3-aminophenol

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

adult fish: sub(lethal) effects

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From December 29, 1995 to March 28, 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed according to the guideline.

Qualifier:

according to guideline

Guideline:

OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)

Deviations:

no

Principles of method if other than guideline:

Long term toxicity study was conducted on fish according to the OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study).

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations and sampling method: test solutions were sampled from the middle layers of 2 vessels of each test group at the beginning of exposure (day 0), day 3, day 7, day 14 and day 21, and mixed: thereafter, they were analyzed by HPLC as it is or after dilution.
- Sample storage conditions before analysis: no data

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:Test substance was added to dilution water, dissolved by ultrasonication procedure, and the test substance solution of 10,000 mg/L was prepared. This test substance solution was diluted when necessary to prepare stock solutions of concentrations 10 times higher each test concentration group (100 mg/L group: 1,000 mg/L, 50 mg/L group: 500 mg/L, 25.0 mg/L group: 250 mg/L, 12.5 mg/L group: 125 mg/L, 6.25 mg/L group: 62.5 mg/L).
Test solution was prepared by mixing the stock solution and dilution water in a constant flow rate (stock solution: 2.5 mL/min. dilution water: 22.5 mL/min.) The mixed test solutions were divided in two using splitter cells and sent to the individual testing containers.

- Eluate: /
- Differential loading: /
- Controls: dilution water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): n.a.
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): n.a.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No; At the beginning of exposure, test solutions of the concentration groups of 50 mg/L or higher was slight yellow and transparent, and that of the lower concentration groups was colorless and transparent. These properties did not change until the end of exposure.

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Orange killifish
- Strain: /
- Source: Nakajima fish farm (Daimyojin Nagasu-machi Tamana-gun, Kumamoto 869-01), Lot No. LNR-951216; Purchased date: November 28, 1995
- Age at study initiation (mean and range, SD): /
- Length at study initiation (length definition, mean, range and SD): 1.83 cm (1.61 – 2.03cm), n=120
- Weight at study initiation (mean and range, SD): 0.0925 g (0.0669 – 0.127g), n=120
- Method of breeding: /
- Feeding during test: Yes
- Food type: commercially available TetraMIn
- Amount: 2% of fish body weight; In the case number of individuals declined by death, feeding amount was decreased accordingly.
- Frequency: every day

ACCLIMATION
- Acclimation period: For acclimation, the fish were raised for 12 or more days
- Acclimation conditions (same as test or not): under the same conditions (water quality, temperature, etc.) as the test conditions. At the start of acclimation there was a 24-hour drug bath in still water using Elverge (Ueno fine chemicals industry)
- Type and amount of food: Commercially available TetraMIn (Tetra Werke) was given as food.
- Feeding frequency: /
- Health during acclimation (any mortality observed): Healthy and macroscopically normal individuals were used for the test. Mortality rate during the 7 days before the beginning of exposure was 5% or less. Besides, 96-hour LC50 of reference substance (Copper(II) sulfate pentahydrate, special grade chemical) was 0.919 mg/L.

QUARANTINE (wild caught) n.a.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

35.5 mg/L as CaCO3

Test temperature:

24±1°C (23.1-24.9°C)

pH:

7.27-7.74

Dissolved oxygen:

7.4-8.3 mg/L

Nominal and measured concentrations:

Nominal concentrations: 100, 50, 25.0, 12.5, 6.25 mg/L
Measured concentrations: Measured values of test substance concentration during exposure period were 5.88 – 105 mg/L (nominal value 6.25 – 100 mg/L), which were in the range of 89.7 – 105% of the nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel, Material, size, headspace, fill volume: 3.0 L glass container (16cm in diameter, 17cm in depth). Water discharge holes were placed in the sides of the containers in order to maintain a volume of approximately 1.8 L. Pierced a hole for drainage on the side surface of the container so that the internal volume becomes about 1.8 L.
- Type (delete if not applicable): open
- Aeration: none
- Type of flow-through (e.g. peristaltic or proportional diluter): A diluter which continuously supplies test water diluted to the required concentration levels from stock solution was used. Metering pump: Glass microscale metering pump GMW-A type (TOKYO RIKAKIKAI CO., LTD.)
- Renewal rate of test solution (frequency/flow rate): flow rate of 12,5 mL/min. per test vessel. This flow rate is equivalent to the renewal rate of test water about 10 times/day.
- No. of organisms per vessel: 10 fishes per vessel
- No. of vessels per concentration (replicates): 2 vessels per test group/concentration/5 concentrations
- No. of vessels per control (replicates): 2 vessels
- Biomass loading rate: 0.51 g/l

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated tap water (removed residual chlorine, etc from tap water supplied by Kurume city by processing with activated carbon, and it was sufficiently aerated)
- Total organic carbon: COD: 0.2 mg/L
- Metals: not detected as for Hg, Cu, Cd, Pb, Al, Ni, Cr, Sn. Zn: 0.01, Fe: 0.02 mg/L (see annexe II of the report for details)
- Pesticides: not detected (see annexe II of the report for details)
- Chlorine: free chlorine not detected
- Alkalinity: 21.0 mg/L
- Ca/mg ratio: total hardness: 35.5 mg/L as CaCO3 (Ca: 9.2, Mg: 3.06 mg/L
- Conductivity:147 µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: each three or four days

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8
- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, growth rate, behaviour

TEST CONCENTRATIONS
- Spacing factor for test concentrations 2
- Justification for using less concentrations than requested by guideline: n.a.
- Range finding study
- Test concentrations and Results used to determine the conditions for the definitive study: Based on the 96-h LC50 value (121 mg/L) obtained from fish acute toxicity test, observed symptoms and the properties of test substance

Reference substance (positive control):

yes

Remarks:

Copper(II) sulfate pentahydrate

Key result

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: Nontoxic

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: toxic symptoms and food consumption

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

- Mortality/survival at embryo and larval stages: n.a.
- Overall mortality/survival: The lowest tested concentration (Lowest Lethal Concentration) of the concentration groups at which dead individuals were observed was 100 mg/L (nominal concentration).
The lowest tested concentration (Lowest Observed Effect Concentration) of the concentration groups at which effects other than death were observed in test fishes was 50 mg/L (nominal concentration).
Any of Median Lethal Concentrations (LC50) based on nominal concentrations of day 7, day 14, and day 21 were > 100 mg/L.
- Days to hatch and numbers hatched:n.a.
- Data for length and weight of surviving fish: With regard to the growth rate based on the body weight of Orange killifish exposed to m-Aminophenol for 21 days, no significant difference was demonstrated at the concentration groups of 50 mg/L or less (nominal concentrations) compared with control group.
(p=0.05, statistical method, one-way analysis of variance)
- Type of and number with morphological abnormalities and with behavioural abnormalities: As toxicity symptoms, declined activity and aggregation at surface layer were observed in the 100 mg/L group. No abnormal symptom was observed during exposure period in the control group.
Though the decline of food intake activity of Orange killifish was found at early time of the exposure period in the concentration groups of 100 mg/L and 50 mg/L, it was recovered thereafter. No decline of food intake was found in the other concentration groups.
- Other biological observations: /
- Effect concentrations exceeding solubility of substance in test medium: no
- Incidents in the course of the test which might have influenced the results: no

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: 96-hour LC50 of reference substance (Copper(II) sulfate pentahydrate, special grade chemical) was 0.919 mg/L

Reported statistics and error estimates:

2) Test for significant difference in growth rate
Regarding the calculated growth rates, the presence or absence of significant differences compared with control group was tested by One-way analysis of variance. However, 100 mg/L group at which death was observed more than 10% at the end of exposure was excluded from the test.
NOEC was assessed by taking into account of the results of this test for significant difference in growth rate as well as the mortality or observed symptoms, etc. of test organisms.
(p=0.05, statistical method, one-way analysis of variance)

Validity criteria fulfilled:

yes

Remarks:

no mortality in the control, O2 >= 60% saturation

Conclusions:

Non-Observed Effect Concentration for 21 days (NOEC) = 25.0 mg/L (based on nominal values).
The highest concentration of the concentration groups at which no dead individual was observed was 50 mg/L, the highest concentration at which no toxic symptom was observed was 25.0 mg/L, the highest concentration at which no abnormality in food consumption status was found was 25.0 mg/L, and the highest concentration at which no significant difference was suggested compared with control group in the growth rate of test organisms was 50 mg/L.

Executive summary:

The test item was tested according to OECD 204 guideline, prolonged toxicity test in fish, and GLP. The test was conducted in flow-through conditions on the orange killifish (Oryzias latipes) during 21 days. The tested concentrations were 6.25, 12.5, 25.0, 50.0 and 100 mg/L. Analytical monitoring showed that measured concentrations were in the range of 89.7% to 105% of the nominal concentrations. Mortality, growth rate, other toxic symptoms and behaviour were evaluated. Non-Observed Effect Concentration for 21 days (NOEC) = 25.0 mg/L (based on nominal values). The highest concentration of the concentration groups at which no dead individual was observed was 50 mg/L, the highest concentration at which no toxic symptom was observed was 25.0 mg/L, the highest concentration at which no abnormality in food consumption status was found was 25.0 mg/L, and the highest concentration at which no significant difference was suggested compared with control group in the growth rate of test organisms was 50 mg/L. LC50 at day 7, 14, and 21 were all > 100 mg/L (nominal concentrations). All the validity criteria were met.

Description of key information

The test item was tested according to OECD 204 guideline, prolonged toxicity test in fish, and GLP. The test was conducted in flow-through conditions on the orange killifish (Oryzias latipes) during 21 days. The tested concentrations were 6.25, 12.5, 25.0, 50.0 and 100 mg/L. Analytical monitoring showed that measured concentrations were in the range of 89.7% to 105% of the nominal concentrations. Mortality, growth rate, other toxic symptoms and behaviour were evaluated. Non-Observed Effect Concentration for 21 days (NOEC) = 25.0 mg/L (based on nominal values). The highest concentration of the concentration groups at which no dead individual was observed was 50 mg/L, the highest concentration at which no toxic symptom was observed was 25.0 mg/L, the highest concentration at which no abnormality in food consumption status was found was 25.0 mg/L, and the highest concentration at which no significant difference was suggested compared with control group in the growth rate of test organisms was 50 mg/L. LC50 at day 7, 14, and 21 were all > 100 mg/L (nominal concentrations). All the validity criteria were met.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

25 mg/L

Additional information

The test item was tested according to OECD 204 guideline, prolonged toxicity test in fish, and GLP. The test was conducted in flow-through conditions on the orange killifish (Oryzias latipes) during 21 days. The tested concentrations were 6.25, 12.5, 25.0, 50.0 and 100 mg/L. Analytical monitoring showed that measured concentrations were in the range of 89.7% to 105% of the nominal concentrations. Mortality, growth rate, other toxic symptoms and behaviour were evaluated. Non-Observed Effect Concentration for 21 days (NOEC) = 25.0 mg/L (based on nominal values). The highest concentration of the concentration groups at which no dead individual was observed was 50 mg/L, the highest concentration at which no toxic symptom was observed was 25.0 mg/L, the highest concentration at which no abnormality in food consumption status was found was 25.0 mg/L, and the highest concentration at which no significant difference was suggested compared with control group in the growth rate of test organisms was 50 mg/L. LC50 at day 7, 14, and 21 were all > 100 mg/L (nominal concentrations). All the validity criteria were met.