4-nitrotoluene

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:

other information

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

abstract

Justification for type of information:

Due to accesibility of only the abstract, 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 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

- Analytical method: HPLC, - Frequency of analysis: at the start and the end of exposure

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Test temperature:

20 ± 1°C

Nominal and measured concentrations:

Nominal and measured concentrations: 10.0, 5.56, 3.09, 1.71, 0.953 mg/L (geometric ratio: 1.8), control

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

4.27 mg/L

95% CI:

3.72 - 4.9

Nominal / measured:

nominal

Conc. based on:

test mat.

Validity criteria fulfilled:

not specified

Executive summary:

MITI, 1998

Effect concentrations

Species: Daphnia magna
Exposure duration : 48 h
Endpoint : EC50
Effect concentration : 4.27 mg/L
Basis for concentration : nominal
Remarks (e.g. 95% CL) : 95 % CL: 3.72 - 4.90 mg/L

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

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 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

Version: 1984

Deviations:

no

GLP compliance:

not specified

Specific details on test material used for the study:

- Analytical purity: not specified

Analytical monitoring:

yes

Test organisms (species):

Daphnia magna

Test type:

semi-static

Water media type:

freshwater

Remarks:

tap water filtrated by activated carbon (pH7.4~7.9, Ca/Mg=3.5~4.l, Na/K=5.5~6.4, hardness 20~32)

Total exposure duration:

48 h

Test temperature:

20 ± 1 °C

pH:

7.4-7.9

Dissolved oxygen:

8-9 ppm

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

5.46 mg/L

Remarks on result:

other: ± 0.37 (CV=6.8)

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

4.71 mg/L

Remarks on result:

other: ± 0.28 (CV=6.0)

Details on results:

CV= coefficient of variation
It is not clearly indicated in the results table, whether the reported results for 3-nitrotoluene were obtained with or without surfactant.

Validity criteria fulfilled:

not specified

Executive summary:

Kodoma, 2003

The toxicity of 4-nitrotoluene to the aquatic invertebrate Daphnia magna was evaluated in a 48 h study performed in accordance with OECD TG 202 (1984). The 48h-EC50 value was determined to be 4.71 mg/L and the 48h-LC50 value was reported as 5.46 mg/L. It is not clearly indicated in the results table, whether the reported results for 3-nitrotoluene were obtained with or without surfactant.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Basic data given. Restrictions of the method are: - Test period was only of 4 hours (represents only 8 % of the in the OECD and other current guidelines suggested test period) - Temperature during the test was higher (25 °C) than the one suggested by OECD and other current guidelines (22 °C) - Yeast is not a standard food for Daphnia Apparently Daphnia(?) data omitted in the publication process (Table 3 of the publication of Yen J-H, Lin K-H, Wang Y-S (2002) Acute lethal toxicity of environmental pollutants to aquatic organisms. Ecotoxicol Environ Safety 52: 113 - 116)

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: see below test conditions

GLP compliance:

not specified

Specific details on test material used for the study:

purity > 98 % (Purchased from E. Merck, Darmstadt, Germany)

Analytical monitoring:

not specified

Test organisms (species):

Daphnia pulex

Test type:

static

Water media type:

freshwater

Total exposure duration:

4 h

Hardness:

215 mg/l as CaCO3

Test temperature:

25 +/- 1 °C

pH:

6.6

Dissolved oxygen:

4.9 mg/L

Details on test conditions:

- Daphnia were fed with yeast in a 224 (diameter)x 46 (height)-cm circular plastic pool;
- acclimatization in aquaria for 2 weeks under conditions similar to those under which the test performed;

- 20 daphnia (24 hr after hatching) introduced in each 250 ml beaker containing 100 ml of test chemicals with 6 different concentrations (0, 50, 100, 200, 500, 1000 µg/ml) were prepared in duplicate
- mortality was observed after an incubation time of 4 h

Duration:

4 h

Dose descriptor:

LC50

Effect conc.:

41 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mobility

In Table 4 of their publication, Yen et al. report the LC50 to be 0.407 mg/l. After request, one of the authors (Wang 2003) communicated that the reported values are below the observed values by a factor of 100. Thus, the correct LC50 is 41 mg/l.

Validity criteria fulfilled:

not specified

Remarks:

Restrictions of the method: - Test period only 4 instead of 48h -Temperature during the test was higher (25 °C) than 22 °C - Yeast is not a standard food for Daphnia

Executive summary:

Yen, 2002, Wang, 2003

In Table 4 of their publication, Yen et al. report the LC50 for Daphnia magna to be 0.407 mg/l. After request, one of the authors (Wang 2003) communicated that the reported values are below the observed values by a factor of 100. Thus, the correct LC50 is 41 mg/l. Despite being considered not  reliable (Klimisch 3, Restrictions of the method: - Test period only 4 instead of 48h -Temperature during the test was higher (25 °C) than 22 °C - Yeast is not a standard food for Daphnia), 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 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Original report not available; Method not validated; test conducted for 24 hs (exposure time too short)

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.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Cited as Directive 84/449/EEC, C.2 (Acute toxicity for Daphnia)

GLP compliance:

not specified

Analytical monitoring:

no

Test organisms (species):

Daphnia magna

Test type:

not specified

Water media type:

freshwater

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

3 - 4 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

not specified

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

7 - 11 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

not specified

Validity criteria fulfilled:

not specified

Executive summary:

Hoechst AG, 1982

The acute toxicity of the test item 4-nitrotoluene towards the invertebrate Daphnia magna was tested according to Directive 84/449/EEC, C.2 (Acute toxicity for Daphnia). The following effect concentration was observed: EC50 (24h): 7-11 mg/L. Despite being considered not  reliable (Klimisch 3, the exposure time was too short: 24 h instead of the required 48h), 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 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

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: method not specified

GLP compliance:

not specified

Test organisms (species):

Daphnia magna

Measured EC50 concentrations were obtained from Aquire Database. They were compared with the predicted LC50 by using QSAR-models.
The duration of the test to determine LC50 and other details about the test system are not given.
LC50 measured for Daphnia magna: 11.0 mg/l

Validity criteria fulfilled:

not specified

Executive summary:

Fiedler, 1990 (presumably citing Bringmann, 1977)

Measured EC50 concentrations were obtained from Aquire Database. They were compared with the predicted LC50 by using QSAR-models. The duration of the test to determine LC50 and other details about the test system are not given.
LC50 measured for Daphnia magna: 11.0 mg/l. 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.

Reference 6

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

According to guideline study with acceptable restrictions, insufficient documentation

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)

Principles of method if other than guideline:

Method: other: comparable to OECD 202 part I (Daphnia, Acute Toxicity, 1984)

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in publication): 4-Nitrotoluene
- Analytical purity: no data

Analytical monitoring:

not specified

Test organisms (species):

other: Daphnia carinata

Details on test organisms:

- 6-24 hours old Daphnia were used for the test
- Daphnia were not fed during the test

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

22 +/-1 °C

Details on test conditions:

- photoperiod of 14 hours daylight/ 10 hours darkness
- 10 Daphnia in 25 mL of test water
- 60 Daphnia for each test

Duration:

48 h

Dose descriptor:

IC50

Effect conc.:

ca. 14.4 mg/L

Basis for effect:

mobility

Details on results:

Results are given as log IC50: log IC50 = 3.98 (IC50 mol/l)

Validity criteria fulfilled:

yes

Remarks:

validity criteria: - the results were considered valid if dissolved oxigen measured at the end of the test was at least equal to 60% of saturation, and if percentage of immobilisation observed for the controls was zero.

Executive summary:

Zhao, 1997

The toxicity of 4-nitrotoluene to the aquatic invertebrate Daphnia carinata was evaluated in a 48 h static test comparable to OECD 202 part I (Daphnia, Acute Toxicity, 1984). The 48h-IC50 (mobility) was determined to be 14.4 mg/L. This result has been included as additional information.

Reference 7

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Test procedure in accordance with national standard methods, without detailed documentation, but basic data given

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)

Principles of method if other than guideline:

other: NEN 6501: Determination of acute toxicity with Daphnia magna (1980) with slight modifications (Van Leeuwen et al. 1985)

GLP compliance:

not specified

Specific details on test material used for the study:

Name of test material (as cited in publication): 4-nitrotoluene
Analytical purity: > 98%

Analytical monitoring:

no

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Hardness:

200 mg/l CaCO3

Test temperature:

20 ± 0.5 °C

pH:

8.4 ± 0.1

Dissolved oxygen:

medium was saturated with air prior to use

Details on test conditions:

- During the tests daphnids were fed with Chlorella pyrenoidosa, which at the start of the experiments were present at a concentration of 1.0E+8 cells/L
- test was carried out in a room that was illuminated 12 h/day
- a synthetic test medium was used (NPR 6503,1980)

Duration:

48 h

Dose descriptor:

IC50

Effect conc.:

19 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mobility

Remarks on result:

other: 95% CL: (10-32)

Details on results:

Deneer et al. (1989) report the IC50 value as log IC50: log IC50 = 2.14 (IC50 µmol/L).

Reported statistics and error estimates:

Maas-Diepeveen and van Leeuwen (1986) calculated LC-50 48-h values and their 95 % confidence intervals according to Litchfield and Wilcoxon (1949).

Validity criteria fulfilled:

yes

Remarks:

- The oxygen content of all solutions did not decrease below 7.9 mg/l (85%) - Mortality in the controls never exceeded 10 %

Executive summary:

Maas-Diepeveen, 1986, Deneer, 1989

A test on the acute toxicity of 4-nitrotoluene to the invertebrate Daphnia magna was performed according to the Dutch Standardization Organization Method NEN 6501. For a test period of 48 hours an IC50 value of 19 mg/l (based on mobility) was reported.

Reference 8

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Test procedure according to national standards with some restrictions, limited documentation

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)

Principles of method if other than guideline:

other: Method by US-EPA 1975 (EPA-660/3-75-009)
The test was performed according to the procedure as described in the guideline method; with the exception that the temperature was maintained at 20 °C.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in publication): 4-Nitrotoluene
- Analytical purity: no data

Analytical monitoring:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

SRI stock culture

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

26 ppm (as CaCO3)

Test temperature:

20 °C

pH:

7.2-8.6

Details on test conditions:

alkalinity: 45 ppm (as CaCO3)
conductivity: 103 µmhos/cm

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

12.1 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mobility

Reported statistics and error estimates:

Effect values were calculated with the probit analysis according to Finney (1971)

Validity criteria fulfilled:

yes

Remarks:

The test was performed according to the procedure as described in the guideline method; with the exception that the temperature was maintained at 20 °C.

Executive summary:

Pearson, 1979

The acute toxicity of the test item 4-nitrotoluene towards the invertebrate Daphnia magna was tested in a static setup according to the method by US-EPA 1975 (EPA-660/3-75-009) (the test temperature of 20°C was the only exception of this guideline). The following effect concentration was observed: EC50 (48h): 12.1 mg/L.

Reference 9

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: BUA report

Adequacy of study:

weight of evidence

Reliability:

other: BUA report

Rationale for reliability incl. deficiencies:

other: no reliability is given as this is a summary entry for the BUA report

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)

Principles of method if other than guideline:

other: BUA report

GLP compliance:

not specified

Test organisms (species):

other: BUA report

Remarks on result:

other: key results see 'executive summary'

BUA report (1989):

 

The "Landesamt für Wasser und Abfall Nordrhein-Westfalen" reported the following p-nitrotoluene concentration as the threshold for effects on Daphnia magna: 11 mg/l (Schriftliche Mitteilung, 29.05.1979).

Bringmann & Kuehn (1982) found a 24h-EC0 of 5.6 mg/L, a 24h-EC50 of 9 mg/L and a 24h-EC100 of 12 mg/L (all immobility) for Daphnia magna. Liu et al. (1983) gives an 48h-LC50 (mortality) of 11.8 mg/L for the same species. Canton et al. (1985) measured a value of 7.5 mg/L as a 24h-LC50 and 24h-EC50 (immobilisation) for Daphnia magna.

For the oligochaeta Tubifex sp., p-nitrotoluene has shown a 24-h LC50 of 110 mg/l and a 48-h LC50 of 36 -64 mg/L (Yoshioka et al., 1986).

For two other species Canton et al. (1985) reports the following data for the acute toxicity (mortality): 48-h LC50 = 65 mg/L (Aedes aegypti), 48-h LC50 = 21 mg/L (Lymnaea stagnalis).

The hatching rate of Lymnaea stagnalis was reported to have a 7d-NOEC of 10 mg/L (Slooff & Canton, 1983).

Validity criteria fulfilled:

not specified

Executive summary:

BUA report (1989):

 

The "Landesamt für Wasser und Abfall Nordrhein-Westfalen" reported the following p-nitrotoluene concentration as the threshold for effects on Daphnia magna: 11 mg/l (Schriftliche Mitteilung, 29.05.1979).

Bringmann & Kuehn (1982) found a 24h-EC0 of 5.6 mg/L, a 24h-EC50 of 9 mg/L and a 24h-EC100 of 12 mg/L (all immobility) for Daphnia magna. Liu et al. (1983) gives an 48h-LC50 (mortality) of 11.8 mg/L for the same species. Canton et al. (1985) measured a value of 7.5 mg/L as a 24h-LC50 and 24h-EC50 (immobilisation) for Daphnia magna.

For the oligochaeta Tubifex sp., p-nitrotoluene has shown a 24-h LC50 of 110 mg/l and a 48-h LC50 of 36 -64 mg/L (Yoshioka et al., 1986).

For two other species Canton et al. (1985) reports the following data for the acute toxicity (mortality): 48-h LC50 = 65 mg/L (Aedes aegypti), 48-h LC50 = 21 mg/L (Lymnaea stagnalis).

The hatching rate of Lymnaea stagnalis was reported to have a 7d-NOEC of 10 mg/L (Slooff & Canton, 1983).

Reference 10

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

Method not validated; test conducted for 24 hs

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: Immobilization test according to Bringmann & Kuehn (1977)

GLP compliance:

no

Specific details on test material used for the study:

Name of the material (as cited in publication): 4-Nitrotoluol
Analytical purity: no data

Analytical monitoring:

no

Details on sampling:

no data

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

- The fastest profilerating clone of Daphnias was selected from about 30 clones isolated from a pond
- Age of daphnids at start of test: <= 24h

Test type:

static

Water media type:

freshwater

Total exposure duration:

24 h

Hardness:

286 mg CaCO3/L (=16 German degrees)

Test temperature:

20-22 °C

pH:

Initial pH 7.6 - 7.7 (not adjusted during exposure)

Details on test conditions:

- Test vessels: 50 mL breakers with 20 mL filling volume
- Water media type: Chlorine free tap water
- The Daphnias were fed cultured Chlorella vulgaris daily
- For incubation 10 Daphnias per concentration were used
- Triplicate samples per concentration
- Test vessels loosely capped with filter paper

Duration:

24 h

Dose descriptor:

LC0

Effect conc.:

7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

LC100

Effect conc.:

20 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

11 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Validity criteria fulfilled:

not specified

Executive summary:

A test on the acute toxicity of 4-nitotoluene to the invertebrate Daphnia magna was performed (immobilization test according to Bringmann & Kuehn (1977)). For a test period of 24 hours an LC50 value of 11 mg/l was reported. Despite being considered not  reliable (Klimisch 3, the exposure time was too short: 24 h instead of the required 48h), 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 (Bringmann, 1977).

Reference 11

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: OECD SIDS

Adequacy of study:

weight of evidence

Reliability:

other: OECD SIDS

Rationale for reliability incl. deficiencies:

other: no reliability is given as this is a summary entry for the OECD SIDS

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)

Principles of method if other than guideline:

OECD SIDS

GLP compliance:

not specified

Test organisms (species):

other: OECD SIDS

Remarks on result:

other: key results see 'executive summary'

OECD SIDS (2003):

 

Several studies on the toxicity of 4-nitrotoluene to invertebrates were summarized in the assessment of the OECD SIDS (2003).

Zhao et al. (1995) and Zhao & Wang (1995) obtained in a test according to OECD Guideline 202 part I an acute toxicity (24 h-EC50) of 6.4 mg/L (nominal concentration). In a test conducted in analogy to the OECD 202 proposal of 1979 a 48 h-EC50 of 7.5 mg/l was determined. The results of the stability experiment performed before testing Daphnia toxicity showed a decay of the test compound in the test medium of only 6 % after 8 days. Therefore nominal concentrations can be used (Canton et al., 1985). A test according to the US-EPA method described in EPA-660/375-009 in 1975 showed a 48 h-EC50 of 11.8 mg/l (Liu et al., 1983). With Daphnia pulex Yen and coworkers (2002) obtained an 4h-EC50 of 41 mg/l. Endpoint in all these studies was immobilisation.

Validity criteria fulfilled:

not specified

Executive summary:

OECD SIDS (2003):

Several studies on the toxicity of 4-nitrotoluene to invertebrates were summarized in the assessment of the OECD SIDS (2003).

Zhao et al. (1995) and Zhao & Wang (1995) obtained in a test according to OECD Guideline 202 part I an acute toxicity (24 h-EC50) of 6.4 mg/L (nominal concentration). In a test conducted in analogy to the OECD 202 proposal of 1979 a 48 h-EC50 of 7.5 mg/l was determined. The results of the stability experiment performed before testing Daphnia toxicity showed a decay of the test compound in the test medium of only 6 % after 8 days. Therefore nominal concentrations can be used (Canton et al., 1985). A test according to the US-EPA method described in EPA-660/375-009 in 1975 showed a 48 h-EC50 of 11.8 mg/l (Liu et al., 1983). With Daphnia pulex Yen and coworkers (2002) obtained an 4h-EC50 of 41 mg/l. Endpoint in all these studies was immobilisation.

Reference 12

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Guideline study without detailed documentation, basic data given.

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:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Deviations:

not specified

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): 4-methylnitrobenzene
- purity: >=98 %

Analytical monitoring:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

Test organisms sieved to obtain animals less than 24 h old

Test type:

other: not specifed

Water media type:

freshwater

Total exposure duration:

48 h

Hardness:

Ca hardness of 250 +/- 25 mg/L obtained by addition of 294 mg/L CaCl2*2H2O, 123.3 mg/L MgSO4*7H2O, 65 mg/L NaHCO3, 5.8 mg/KCl

Test temperature:

20 +/- 2°C

pH:

7.8 +/- 0.2

Dissolved oxygen:

>90%

Details on test conditions:

- No food during incubation
- Each incubation vessel up to 20 Daphnias, at least 2 ml incubation medium per animal
- Daily check of oxygen concentration, pH, temperature
- Quality criteria:
- Oxygen concentration at least 2 mg/l
- Immobilization does not exceed 10 % in controls

Reference substance (positive control):

yes

Remarks:

K2Cr2O7

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 4.2 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mobility

Details on results:

Results are given in the original reference of Liu et al. (1997) as log LC50: log EC50 = 0.62 expressed in [mg/L].

Results with reference substance (positive control):

24 h-EC50: 0.6 - 1.7 mg/L

Reported statistics and error estimates:

Probit analysis of data

Validity criteria fulfilled:

yes

Remarks:

- Daily check of oxygen concentration, pH, temperature - Quality criteria: - Oxygen concentration at least 2 mg/l - Immobilization does not exceed 10 % in controls

Executive summary:

A test on the acute toxicity of 4-nitotoluene to the invertebrate Daphnia magna was performed according to ISO 6341 15. For a test period of 48 hours an EC50 value of ca. 4.2 mg/l was reported (Liu et al., 1997).

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.

Weight of Evidence (range of the 48h IC/LC50: 4.2 -19 mg/L)

A test on the acute toxicity of 4-nitotoluene to the invertebrate Daphnia magna was performed according to ISO 6341 15. For a test period of 48 hours an EC50 value of ca. 4.2 mg/l was reported (Liu et al., 1997).

 

OECD SIDS (2003): Several studies on the toxicity of 4-nitrotoluene to invertebrates were summarized in the assessment of the OECD SIDS (2003).

Zhao et al. (1995) and Zhao & Wang (1995) obtained in a test according to OECD Guideline 202 part I an acute toxicity (24 h-EC50) of 6.4 mg/L (nominal concentration). In a test conducted in analogy to the OECD 202 proposal of 1979 a 48 h-EC50 of 7.5 mg/l was determined. The results of the stability experiment performed before testing Daphnia toxicity showed a decay of the test compound in the test medium of only 6 % after 8 days. Therefore nominal concentrations can be used (Canton et al., 1985). A test according to the US-EPA method described in EPA-660/375-009 in 1975 showed a 48 h-EC50 of 11.8 mg/l (Liu et al., 1983). With Daphnia pulex Yen and coworkers (2002) obtained an 4h-EC50 of 41 mg/l. Endpoint in all these studies was immobilisation.

 

BUA report (1989):

The "Landesamt für Wasser und Abfall Nordrhein-Westfalen" reported the following p-nitrotoluene concentration as the threshold for effects on Daphnia magna: 11 mg/l (Schriftliche Mitteilung, 29.05.1979).

Bringmann & Kuehn (1982) found a 24h-EC0 of 5.6 mg/L, a 24h-EC50 of 9 mg/L and a 24h-EC100 of 12 mg/L (all immobility) for Daphnia magna. Liu et al. (1983) gives an 48h-LC50 (mortality) of 11.8 mg/L for the same species. Canton et al. (1985) measured a value of 7.5 mg/L as a 24h-LC50 and 24h-EC50 (immobilisation) for Daphnia magna.

For the oligochaeta Tubifex sp., p-nitrotoluene has shown a 24-h LC50 of 110 mg/l and a 48-h LC50 of 36 -64 mg/L (Yoshioka et al., 1986).

For two other species Canton et al. (1985) reports the following data for the acute toxicity (mortality): 48-h LC50 = 65 mg/L (Aedes aegypti), 48-h LC50 = 21 mg/L (Lymnaea stagnalis).

The hatching rate of Lymnaea stagnalis was reported to have a 7d-NOEC of 10 mg/L (Slooff & Canton, 1983).

 

A test on the acute toxicity of 4-nitrotoluene to the invertebrate Daphnia magna was performed according to the Dutch Standardization Organization Method NEN 6501. For a test period of 48 hours an IC50 value of 19 mg/l (based on mobility) was reported. (Maas-Diepeveen, 1986, Deneer, 1989)

 

The acute toxicity of the test item 4-nitrotoluene towards the invertebrate Daphnia magna was tested in a static setup according to the method by US-EPA 1975 (EPA-660/3-75-009) (the test temperature of 20°C was the only exception of this guideline). The following effect concentration was observed: EC50 (48h): 12.1 mg/L. (Pearson, 1979)

 

The toxicity of 4-nitrotoluene to the aquatic invertebrate Daphnia carinata was evaluated in a 48 h static test comparable to OECD 202 part I (Daphnia, Acute Toxicity, 1984). The 48h-IC50 (mobility) was determined to be 14.4 mg/L. This result has been included as additional information. (Zhao, 1997)

 

The toxicity of 4-nitrotoluene to the aquatic invertebrate Daphnia magna was evaluated in a 48 h study performed in accordance with OECD TG 202 (1984). The 48h-EC50 value was determined to be 4.71 mg/L and the 48h-LC50 value was reported as 5.46 mg/L. It is not clearly indicated in the results table, whether the reported results for 3-nitrotoluene were obtained with or without surfactant. (Kodoma, 2003)

MITI, 1998, effect concentrations

Species: Daphnia magna
Exposure duration : 48 h
Endpoint : EC50
Effect concentration : 4.27 mg/L
Basis for concentration : nominal
Remarks (e.g. 95% CL) : 95 % CL: 3.72 - 4.90 mg/L

 

The most conservative 48h-EC50 value of 4.2 mg/L (Liu, 1997) is chosen as key value for assessment.

Supporting studies

Despite being considered not  reliable (Klimisch 3, the exposure time was too short: 24 h instead of the required 48h), the experimental value obtained in these studies still supports the WoE conclusion confirming that the EC50 is in the order of magnitude of the concluded key value.

 

The acute toxicity of the test item 4-nitrotoluene towards the invertebrate Daphnia magna was tested according to Directive 84/449/EEC, C.2 (Acute toxicity for Daphnia). The following effect concentration was observed: EC50 (24h): 7-11 mg/L (Hoechst AG, 1982).

A test on the acute toxicity of 4-nitotoluene to the invertebrate Daphnia magna was performed (immobilization test according to Bringmann & Kuehn (1977)). For a test period of 24 hours an LC50 value of 11 mg/l was reported. (Bringmann, 1977)

Measured EC50 concentrations were obtained from Aquire Database. They were compared with the predicted LC50 by using QSAR-models. The duration of the test to determine LC50 and other details about the test system are not given.
LC50 measured for Daphnia magna: 11.0 mg/l. (Fiedler, 1990; presumably citing Bringmann, 1977)

In Table 4 of their publication, Yen et al. report the LC50 for Daphnia magna to be 0.407 mg/l. After request, one of the authors (Wang 2003) communicated that the reported values are below the observed values by a factor of 100. Thus, the correct LC50 is 41 mg/l. Despite being considered not  reliable (Klimisch 3, Restrictions of the method: - Test period only 4h instead of 48h -Temperature during the test was higher (25 °C) than 22 °C - Yeast is not a standard food for Daphnia), 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. (Yen, 2002, Wang, 2003)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

4.2 mg/L