single study of sufficient quality and reliability exists, and that information from several independent sources is required to be able to reach a conclusion on a particular property of the substance.’ (guidance document How to prepare registration and PPORD dossiers, p. 87). Despite being considered not  reliable (Klimisch 3), the experimental value obtained in this study still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

Principles of method if other than guideline:

only secondary source; method not detailed

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): 3-nitrotoluene
- Analytical purity: not reported

Test organisms (species):

Chlorella vulgaris

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

6 h

Duration:

6 h

Dose descriptor:

IC50

Effect conc.:

52 mg/L

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

Result is reported as log IC50 = 3.42 mol/l (corresponding to 52 mg/l).

Original report not available and documentation insufficient for assessment (data taken from literature but the original source remains unclear)

Validity criteria fulfilled:

not specified

Executive summary:

A test on the acute toxicity of 3-nitrotoluene to the alga Chorella pyrenoidosa was performed according in a static setup. For a test period of 6 hours an IC50 value of 52 mg/l was reported (Swann, 1995). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, the exposure time was too short: 6 h instead of the required 96h), the experimental value obtained in this study still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

Reference 3

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Documentation is insufficient for assessment. It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Justification for type of information:

Due to methodological deficiencies, this study was rated with a Klimisch score of 3 (‘not reliable’). As none of the available studies can serve as a key study with a sufficiently detailed reporting justifying a Klimisch score of 1 or 2 for the given endpoint, several independent sources were used in a weight of evidence approach. ‘Using weight of evidence implies that no single study of sufficient quality and reliability exists, and that information from several independent sources is required to be able to reach a conclusion on a particular property of the substance.’ (guidance document How to prepare registration and PPORD dossiers, p. 87). Despite being considered not  reliable (Klimisch 3), the experimental value obtained in this study still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

Principles of method if other than guideline:

other: cell multiplication inhibition test

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 3-nitrotoluol
- Analytical purity: no data

Analytical monitoring:

not specified

Test organisms (species):

Scenedesmus quadricauda

Details on test organisms:

- The concentration of the algal suspension of each test culture is measured turbidimetrically and expressed
as the extinction of the primary light of the monochromatic radiation at 578 nm for a 10 mm layer.

Test type:

not specified

Water media type:

freshwater

Limit test:

no

Total exposure duration:

8 d

Test temperature:

24 °C

pH:

7

Details on test conditions:

- Incubation of 10 ml test solution (algae in defined mineral medium with appropriate TS concentration)
- The test cultures prepared from the dilution series and the control cultures are kept under standardized
conditions for a period of 8 days, namely on a white surface protected against daylight and exposed
to constant lighting by luminescent tubes;

Duration:

8 d

Dose descriptor:

other: EC3

Effect conc.:

4.4 mg/L

Conc. based on:

test mat.

Basis for effect:

biomass

Reported TT (Toxicity threshold) is comparable to EC3; value refers to nominal TS concentration.

Validity criteria fulfilled:

not specified

Remarks:

It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Executive summary:

A test on the acute toxicity of 3-nitrotoluene to the alga Scenedesmus quadricauda was performed according in a cell multiplication inhibition test. For a test period of 8 days an EC3 value of 4.4 mg/l was reported (Bringmann, 1978-1980). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

Reference 4

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Documentation is insufficient for assessment. It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Justification for type of information:

Due to methodological deficiencies, this study was rated with a Klimisch score of 3 (‘not reliable’). As none of the available studies can serve as a key study with a sufficiently detailed reporting justifying a Klimisch score of 1 or 2 for the given endpoint, several independent sources were used in a weight of evidence approach. ‘Using weight of evidence implies that no single study of sufficient quality and reliability exists, and that information from several independent sources is required to be able to reach a conclusion on a particular property of the substance.’ (guidance document How to prepare registration and PPORD dossiers, p. 87). Despite being considered not  reliable (Klimisch 3), the experimental value obtained in this study still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

Principles of method if other than guideline:

Cell multiplication inhibition test

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 3-nitrotoluene
- Analytical purity: purity is not specified

Test organisms (species):

Microcystis aeruginosa

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

8 d

Duration:

8 d

Dose descriptor:

other: TT

Effect conc.:

1 mg/L

Conc. based on:

test mat.

Basis for effect:

other: cell count

It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Validity criteria fulfilled:

not specified

Remarks:

It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Executive summary:

A test on the acute toxicity of 3-nitrotoluene to the alga Microcystis aeruginosa was performed according in a static setup. For a test period of 8 days a TT (toxicity threshold, comparable to EC3) value of 1 mg/l was reported (Bringmann, 1978). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

Reference 5

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Documentation is insufficient for assessment. It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Justification for type of information:

Due to methodological deficiencies, this study was rated with a Klimisch score of 3 (‘not reliable’). As none of the available studies can serve as a key study with a sufficiently detailed reporting justifying a Klimisch score of 1 or 2 for the given endpoint, several independent sources were used in a weight of evidence approach. ‘Using weight of evidence implies that no single study of sufficient quality and reliability exists, and that information from several independent sources is required to be able to reach a conclusion on a particular property of the substance.’ (guidance document How to prepare registration and PPORD dossiers, p. 87). Despite being considered not  reliable (Klimisch 3), the experimental value obtained in this study still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

Principles of method if other than guideline:

Cell multiplication inhibition test

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 3-nitrotoluene
- Analytical purity: purity is not specified

Test organisms (species):

Scenedesmus quadricauda

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

8 d

Duration:

8 d

Dose descriptor:

other: TT

Effect conc.:

4.4 mg/L

Conc. based on:

test mat.

Basis for effect:

other: cell count

Reported TT (toxicity threshold) refers to nominal concentration and was determined at 3 % effect compared to the control (comparable to EC3)

 

Validity criteria fulfilled:

not specified

Remarks:

It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Executive summary:

A test on the acute toxicity of 3-nitrotoluene to the alga Scenedesmus quadricauda was performed in a static setup. For a test period of 8 days a TT (toxicity threshold, comparable to EC3) value of 4.4 mg/l was reported (Bringmann, 1977). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

Reference 6

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Documentation is insufficient for assessment. It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Justification for type of information:

Due to methodological deficiencies, this study was rated with a Klimisch score of 3 (‘not reliable’). As none of the available studies can serve as a key study with a sufficiently detailed reporting justifying a Klimisch score of 1 or 2 for the given endpoint, several independent sources were used in a weight of evidence approach. ‘Using weight of evidence implies that no single study of sufficient quality and reliability exists, and that information from several independent sources is required to be able to reach a conclusion on a particular property of the substance.’ (guidance document How to prepare registration and PPORD dossiers, p. 87). Despite being considered not  reliable (Klimisch 3), the experimental value obtained in this study still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

Principles of method if other than guideline:

other: Cell multiplication inhibition test

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 3-Nitrotoluol
- Analytical purity: 80 %

Analytical monitoring:

no

Test organisms (species):

Microcystis aeruginosa

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

8 d

Test temperature:

27° C

pH:

7

Details on test conditions:

- Algae were submitted to continuous artificial light
- Before starting the test, the stock solution of o-toluidine was neutralised.
- The solutions with the test concentrations were then prepared in 300 ml Erlenmeyer flasks.
- The extinction value of the monochromatic radiation (578 nm) of the cell suspension was
measured for each concentration of the test and control preparations.

Duration:

8 d

Dose descriptor:

other: EC3

Effect conc.:

1 mg/L

Basis for effect:

biomass

Reported TT (toxicity threshold) refers to nominal concentration and was determined at 3 % effect compared to the control (comparable to EC3).

Validity criteria fulfilled:

not specified

Remarks:

It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.

Executive summary:

A test on the acute toxicity of 3-nitrotoluene to the alga Microcystis aeruginosa was performed according in a static setup. For a test period of 8 days an EC3 value of 1 mg/l was reported (Bringmann, 1975, 1978). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

Reference 7

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Justification for type of information:

Due to insufficient reporting of the experimental details, this study was rated with a Klimisch score of 4 (‘not assignable’). As none of the available studies can serve as a key study with a sufficiently detailed reporting justifying a Klimisch score of 1 or 2 for the given endpoint, several independent sources were used in a weight of evidence approach. ‘Using weight of evidence implies that no single study of sufficient quality and reliability exists, and that information from several independent sources is required to be able to reach a conclusion on a particular property of the substance.' (guidance document How to prepare registration and PPORD dossiers, p. 87)

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): 3-nitrotoluene
- Analytical purity: > 98 %
- Molecular weight: 137.14

Analytical monitoring:

no

Details on test solutions:

The stock solution of the test substance was prepared in dimethylsulfoxide (DMSO, Merck, Purity 99%)

Test organisms (species):

Chlorella pyrenoidosa

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

14 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: reduction of the maximum density (yield)

Remarks on result:

other: 95% CL; 10-19

Validity criteria fulfilled:

not specified

Executive summary:

A test on the acute toxicity of 3-nitrotoluene to the alga Chorella pyrenoidosa was performed according to the OECD Guideline 201 (Alga, Growth Inhibition Test). For a test period of 96 hours an EC50 value of 14 mg/l was reported (Maas-Diepeveen and van Leeuwen, 1986).

Description of key information

In order to fulfill the data requirements a weight of evidence (WoE) approach (REACh Regulation (EC) No. 1907/2006, Annex XI Section 1.2) was chosen. This approach is a possible adaptation to the standard information requirements which ‘should be undertaken to maximise the use of existing data and minimise the commissioning of new in vivo testing’ (ECHA Guidance R.7a, 2017, p. 364). The WoE approach serves to reduce or avoid animal testing, which should only be performed as last resort (REACh, article 25). The individual study reports were conducted in accordance with standardized guidelines. The study reports nevertheless show some deficiencies with respect to the comprehensiveness of the presented experimental details. Therefore, a reliability of 1 or 2 and thus adequacy as key study was not assignable for any of the individual studies. Consequently, the studies were assigned a Klimisch score of 4 (according to guidance document R.4, ‘not assignable: studies or data […] which do not give sufficient experimental details […]). Collectively, these experimental studies can be used to conclude on this endpoint and to satisfy the information requirement.

A test on the acute toxicity of 3-nitrotoluene to the alga Chorella pyrenoidosa was performed according to the OECD Guideline 201 (Alga, Growth Inhibition Test). For a test period of 96 hours an EC50 value of 14 mg/l was reported (Maas-Diepeveen and van Leeuwen, 1986).

A test on the acute toxicity of 3-nitrotoluene to the alga Chorella pyrenoidosa was performed according to the OECD Guideline 201 (Alga, Growth Inhibition Test). For a test period of 96 hours an EC50 value of 14 mg/l was reported (Deneer, 1988, 1989).

A test on the acute toxicity of 3-nitrotoluene to the alga Chorella pyrenoidosa was performed according in a static setup. For a test period of 6 hours an IC50 value of 52 mg/l was reported (Swann, 1995). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, the exposure time was too short: 6 h instead of the required 96h), the experimental value obtained in this study still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

A test on the acute toxicity of 3-nitrotoluene to the alga Microcystis aeruginosa was performed according in a static setup. For a test period of 8 days an EC3 value of 1 mg/l was reported (Bringmann, 1975, 1978). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

A test on the acute toxicity of 3-nitrotoluene to the alga Microcystis aeruginosa was performed according in a static setup. For a test period of 8 days a TT (toxicity threshold, comparable to EC3) value of 1 mg/l was reported (Bringmann, 1978). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

A test on the acute toxicity of 3-nitrotoluene to the alga Scenedesmus quadricauda was performed according in a cell multiplication inhibition test. For a test period of 8 days an EC3 value of 4.4 mg/l was reported (Bringmann, 1978-1980). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

A test on the acute toxicity of 3-nitrotoluene to the alga Scenedesmus quadricauda was performed in a static setup. For a test period of 8 days a TT (toxicity threshold, comparable to EC3) value of 4.4 mg/l was reported (Bringmann, 1977). This result has been included as additional information. Despite being considered not  reliable (Klimisch 3, It is unclear whether the algae are within the exponential growth throughout the whole exposure period of 8 days.), the experimental value obtained in this study still supports the WoE conclusion.

Conclusion

The 96h-EC50 value of 14 mg/L reported independently by Maas-Diepeveen 1986 and Deneer 1988/1989 was chosen as key value for chemical safety assessment. This value is supported by other studies (TT/EC3/IC50-values in the range of: 1-52 mg/l).

Key value for chemical safety assessment

EC50 for freshwater algae:

14 mg/L