4-fluoroaniline

Endpoint summary

Description of key information

AOT: In Okazaki et al. the TDLo was determined to be 600 mg/kg bw.
ADT: Read –across approach to structural analogue substance 3,5-difluoroaniline (CAS 372-39-4) was used. The LD50 acute dermal toxicity in rat was determined to be  > 2000 mg/kg bw. 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Neither guideline nor GLP compliant study but nevertheless well documented and scientifically acceptable.

Qualifier:

no guideline available

Principles of method if other than guideline:

The test method is according to a neurotoxicity test. Based on a preliminary study a TDL0 of 600 mg/kg bw was determined which was used as application dose.

GLP compliance:

not specified

Test type:

fixed dose procedure

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 4 weeks
- Housing: Polycarbonate cages (three rats per cage)
- Diet: standard laboratory diet (MF, Oriental Yeast Co., Ltd, Tokyo, Japan)
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20-24 °C
- Humidity: 40-70%
- Photoperiod: 12-h light/dark

Route of administration:

oral: unspecified

Vehicle:

olive oil

Doses:

600 mg/kg bw

No. of animals per sex per dose:

9 (males)

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: All rats were carefully observed for mortality and clinical signs at 30 min and 5 h after dosing and once a day thereafter for the following 15 days. Body weights were recorded before dosing and on days 2, 4, 8, 15 and 16.
- Necropsy of survivors performed: Yes
- Other examinations performed:

- clinical signs:
Detailed clinical observations with functional tests were conducted on all rats 2 days before dosing and on day 9, and on all the surviving rats on day 15.
The observer first recorded each rat’s posture in the home cage, along with palpebral closure and the presence or absence of circling, writhing, biting, convulsions or vocalizations. Then the observer removed the rat, rating the ease of removal and handling, and recorded such signs as exophthalmos, crustiness around the eyes and piloerection. Extensor thrust, palpebral closure, fur appearance, lacrimation and salivation also were rated. The rat was placed on a field covered with a clean polypad. The rat was observed for 2 min and gait characteristics, arousal level and the number of supported or unsupported rearing and grooming episodes were recorded. At the end of 2 min, the number of faecal boluses and pools of urine on the field were recorded. Then the following reflex tests were carried out and recorded: approach response, touch response, finger-snap response, tail-pinch response, surface righting reflex and aerial righting reflex. Grip strength for fore- and hindlimbs was measured by a grip strength apparatus (Muromachi Kikai Co., Tokyo, Japan) and the mean value of two trials for each rat was recorded. Finally, the pads of the hind feet were painted with Indian ink and the rat was held in a horizontal position at a height of ca. 30 cm before being dropped onto paper. The distance between ink blots was measured and the mean value of two trials for each rat was recorded.

- histopathology:
Three rats from each experimental group were killed in numerical order on day 10 and the other rats were killed on day 16, under deep anaesthesia by an intraperitoneal injection of sodium pentobarbital. After cardiac perfusionfixation with a phosphate buffer solution containing paraformaldehyde (4%), glutaraldehyde (1%) and heparin (1000 IU/ L), the following organs and tissues were collected and fixed in 10% neutral buffered formalin solution: forebrain, including cerebral cortex, basal ganglia and corpus callosum; midbrain, including hippocampus, thalamus and hypothalamus; mesencephalon, including substantia nigria, colliculi, tectum and tegmentum; hindbrain, including cerebellum, pons and medulla oblongata; spinal cords, including cervical, thoracic and lumbar levels; trigeminal nerves with gasserian ganglia; and sciatic nerves. Tissues were processed for paraffin embedding, sectioned 5 μm thick, stained with haematoxylin and eosin (H&E) and examined by light microscopy.

Statistics:

The following data in all groups were analysed using multiple comparison tests: body weight, the number of faecal boluses and pools of urine on the open field, the number of grooming episodes and the number of supported or unsupported rearing events in the open field, data from the landing foot spray (cm) and the grip strength (g) for fore- and hindlimbs. They were first analysed by Bartlett’s test. If the group variance was determined to be homogeneous, all groups were compared by a one-way analysis of variance. If Bartlett’s test indicated heterogeneous variance, the Kruskal–Wallis test was employed and Dunnett’s test was used when there was a significant difference between the groups.
Results were expressed as the mean +/- standard deviation (SD). The other categorical and rank data obtained from the detailed clinical observations were analysed by the a × b chi-squared test, and when there was a significant difference Armitage’s chi-squared test was used to compare the difference between each halogenated aniline-treated group and the control group; P < 0.05 or P < 0.01 were considered significant for the statistical tests.

Sex:

male

Dose descriptor:

LDLo

Effect level:

600 mg/kg bw

Based on:

test mat.

Mortality:

None

Clinical signs:

other: - cyanosis between days 1 and 4 - only one rat showed decreased movement on day 1 - reduced response to extensor thrust of the hindlimb was found in on days 9 and 15 - gait abnormalities, including ataxia or hindlimb paralysis, were also found on days 9 a

Other findings:

Histopathology:
- A spongy change in the white matter of the spinal cords was found
- The lesions arose symmetrically and they were prominent in the lateral and ventral funiculi of the white matter. The lesions in the thoracic level were most prominent throughout the spinal cord. A spongy change in the tegmentum of the mesencephalon or the spinocellebelar tracts of the pons and medulla oblongata, and nerve fibre degeneration in the spinal nerves, trigeminal nerves, or sciatic nerves were also found.

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

600 mg/kg bw

Quality of whole database:

Neither guideline nor GLP compliant study but nevertheless well documented and scientifically acceptable.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

1994-02-21 to 1994-03-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

Adopted: 24 February 1987.

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Olac Ltd., Bicester, Oxon, England
- Age at study initiation: 7-10 weeks
- Weight at study initiation: 218 - 260 g
- Housing: Individually in metal cages with wire mesh floors
- Diet: Standard laboratory rodent diet (Biosure LAD 1), ad libitum
- Water: Drinking water, ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 - 23°C
- Humidity: 43%
- Air changes: 10 - 15 air changes per hour
- Photoperiod: 12 hours of artificial light in each 24 hours period

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: Dorso-lumbar region
- % coverage: 10
- Type of wrap if used: Gauze

REMOVAL OF TEST SUBSTANCE
- Washing: Yes, with warm water (30°C to 40°C)
- Time after start of exposure: At the end of the 24 hours period

TEST MATERIAL
- Amount applied: 2.33 mL/kg bw
- Concentration: 85.71%
- Constant volume or concentration used: yes

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Cages of rats were checked at least twice daily for any mortalities. Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1 (a period of five hours). On subsequent days animals were observed once in the morning and again at the end of the experimental day.
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs, body weight, organ weights, histopathology

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No

Clinical signs:

other: No signs of systemic reaction to treatment was observed. However, in two males, signs of reaction were apparent and comprised of piloerection, abnormal body carriage (hunched posture), abnormal gait (waddling), lethargy, decreased respiratory rate, pallor

Gross pathology:

No macroscopic abnormalities were observed for animals killed on Day 15.

Interpretation of results:

not classified

Remarks:

Migrated information

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline compliant study. The test substance was a structural analogue substance, please refer to IUCLID section 13 for read across justification.

Additional information

Acute oral toxicity

 

Key study

Okazaki et al., 2003

 

In the neither guideline nor GLP complaint literature source Okazaki et al. a neurotoxicity study after a single oral dose was conducted. The concentration of 600 mg/kg bw was applied orally to 9 male rats.

During the observation period of 15 days following clinical signs were observed:

- cyanosis between days 1 and 4

- only one rat showed decreased movement on day 1

- reduced response to extensor thrust of the hindlimb was found in on days 9 and 15

- gait abnormalities, including ataxia or hindlimb paralysis, were also found on days 9 and 15

- a significant decrease in grip strength of the hindlimbs was found in the on day 9 compared with the corresponding control group

- abnormal landing, such as on the animal’s side in the aerial righting reflex, was found on day 9

As no mortality was observed the TDLo was determined to be 600 mg/kg bw.

 

 

Supporting studies

 

ECHA disseminated dossier

 

As supporting studies following secondary sources can be mentioned which were found in the ECHA disseminated dossier (CAS 371-40-4).

In Dictionary of Substances and their effects, 2011, the LD 50 oral with rat was determined to be 420 mg/kg bw.

In RTECS, 2012, the LD 50 oral with rat was determined to be 417 mg/kg bw.

In HSDB, 1972, the LD 50 oral with rat was determined to be 460 mg/kg bw.

 

BASF SE, study no.: 78/176, 1980 (rat)

 

Read-across approach to structural analogue substance 3-Chloro-4-fluoroaniline (CAS 367-21-5) was used.

In a neither guideline nor GLP compliant acute oral toxicity study with male and female rats the test item was suspended in carboxymethyl cellulose and applied a t the concentration of 1000, 681, 464, 316 and 215 mg/kg bw to 5 animals per sex.

The animals were observed daily (body weight, clinical signs) over a period of 14 days.

Disponea, apathy, stagger, atony, clonic cramp, exsiccation cyanosis etc. were observed at different concentrations. After the observation period a necropsy was conducted with the dead and survived animals:

Following gross pathology effects were observed at the dead animals:

Hart: acute dilatation (right side), acute congestion hyperemia

Liver: lobule periphery; kidney: significant bright (nephrosis)

Stomach: bloody ulceration, grey brown colour

Musculature: multiple grey brown coloured

Intestine: hematis content

Urinary bladder: scattered noticeable contents

Further observations were not reported.

In this acute oral toxicity study the LD50 value was determined to be 480 mg/kg bw.

 

 

Acute dermal toxicity

 

Key study

Sandoz Crop Protection Ltd., study no. SNC 179/940230/AC, 1994

 

No data for this endpoint with the test substance is available. Thus, data from the similar substance 3,5-difluoroaniline (CAS 372-39-4) was summarised to cover this endpoint. For further justification see IUCLID section 13.

 

A study was performed to assess the acute dermal toxicity of 3,5-difluoroaniline to the rat. The method followed was that described in the OECD Guideline for Testing of Chemicals No. 402 “Acute Dermal Toxicity”. Adopted: 24 February 1987.

A group of six rats (three males and three females) was given a single dermal application of the test substance, at a maximum practical concentration of 85.71 % and at a dose level of 2.0 g/kg bodyweight. All animals were killed and examined macroscopically on Day 15, the end of the observation period.

There were no deaths and in the majority of animals no signs of systemic reaction to treatment.

However, in two males, signs of reaction to treatment were apparent and comprised of piloerection, abnormal body carriage (hunched posture), abnormal gait (waddling), lethargy, decreased respiratory rate, pallor of the extremities, Unsteadiness and increased lacrimation (red). Recovery in both rats, as judged by external appearance and behaviour, was complete by Day 6.

Sites of application of 3,5-difluoroaniline showed no irritation or other dermal changes.

Slightly low bodyweight gains were recorded for all three males an Day 8 and in two males and one female Day 15. All other rats achieved anticipated bodyweight gains throughout the study.

No abnormalities were recorded at the macroscopic examination on Day 15.

The acute lethal dermal dose to rats of 3,5-difluoroaniline was found to be greater than 2000 mg/kg bw.

 

 

Acute intraperitoneal toxicity

BASF SE, study no.: 78/176, 1980

 

Read-across approach to structural analogue substance 3-Chloro-4-fluoroaniline (CAS 367-21-5) was used.

In this study with female and male mice the test item was applied intaperitoneal.

The animals were observed daily (body weight, clinical signs) over a period of 14 days. After the observation period a necropsy was conducted with the dead and survived animals:

Clinical signs of dyspnoea, apathy, stagger, tremble, spastic gait, balance disturbance, clonic cramp, cyanosis etc were observed at the tested concentration of 700 mg/kg bw.

At the concentration of 200 mg/kg bw only dyspnoea, agitation, stagger and cyanosis were observed.

At the gross pathology no intra-abdominal substance precipitation or adhesion was observed

The LD50 in female and male mice was determined to be 700 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
Neither guideline nor GLP compliant study but nevertheless well documented and scientifically acceptable.

Justification for selection of acute toxicity – dermal endpoint
GLP and guideline compliant study. The test substance was a structural analogue substance, please refer to IUCLID section 13 for read across justification.

Justification for classification or non-classification

Acute oral toxicity

 

Dangerous Substance Directive (67/548/EEC)
The available study is considered reliable and suitable for classification purposes under Directive 67/548/EEC. As a result the substance is considered to be classified for acute oral toxicity Xn; R22: Harmful if swallowed under Directive 67/548/EEC, as amended for the 31st time in Directive 2009/2/EG.

 

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is considered to be classified for acute toxicity cat. 4, H302: Harmful if swallowed under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation (EC) No 605/2014.

 

 

Acute dermal toxicity

 

Dangerous Substance Directive (67/548/EEC)
The available study is considered reliable and suitable for classification purposes under Directive 67/548/EEC. As a result the substance is not considered to be classified for acute dermal toxicity under Directive 67/548/EEC, as amended for the 31st time in Directive 2009/2/EG.

 

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is considered not to be classified for acute dermal toxicity under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation (EC) No 605/2014.