4-fluoroaniline

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2010-11-26 to 2011-02-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to generally valid and/or internationally accepted testing guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-fluoroaniline

EC Number:

206-735-5

EC Name:

4-fluoroaniline

Cas Number:

371-40-4

Molecular formula:

C6H6FN

IUPAC Name:

4-fluoroaniline

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The concentrations of the test item were analysed in the duplicate test media sampIes from the four highest test concentrations from all sampling times (0, 24 and 96 hours). From the control sampIes only one of the duplicate sampIes was analysed from each of all sampling times.
- Sampling method: One sample of the stock solution and duplicate samples from the freshly prepared test media of all test concentrations and the control were taken at the start of the test. For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of all test concentrations and the control were collected at the end of the test (after 96 hours) by pouring together the contents of the test beakers of each treatment. However, from the test concentrations of 45 and 100 mg test item/L duplicate sampIes were taken after 1 day, respectively, because all fish died.
- Storage of sampIes: All sampIes were stored in a freezer (=< - 10°C), protected from light until analysis was performed.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTIONS.

- Method: A stock solution of nominal 500 mg/L was prepared by dissolving 2101.2 mg test item into 4202.4 mL test water by intense stirring for 40 min and ultrasonic treatment for 2 x 5 minutes. Then, adequate volumes of this stock solution were mixed into test water to prepare the test media of the desired test concentrations.
- Control: Reconstituted water according to guideline. In the control, test water was used without addition of the test item.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow Trout (Oncorhynchus mykiss)
- Source: The test fish were obtained from Forellenzuchtbetrieb Stärk, 88348 Bad Saulgau, Germany
- Age at study initiation: Juveniles
- Length at study initiation: The mean body length of the fish in the test was 5.08 cm ± 0.38 cm (Mean ± SD).
- Weight at study initiation: The mean body wet weight 1.26 ±0.34 g (Mean ± SD).
- Sex: Male and female
- Feeding during the test: None

ACCLIMATION
- Acclimation period: All fish were obtained and held in the laboratory for at least 12 days before the start of the test.
- Acclimation conditions: They were held in water of the quality to be used in the test for at least seven days immediately before testing under the following conditions:
Light: 12 - 16 hours photoperiod daily
Temperature: 13 - 17 °C
Oxygen concentration: at least 80 % of the air saturation value
- Feeding frequency: Three times per week or daily until 24 hours before the test was started
- Health during acclimation: During the last 7 days prior to the start of the test 8 fish (7 %) died in the test fish batch. Nevertheless, the fish batch was accepted since mortality was < 1 % during the first month of holding. Furthermore, no mortality at aII occurred in the control and the two lowest concentrations of the test and a well defined dose-response was determined for the three higher test concentrations.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

2.5 mmol/L (= 250.0 mg/L) as CaCO3

Test temperature:

13 °C - 15 °C

pH:

7.6 to 8.0

Dissolved oxygen:

92-100%;

Nominal and measured concentrations:

- Nominal concentrations: 100, 45, 21, 9.4 and 4.3 mg test item/L and a control.
- Measured concentrations: At the start of the test just before introduction of the fish 80 % of the nominal test concentrations were measured. After 96 hours test duration 82% of the nominal values were determined. Thus, during the test period of 96 hours the fish were exposed to a mean of 81 % of nominal. Therefore, all reported results are related to nominal concentrations of the test item. The mean recovery in the test samples was determined to be 80 % in the freshly prepared samples and 82 % in the aged samples.

Details on test conditions:

TEST SYSTEM
- Test vessel: 12 litre glass aquaria with 10 litre test medium
- Aeration: The test media were slightly aerated during the test
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water according to guideline.
- Intervals of water quality measurement: The water temperature, pH-values and the dissolved oxygen concentrations were determined daily in the test media of each treatment group.

OTHER TEST CONDITIONS
- pH: 7.6 to 8.0
- Adjustment of pH: No
- Photoperiod: 16 h light: 8 h dark
- Light intensity: 480-1060 lux

EFFECT PARAMETER MEASURED: Mortality and sublethal effect.

TEST CONCENTRATIONS
- Nominal concentrations: 100, 45, 21, 9.4 and 4.3 mg test item/L and a control.
- Measured concentrations: At the start of the test just before introduction of the fish 80% of the nominal test concentrations were measured. After 96 hours test duration 82 % of the nominal values were determined. Thus, during the test period of 96 hours the fish were exposed to a mean of 81 % of nominal. Therefore, all reported results are related to nominal concentrations of the test item.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

22.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: n.d.

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

45 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% conf. limits (49.8 - 44.36)

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

21 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: n.d.

Effect concentrations 4

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

9.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: n.d.

Details on results:

Based on the test results the 96-hour LC50 of the test item for Rainbow Trout (Oncorhynchus mykiss) was determined to be nominal 22.7 mg test item/L. The no observed effect concentration (NOEC) was determined to be 9.4 mg test item/L, the LOEC was determined to be 21 mg test item/L and the LC50 was determined to be 22.7 mg/L. All values are based on nominal test concentrations. All validity criteria were met.

Reported statistics and error estimates:

The LC50 at the observation times was calculated by Probit analysis.
The NOEC, the LOEC, the LC0 and the LC100 were determined directly from the raw data.
The software used to perform the statistical analysis was ToxRat Professional, Version 2.10.05, ToxRat® Solutions GmbH.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In the static short-term toxicity to fish test according to the OECD guideline No. 203 (1992), the 96-hour LC50 of the test substance for Rainbow Trout (Oncorhynchus mykiss) was determined to be 22.7 mg test item/L nominal. The no observed effect concentration (NOEC) was determined to be 9.4 mg test item/L, the LOEC was determined to be 21 mg test item/L and the LC50 was determined to be 22.7 mg/L, all values also based on nominal test concentration. All validity criteria were met.

Executive summary:

The acute toxicity to fish Rainbow Trout (Oncorhynchus mykiss) of the test substance was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation No. 440/2008/ C.1 (2008). This study encompassed 6 treatment groups (5 dose rates of the test item, control) each containing 7 individuals. The acute toxicity to unfed juvenile Rainbow Trout was determined in an aerated, static, 96-hour test. The test fish were observed after approximately 2, 24, 48, 72 and 96 hours test duration for sublethaI effects and mortality. Dead fish were removed at least once daily and discarded. The sampIes of the test medium were analysed via HPLC-method at start and after 96 hours. The test concentrations were: 100, 45, 21, 9.4 and 4.3 mg test item/L and a control. Water quality parameters pH-value, temperature and oxygen-saturation, measured after 0, 24, 48, 72 and 96 h were determined to be within the acceptable Iimits. The no observed effect concentration (NOEC) was determined to be 9.4 mg test item/L, the LOEC was determined to be 21 mg test item/L and the LC50 was determined to be 22.7 mg/L. All values also based on nominal test concentrations.