N-1-naphthylaniline

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

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

Principles of method if other than guideline:

96h acute toxicity test

GLP compliance:

not specified

Vehicle:

not specified

Test organisms (species):

not specified

Test type:

not specified

Water media type:

not specified

Total exposure duration:

96 h

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.7 mg/L

Validity criteria fulfilled:

not specified

Reference 2

Endpoint:

short-term toxicity to fish

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:

96h acute bioassay using rainbow trout 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: purified PNA (as hydrochloride salt) dissolved in ethanol was added to well-aerated test water, test solutions were vigorously stirred with a glass rod
- Controls: yes, vehicle control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not exceeding 0.5 mL/L water (recommended by US EPA)

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: commercial fish hatcheries
- Weight at study initiation (mean and range, SD): 0.6 g
- Feeding during test: no

ACCLIMATION
- Acclimation period: no data
- Acclimation conditions (same as test or not): maintained in round fiberglass tanks, flow-through at 13+/-2 °C, 16 h light: 8 h dark
- Type and amount of food: commercial trout feed
- Health during acclimation (any mortality observed): less than 5% mortality observed for more than one year

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

118 mg/L as CaCO3

Test temperature:

13 +/-2 °C

pH:

not specified

Dissolved oxygen:

not specified

Nominal and measured concentrations:

Nominal: 0.13, 0.22, 0.36, 0.60 and 1 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 5 gallon wide-mouth glass jars, containing 15 L of tap water
- Aeration: no data
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 0
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.4 g fish/L test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Syracuse municipal water filtered and dechlorinated through active charcoal
- Total organic carbon: 2.2 +/- 1.7 mg/L
- Particulate matter: < 5 mg/L
- Chlorine: < 10 µg/L
- Alkalinity: 117.9 +/- 10.4 mg/L
- Conductivity: 265+/- 17.3 µmhos/cmL
- Culture medium different from test medium: no

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, recorded daily

TEST CONCENTRATIONS
- 0.13, 0.22, 0.36, 0.60 and 1 mg/L
- Spacing factor for test concentrations: at least 0.6
- Range finding study
- Test concentrations: a series of concentrations (dilution factor of at least 0.6)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.44 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%

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.

Sublethal observations / clinical signs:

Cumulative mortalities:

0.13 mg/L:       0% after 24, 48 and 96 hours

0.22 mg/L:       0% after 24, 48 and 96 hours

0.36 mg/L:       0% after 24 and 48 hours, 10% after 96 hours

0.60 mg/L:       0% after 24 hours, 100% after 48 and 96 hours

1.00 mg/L:       20% after 24 hours, 70% after 48 hours and 100% after 96 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to rainbow trout in a bioassay screening the acute toxicity (96h) of the test substance.

Reference 3

Endpoint:

short-term toxicity to fish

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:

192h acute bioassay using bluegill sunfish as test organism (following recommendations of US-EPA)

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

- Sampling method: samples were extracted in a separatory funnel with 20 mL methylene chloride; measured daily

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution of PNA dissolved in acetone
- Controls: yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not exceeding 0.5 mL/L water (recommended by US EPA)

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill sunfish
- Source: commercial fish hatcheries
- Weight at study initiation (mean and range, SD): 1.8 g
- Feeding during test: yes, every other day

ACCLIMATION
- Acclimation period: no data
- Acclimation conditions (same as test or not): maintained in round fiberglass tanks, flow-through at 20+/-2 °C, 16 h light: 8 h dark
- Type and amount of food: commercial trout feed
- Health during acclimation (any mortality observed): less than 5% mortality observed for more than one year

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

192 h

Hardness:

118 mg/L as CaCO3

Test temperature:

20 +/-2 °C

pH:

not specified

Dissolved oxygen:

not specified

Nominal and measured concentrations:

Nominal: 0.11, 0.18, 0.35 and 0.53 mg/L (Test 1)
Nominal: 0.24, 0.38, 0.64 and 1.05 mg/L (Test 2)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 30L aquaria
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): 3 L/hour (corresponding to 2.4 tank volumes/day, 90% replacement time of about one day)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 0
- Biomass loading rate: 0.6 g fish/L test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Syracuse municipal water filtered and dechlorinated through active charcoal
- Total organic carbon: 2.2 +/- 1.7 mg/L
- Particulate matter: < 5 mg/L
- Chlorine: < 10 µg/L
- Alkalinity: 117.9 +/- 10.4 mg/L
- Conductivity: 265+/- 17.3 µmhos/cmL
- Culture medium different from test medium: no

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, recorded daily

TEST CONCENTRATIONS
- 0.11, 0.18, 0.35 and 0.53 mg/L (nominal) in Test 1
0.24, 0.38, 0.64 and 1.05 mg/L (nominal) in Test 2
- Spacing factor for test concentrations: at least 0.6
- Range finding study
- Test concentrations: a series of concentrations (dilution factor of at least 0.6)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.82 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%

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.

Sublethal observations / clinical signs:

Cumulative mortalities:

TEST 1:

0.11 mg/L:       0% after 48, 96, 144 and 192 hours

0.18 mg/L:       0% after 48, 96, 144 and 192 hours

0.35 mg/L:       0% after 48 and 96 hours, 10% after 144 and 192 hours

0.53 mg/L:       0% after 48 and 96 hours, 60% after 144 hours and 90% after 192 hours

TEST 2:

0.24 mg/L:       0% after 48, 96, 144 and 192 hours

0.38 mg/L:       0% after 48, 96 and 144 hours, and 10% after 192 hours

0.64 mg/L:       0% after 48 hours, 10% after 96 hours, 90% after 144 hours and 100% after 192 hours

1.05 mg/L:       40% after 48 hours, 90% after 96 hours, and 100% after 144 and 192 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to bluegill sunfish in a bioassay screening the acute toxicity (192h) of the test substance.

Reference 4

Endpoint:

short-term toxicity to fish

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:

96h acute bioassay using bluegill sunfish 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: purified PNA (as hydrochloride salt) dissolved in ethanol was added to well-aerated test water, test solutions were vigorously stirred with a glass rod
- Controls: yes, vehicle control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not exceeding 0.5 mL/L water (recommended by US EPA)

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill sunfish
- Source: commercial fish hatcheries
- Weight at study initiation (mean and range, SD): 1.0 g
- Feeding during test: no

ACCLIMATION
- Acclimation period: no data
- Acclimation conditions (same as test or not): maintained in round fiberglass tanks, flow-through at 20+/-2 °C, 16 h light: 8 h dark
- Type and amount of food: commercial trout feed
- Health during acclimation (any mortality observed): less than 5% mortality observed for more than one year

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

192 h

Hardness:

118 mg/L as CaCO3

Test temperature:

20 +/-2 °C

pH:

not specified

Dissolved oxygen:

not specified

Nominal and measured concentrations:

Nominal: 0.13, 0.22, 0.36, 0.60 and 1 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 5 gallon wide-mouth glass jars, containing 15 L of tap water
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.7 g fish/L test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Syracuse municipal water filtered and dechlorinated through active charcoal
- Total organic carbon: 2.2 +/- 1.7 mg/L
- Particulate matter: < 5 mg/L
- Chlorine: < 10 µg/L
- Alkalinity: 117.9 +/- 10.4 mg/L
- Conductivity: 265+/- 17.3 µmhos/cmL
- Culture medium different from test medium: no

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, recorded daily

TEST CONCENTRATIONS
- 0.13, 0.22, 0.36, 0.60 and 1 mg/L
- Spacing factor for test concentrations: at least 0.6
- Range finding study
- Test concentrations: a series of concentrations (dilution factor of at least 0.6)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.82 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%

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.

Sublethal observations / clinical signs:

Cumulative mortalities:

0.13 mg/L:       0% after 48, 96, 144 and 192 hours

0.22 mg/L:       11% after 48 hours and 22% after 96, 144 and 192 hours

0.36 mg/L:       30% after 48, 96, 144 and 192 hours

0.60 mg/L:       0% after 48 hours, 10% after 96 hours, 40% after 144 hours and 80% after 192 hours

1.00 mg/L:     0% after 48 hours, 78% after 96 hours and 100% after 144 and 192 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to bluegill sunfish in a bioassay screening the acute toxicity (96h) of the test substance.

Reference 5

Endpoint:

short-term toxicity to fish

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:

192h acute bioassay using rainbow trout as test organism (following recommendations of US-EPA)

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

- Sampling method: samples were extracted in a separatory funnel with 20 mL methylene chloride; measured daily

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution of PNA dissolved in acetone
- Controls: yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not exceeding 0.5 mL/L water (recommended by US EPA)

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: commercial fish hatcheries
- Weight at study initiation (mean and range, SD): 4.4 g
- Feeding during test: yes, every other day

ACCLIMATION
- Acclimation period: no data
- Acclimation conditions (same as test or not): maintained in round fiberglass tanks, flow-through at 13+/-2°C, 16 h light: 8 h dark
- Type and amount of food: commercial trout feed
- Health during acclimation (any mortality observed): less than 5% mortality observed for more than one year

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

192 h

Post exposure observation period:

no data

Hardness:

118 mg/L as CaCO3

Test temperature:

13 +/-2 °C

pH:

no data

Dissolved oxygen:

no data

Salinity:

not relevant

Nominal and measured concentrations:

0.11, 0.24, 0.36 and 0.67 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 30L aquaria
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): 3 L/hour (corresponding to 2.4 tank volumes/day, 90% replacement time of about one day)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 0
- Biomass loading rate: 1.5 g fish/L test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Syracuse municipal water filtered and dechlorinated through active charcoal
- Total organic carbon: 2.2 +/- 1.7 mg/L
- Particulate matter: < 5 mg/L
- Chlorine: < 10 µg/L
- Alkalinity: 117.9 +/- 10.4 mg/L
- Conductivity: 265+/- 17.3 µmhos/cmL
- Culture medium different from test medium: no


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


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, recorded daily


TEST CONCENTRATIONS
- 0.11, 0.24, 0.36 and 0.67 mg/L (nominal)
- Spacing factor for test concentrations: at least 0.6
- Range finding study
- Test concentrations: a series of concentrations (dilution factor of at least 0.6)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.74 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%

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.

Sublethal observations / clinical signs:

Cumulative mortalities:

0.11 mg/L:       0% after 48, 96, 144 and 192 hours

0.24 mg/L:       0% after 48 hours and 10% after 96, 144 and 192 hours

0.36 mg/L:       0% after 48 hours, 10% after 96 hours, 70% after 144 hours and 90% after 192 hours

0.67 mg/L:       0% after 48 hours, 60% after 96 hours and 100% after 144 and 192 hours

Validity criteria fulfilled:

not applicable

Conclusions:

The test substance showed to be acutely very toxic to rainbow trout in a bioassay screening the acute toxicity (192h) of the test substance.

Description of key information

LC50(96h) = 0.44 mg/L (nominal, vehicle) for Oncorhynchus mykiss (US-EPA method)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.44 mg/L

Additional information

Several studies investigating the short-term toxicity of N-phenyl-1-naphthylamine to fish is availbale. In the key study, the toxicity of the test substance was tested in a semi-static system following US-EPA recommendations using Oncorhynchus mykiss as test organism (Sikka et al. 1981). Nominal concentrations of 0.13, 0.22, 0.36, 0.60 and 1 mg/L were tested, with ethanol as solvent. This study yielded an LC50 value (96h) of 0.44 mg/L. Similar results were obtained in supporting studies, conducted during the same comprehensive investigation with various fish species and test conditions (Sikka et al. 1981). For Lepomis macrochirus, an LC50 value (96h) of 0.82 mg/L was obtained both in a semi-static and flow-through test system, and for Oncorhynchus mykiss, an additional LC50 value (96h) of 0.74 mg/L resulted from exposure in a flow-through test system. In order to be sufficently conservative, the lowest available 96h-LC50 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.