N-1-naphthylaniline

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:

documentation insufficient for assessment

Qualifier:

according to guideline

Guideline:

other: not specified

Test type:

not specified

Water media type:

not specified

Total exposure duration:

48 h

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.26 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

mortality

Validity criteria fulfilled:

not specified

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

48h acute bioassay using Daphnia magna as test organism (following recommendations of US-EPA)

GLP compliance:

not specified

Analytical monitoring:

no

Details on sampling:

not applicable

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: adding ethanol stock solutions of PNA to water
- Controls: yes, blank and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol (0.5 mL/L water)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: waterflea
- Source: biological supply house
- Age at study initiation (mean and range, SD): 12 +/- 12 hours, first instar
- Method of breeding: stock cultures with regular subculturing (method of Dewey and Parker), in 4L polyethylene separatory funnels
- Feeding during test
- Food type: no

ACCLIMATION
- Acclimation period: stock cultures with regular subculturing
- Acclimation conditions (same as test or not): yes
- Type and amount of food: baker's yeast, chlorophyll and algae (Chlorella pyrenoidosa)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

44 mg/L as CaCO3

Test temperature:

20 ± 2°C

pH:

7.2-7.6

Dissolved oxygen:

not specified

Nominal and measured concentrations:

Nominal: 0, 0.13, 0.22, 0.36, 0.60 and 1.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: culture dishes containing 100 mL test solution
- Aeration: no data
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: soft reconstituted freshwater, recommended by US EPA for culture and testing of freshwater invertebrates; reagent-grade, deionized water (> 10megohm-cm), filtered through activated carbon and a 0.2 µm particle filter in a Barnstead NANOpure water purification unit
- Alkalinity: 30-35 mg/L as CaCO3
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 16 h light: 8 h darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): immobilisation

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 0.25-0.35

Details on results:

- Mortality of control: blank control 5% (1/20), solvent control 15% (3/20) after 48 h

Reported statistics and error estimates:

LC50 values and their 95% confidence intervals were calculated by the moving average method of Thompson when the data permitted (constant dilution factor between the toxicant concentration). Otherwise, the LC50 was estimated by the method of Litchfield and Wilcoxon which requires at least two concentrations producing partial mortalities. If this method could not be used, the LC50 was estimated by graphical interpolation.

Cumulative mortalities:

0.13 mg/L:       0% after 24 hours, 5% after 48 hours

0.22 mg/L:       0% after 24 hours, 30% after 48 hours

0.36 mg/L:       0% after 24 hours, 55% after 48 hours

0.60 mg/L:       0% after 24 hours, 100% after 48 hours

1.00 mg/L:       50% after 24 hours, 90% after 48 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to Daphnia magna in a bioassay screening the acute toxicity (48h) of the test substance.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study following national guideline (of South Korea)

Qualifier:

according to guideline

Guideline:

other: Acute toxicity testing Daphnia magna of Notification No. 1997-9 of NIER adopted in 1997

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 82g of test substance was put in a flask of 10 mL volume and a drop of HCO-40 was included then, DMSO was added up to the demarcation lineto prepare solution with a concentration of 8000 mg/L. The solution was diluted for use. The study solution was prepared for each concentration by putting stock solution into the same amount of the diluted water.
- Controls: Only diluted water was used for control groups. solvent control group was prepared by putting a drop of HCO-40 into a flask of 10 mL and adding DMSO up to the demarcated line. The same amount used for the highest concnetration was put into the diluted water.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): HCO-40, DMSO
- Concentration of vehicle in test medium (stock solution and final test solution): not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: in-house culture
- Age at study initiation (mean and range, SD): < 24h
- Feeding during test: no

ACCLIMATION
- Acclimation period: not specified
- Acclimation conditions (same as test or not): same
- Type and amount of food: 10E5 cells/mL Scenedesmus subspicatus
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

174 mg/L as CaCO3

Test temperature:

20.1 - 20.7 °C

pH:

7.11- 7.31

Dissolved oxygen:

8.3 - 8.6 mg/L

Nominal and measured concentrations:

Nominal: 0.05, 0.1, 0.2, 0.4 and 0.8 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 150 mL crystallizing dish with 100 mL test solution
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- No. of vessels per vehicle control (replicates): 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to US-EPA guideline EPA/600/4-85/013 adopted in 1985
- Alkalinity: 114 mg/L as CaCO3
- Culture medium different from test medium: no
- Intervals of water quality measurement: Dissolved oxygen and pH were measured at the start and the end of the study.

OTHER TEST CONDITIONS
- Photoperiod: 16h light/ 8h dark
- Light intensity: 1840 - 2180 lux

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48h test duration.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.58 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 0.55 -0.62 mg/L

Details on results:

- Behavioural abnormalities: none
- Mortality of control: no
- Other adverse effects control: no

Validity criteria fulfilled:

not specified

Conclusions:

The test substance showed to be acutely very toxic to Daphnia magna in a bioassay screening the acute toxicity (48h) of the test substance.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

48h acute bioassay using Daphnia magna as test organism (following recommendations of US-EPA)

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: adding ethanol stock solutions of PNA to water
- Controls: yes, blank and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol (0.2 mL/L water)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: biological supply house
- Age at study initiation (mean and range, SD): 12 +/- 12 hours, first instar
- Method of breeding: stock cultures with regular subculturing (method of Dewey and Parker), in 4L polyethylene separatory funnels
- Feeding during test
- Food type: no

ACCLIMATION
- Acclimation period: stock cultures with regular subculturing
- Acclimation conditions (same as test or not): yes
- Type and amount of food: baker's yeast, chlorophyll and algae (Chlorella pyrenoidosa)

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

44 mg/L as CaCO3

Test temperature:

20 ± 2 °C

pH:

7.2-7.6

Dissolved oxygen:

not specified

Nominal and measured concentrations:

Nominal: 0, 0.08, 0.13, 0.22, 0.36, 0.60 and 1.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: culture dishes containing 100 mL test solution
- Aeration: no data
- Renewal rate of test solution (frequency/flow rate): animals were transferred to fresh solutions after 24 h of exposure
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: soft reconstituted freshwater, recommended by US EPA for culture and testing of freshwater invertebrates; reagent-grade, deionized water (> 10megohm-cm), filtered through activated carbon and a 0.2 µm particle filter in a Barnstead NANOpure water purification unit
- Alkalinity: 30-35 mg/L as CaCO3
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 16 h light: 8 h darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality (lack of movement)

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.67 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 0.50-0.90

Details on results:

- Mortality of control: blank control 0% (0/10), solvent control 10% (1/10) after 48 h

Reported statistics and error estimates:

LC50 values and their 95% confidence intervals were calculated by the moving average method of Thompson when the data permitted (constant dilution factor between the toxicant concentration). Otherwise, the LC50 was estimated by the method of Litchfield and Wilcoxon which requires at least two concentrations producing partial mortalities. If this method could not be used, the LC50 was estimated by graphical interpolation.

Cumulative mortalities:

0.08 mg/L:       0% after 24 and 48 hours

0.13 mg/L:       0% after 24 and 48 hours

0.22 mg/L:       0% after 24 and 48 hours

0.36 mg/L:       10% after 24 and 48 hours

0.60 mg/L:       30% after 24 hours, 40% after 48 hours

1.00 mg/L:       70% after 24 hours, 80% after 48 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to Daphnia magna in a bioassay screening the acute toxicity (48h) of the test substance.

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

48h acute bioassay using Daphnia magna as test organism (following recommendations of US-EPA)

GLP compliance:

not specified

Analytical monitoring:

no

Details on sampling:

not applicable

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: adding ethanol stock solutions of PNA to water
- Controls: yes, blank and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol (0.33 mL/L water)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Source: biological supply house
- Age at study initiation (mean and range, SD): 7 +/- 1 days, adults
- Method of breeding: stock cultures with regular subculturing (method of Dewey and Parker), in 4L polyethylene separatory funnels
- Feeding during test
- Food type: no

ACCLIMATION
- Acclimation period: stock cultures with regular subculturing
- Acclimation conditions (same as test or not): yes
- Type and amount of food: baker's yeast, chlorophyll and algae (Chlorella pyrenoidosa)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

44 mg/L as CaCO3

Test temperature:

20 ± 2 °C

pH:

7.2-7.6

Dissolved oxygen:

not specified

Nominal and measured concentrations:

Nominal: 0, 0.13, 0.22, 0.36, 0.60 and 1.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: culture dishes containing 100 mL test solution
- Aeration: no data
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: soft reconstituted freshwater, recommended by US EPA for culture and testing of freshwater invertebrates; reagent-grade, deionized water (> 10megohm-cm), filtered through activated carbon and a 0.2 µm particle filter in a Barnstead NANOpure water purification unit
- Alkalinity: 30-35 mg/L as CaCO3
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 16 h light: 8 h darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): immobilisation

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.68 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 0.53-0.88

Details on results:

- Mortality of control: blank control 10% (2/20), solvent control 5% (1/20) after 48 h

Reported statistics and error estimates:

LC50 values and their 95% confidence intervals were calculated by the moving average method of Thompson when the data permitted (constant dilution factor between the toxicant concentration). Otherwise, the LC50 was estimated by the method of Litchfield and Wilcoxon which requires at least two concentrations producing partial mortalities. If this method could not be used, the LC50 was estimated by graphical interpolation.

Cumulative mortalities:

0.13 mg/L:       5% after 24 hours, 5% after 48 hours

0.22 mg/L:       10% after 24 hours, 20% after 48 hours

0.36 mg/L:       0% after 24 hours, 5% after 48 hours

0.60 mg/L:       5% after 24 hours, 50% after 48 hours

1.00 mg/L:       0% after 24 hours, 65% after 48 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to Daphnia magna in a bioassay screening the acute toxicity (48h) of the test substance.

Reference 6

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

48h acute bioassay using Daphnia magna as test organism (following recommendations of US-EPA)

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: adding ethanol stock solutions of PNA to water
- Controls: yes, blank and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol (0.2 mL/L water)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Source: biological supply house
- Age at study initiation (mean and range, SD): 12 +/- 12 hours, first instar
- Method of breeding: stock cultures with regular subculturing (method of Dewey and Parker), in 4L polyethylene separatory funnels
- Feeding during test
- Food type: no

ACCLIMATION
- Acclimation period: stock cultures with regular subculturing
- Acclimation conditions (same as test or not): yes
- Type and amount of food: baker's yeast, chlorophyll and algae (Chlorella pyrenoidosa)

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

170 mg/L as CaCO3

Test temperature:

20 ± 2 °C

pH:

7.2-7.6

Dissolved oxygen:

not specified

Nominal and measured concentrations:

Nominal: 0, 0.08, 0.13, 0.22, 0.36, 0.60 and 1.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: culture dishes containing 100 mL test solution
- Aeration: no data
- Renewal rate of test solution (frequency/flow rate): animals were transferred to fresh solutions after 24 h of exposure
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: soft reconstituted freshwater, recommended by US EPA for culture and testing of freshwater invertebrates; reagent-grade, deionized water (> 10megohm-cm), filtered through activated carbon and a 0.2 µm particle filter in a Barnstead NANOpure water purification unit
- Alkalinity: 30-35 mg/L as CaCO3
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 16 h light: 8 h darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): immobilisation

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.68 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 0.53-0.88

Details on results:

- Mortality of control: blank control 0% (0/10), solvent control 10% (1/10) after 48 h

Reported statistics and error estimates:

LC50 values and their 95% confidence intervals were calculated by the moving average method of Thompson when the data permitted (constant dilution factor between the toxicant concentration). Otherwise, the LC50 was estimated by the method of Litchfield and Wilcoxon which requires at least two concentrations producing partial mortalities. If this method could not be used, the LC50 was estimated by graphical interpolation.

Cumulative mortalities:

0.08 mg/L:       0% after 24 and 48 hours

0.13 mg/L:       0% after 24 and 48 hours

0.22 mg/L:       0% after 24 and 48 hours

0.36 mg/L:       0% after 24 and 48 hours

0.60 mg/L:       30% after 24 hours, 40% after 48 hours

1.00 mg/L:       70% after 24 hours, 80% after 48 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to Daphnia magna in a bioassay screening the acute toxicity (48h) of the test substance.

Description of key information

EC50(48h) = 0.3 mg/L (nominal, vehicle) for Daphnia magna (US-EPA method)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.3 mg/L

Additional information

Several studies investigating the short-term toxicity of N-1-naphthylanilin to aquatic invertebrates are available. In the key study, the toxicity of the test substance to Daphnia magna was tested in a static system with soft water following US EPA recommendations (Sikka et al. 1981). Nominal concentrations of 0, 0.13, 0.22, 0.36, 0.60 and 1.00 mg/L were tested, with ethanol as solvent. This study yielded an EC50 value for immobilisation (48h) of 0.3 mg/L. Similar results were obtained in supporting studies, conducted during the same comprehensive investigation under varying conditions (Sikka et al. 1981). For adult Daphnia, an EC50 value (48h) of 0.68 mg/L was obtained in a static test system with soft water, and for first-instar Daphnia, additional EC50 values (48h) of 0.68 mg/L and 0.67 mg/L resulted from exposure in a semi-static test system witht hard water and soft water, respectively. During these two studies, nominal concentrations of 0, 0.08, 0.13, 0.22, 0.36, 0.60 and 1.0 mg/L were tested. Another study performed by the South Korean Ministry of Environment resulted in a comparable 48h-EC50 value for Daphnia magna of 0.58 mg/L. In order to be sufficently conservative, the lowest available 48h-EC50 value was used for the chemical safety assessment of the substance. The study obtained in the J-CHECK database of the Japanese Ministry of Environment was not considered for the hazard assessment, since man important information for a proper Klimisch rating is missing.