2,4,6-trichloroaniline

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

The NOAEL and LOAEL was considered to be 0.3 mg/kg/day and 3 mg/kg/day for F0 generation respectively and LOAEL was considered to be  3 mg/kg/day  for F1 generation when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months.

Link to relevant study records

Reference

Endpoint:

one-generation reproductive toxicity

Remarks:

based on generations indicated in Effect levels (migrated information)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from secondary source .

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

A chronic reproductive toxicity study of 2, 4, 6-trichloroaniline iwas conducted in white male and female rats by oral gavage.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 2,4,6-Trichloroaniline
- Molecular formula (if other than submission substance): C6-H4-Cl3-N
- Molecular weight (if other than submission substance): 196.464
- Substance type: Organic
- Physical state: Solid

Species:

rat

Strain:

not specified

Details on species / strain selection:

White rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not specified

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

other: not applicable

Vehicle:

other: Oil solution

Remarks:

Not specified

Details on exposure:

Details on exposure
PREPARATION OF DOSING SOLUTIONS: 2,4,6-trichloroaniline were mix with oil solution in the concentration of at doses of 0.4, 4, or 40 mg/kg-day, 5 days/week (adjusted to 0.3, 3, or 29 mg/kg-day).

VEHICLE
- Justification for use and choice of vehicle (if other than water): Oil solution were used
- Concentration in vehicle: 0, 0.3, 3 and 29 mg/kg/day

Details on mating procedure:

The animals were mated at the end of the 6-month treatment period.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

6 months

Frequency of treatment:

5days /week

Details on study schedule:

No data available

Remarks:

Doses / Concentrations:
0, 0.3, 3.0, and 29 mg/kg/day
Basis:
no data

No. of animals per sex per dose:

120 males and 60 females

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Parental animals: Observations and examinations:

Observations and examinations performed & frequency
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked in table [No.?] were included: Mortality was observed.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: On every 30 days throughout the treatment period.

BODY WEIGHT: Yes
- Time schedule for examinations: On every 30 days throughout the treatment period.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: On every 30 days of treatment period.
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: All 180 animals were examined.
- Parameters checked in table [No.?] were examined: Methemoglobin, conditioned reflexes, Concentration of formed elements and serum hemoglobin, Polychromaphilic and hypochromic RBCs and WBC were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: On every 30 days of treatment period.
- Animals fasted: No data available
- How many animals: All 180 animals were examined.
- Parameters checked in table [No.?] were examined: : Residual nitrogen, pyruvic acid, catalase, alanine aminotransferase [ALT] and aspartate aminotransferase [AST] levels, nitrogen and pyruvic acid in serum and lactate dehydrogenase (LDH), succinic dehydrogenase (SDH) and Chromosomal aberrations activities in the liver and kidney were examined.

Oestrous cyclicity (parental animals):

Any irregularities in the estrous cycle ovogenesis were investigated.

Sperm parameters (parental animals):

Sperm parameters (parent animal)
Parameters examined in [all P] male parental generations : Spermatogenesis were observed

Litter observations:

Number of live embreyos, Number of dead embreyos and numbers of fetuses/dam were observed

Postmortem examinations (parental animals):

Postmortem examinations (Parent Animal)
SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: Yes

- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]: Number of implantation sites was examined.

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic and abdominal viscera.]: Yes, at the termination of study.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively: Yes

Organ weights were examined.

Organ weighted:
Brain, Liver, kidneys and reproductive organs were examined.

Postmortem examinations (offspring):

Postmortem examinations (offspring)
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age: No data available

- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: Yes

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]: Embryotoxicity and teratogenicity were examined.

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively. : Yes, morphofunctional indices were examined.

Statistics:

No data available

Reproductive indices:

Fertility index, gestation index and implantation index were examined.

Offspring viability indices:

No data available

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Description (incidence and severity):

Body weight gain: In 29 mg/kg/day treated rat decreased in weight gain were observed as compared to control.

Food consumption and compound intake (if feeding study):

not specified

Description (incidence and severity):

Body weight gain: In 29 mg/kg/day treated rat decreased in weight gain were observed as compared to control.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

effects observed, treatment-related

Description (incidence and severity):

Decreased numbers of fetuses/dam were observed in 3 mg/kg/day treated rat.

Reproductive function: sperm measures:

not specified

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

Pre- and postimplantation fetal mortality and decreased numbers of fetuses/dam were reported at the mid-dose

Mortality:
No mortality was observed in treated rat as compared to control.

Dose descriptor:

LOAEL

Effect level:

3 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Effect on weight gain, hematology, clinical chemistry, oxygen consumption, organ weight, gross pathology, histopathology and reproductive performance

Remarks on result:

other: Significant effect were observed

Dose descriptor:

NOAEL

Effect level:

0.3 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No adverse effect on survival, body weight gain, hematology, clinical, chemistry, gross pathology, histopathology and reproductive performance

Remarks on result:

other: No significant effect were observed at this dose.

Critical effects observed:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Mortality:
Pre and postimplantation fetal mortality were observed in 3 mg/kg/day treated rat.

Dose descriptor:

LOAEL

Generation:

F1

Effect level:

3 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Effect on survival of fetus

Remarks on result:

other: Effect were observed

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

0.3 mg/kg bw/day

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: No significant effect were observed

Remarks on result:

other: No significant effect were observed at this dose

Critical effects observed:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Reproductive effects observed:

not specified

Conclusions:

The NOAEL and LOAEL was considered to be 0.3 mg/kg/day and 3 mg/kg/day for F0 generation respectively and LOAEL was considered to be 3 mg/kg/day for F1 generation when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months.

Executive summary:

In a chronic reproductive toxicity study, white male and female rats were exposed to 2, 4, 6 trichloroaniline in the concentrations of 0, 0.3, 3 and 29 mg/kg/day by oral gavage. The results showed that 2, 4, 6 trichloroaniline is toxic at higher doses. Toxic changes were observed as decreased in weight gain,increased numbers of hypochromic RBCs and doubled levels of methemoglobin, Anisocytosis, poikilocytosis, reticulocytosis, hypochromia and presence of Heinz bodies in the RBCs and decreased in SDH and LDH activities in liver and the kidney of 29 mg/kg/day treated rat. Increased relative weights of brain and decreased liver weight. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain in 29 mg/kg/day treated rats. Decreased numbers of fetuses/dam and massive hematomas were observed in abdominal cavities of 3 mg/kg/day treated rat of F0 generation. In F1 generation, Pre and postimplantation fetal mortality were also observed in 3 mg/kg/day treated rat. Therefore, LOAEL was considered to be 3 mg/kg/day for F0 and F1 generation and NOAEL was considered to be 0.3 mg/kg/day for F0 generation when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months. This study is limited by inadequate data reporting, including the strain, size, and sex distribution of the control and treatment groups, the methods utilized,and the endpoints evaluated.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

0.3 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

Klemish 4 publication.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity:

In various experimental studies, 2,4,6-trichloroaniline (634-93-5)has been investigated for reproductive toxicity. The studies are mention below

In a chronic reproductive toxicity study, white male and female rats were exposed to 2, 4, 6 trichloroaniline in the concentrations of 0, 0.3, 3 and 29 mg/kg/day by oral gavage. The results showed that 2, 4, 6 trichloroaniline is toxic at higher doses. Toxic changes were observed as decreased in weight gain,increased numbers of hypochromic RBCs and doubled levels of methemoglobin, Anisocytosis, poikilocytosis, reticulocytosis, hypochromia and presence of Heinz bodies in the RBCs and decreased in SDH and LDH activities in liver and the kidney of 29 mg/kg/day treated rat. Increased relative weights of brain and decreased liver weight. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain in 29 mg/kg/day treated rats. Decreased numbers of fetuses/dam and massive hematomas were observed in abdominal cavities of 3 mg/kg/day treated rat of F0 generation. In F1 generation, Pre and postimplantation fetal mortality were also observed in 3 mg/kg/day treated rat. Therefore, LOAEL was considered to be 3 mg/kg/day for F0 and F1 generation and NOAEL was considered to be 0.3 mg/kg/day for F0 generation when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months. This study is limited by inadequate data reporting, including the strain, size, and sex distribution of the control and treatment groups, the methods utilized,and the endpoints evaluated.

In a multi generation study ,test chemical was administrated to male and female rats by drinking water at the concentration of 0, 2.5, 10 and 40mg/kg/day . Rats were continuously exposed to test chemical in the drinking water, beginning with birth of F0 generation and continuing through weaning of the F2 generation. The animals were observed for clinical sign, body weight, water intake fertility, growth, viability, locomotor activity, or blood chemical analyses .The treatment did not affect fertility, growth, viability, locomotor activity, or blood chemical analyses. Adrenal gland enlargement was observed in both the F0 generation and F1 animals at 95 d of age. Therefore NOAEL was considered to be 2.5mg/kg/day forP0 generation in male and female rats for test chemical multi generation study.

The aim of the no GLP-compliant study was to evaluate the potential genotoxic effect on germ cells in rat with a Dominant lethal test on rats. This test is a part of a combined study with 90 days repeated toxicity feeding test and teratogenicity test. After 90 days exposure period to test item mixed in food (0.1, 0.25 and 1% in food), males rats (20 per dose group) were mated with 340 young virgin females. Before mating procedure, male were treated with food for 19 weeks including 90 days study. Females were sacrified at day 17 of gestation. Observations were daily performed for conditions (live, dead, moribund) and weighed for males and females. After females sacrifice, corpora lutea were counted, numbers of implantation sites, resorptions sites and live and dead fetuses were counted. No difference between treated groups and control group was observed on living and dead fetuses ratios. Only bodyweight of treated animals (1%) was decresed and was lower than control.

Based on the data available for the target chemical 2,4,6-trichloroaniline (634-93-5)as well as applying weight of evidence does exhibit reproductive toxic nature and hence is likely to classify as per the criteria mentioned in CLP regulation in category 2. 

Effects on developmental toxicity

Description of key information

A devlopmental NOAEL was considered to be 0.3 mg/kg/day for F1 generation when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from secondary source .

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

A chronic reproductive toxicity study of 2, 4, 6-trichloroaniline was conducted in white male and female rats by oral gavage.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

not specified

Route of administration:

oral: gavage

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

Not specified.

Duration of treatment / exposure:

6 months

Frequency of treatment:

5days/week

Duration of test:

6 months

Remarks:

0.4, 4, or 40 mg/kg-day, 5 days/week, for 6 months
(adjusted to 0.3, 3, or 29 mg/kg-day)

No. of animals per sex per dose:

120 males and 60 females

Control animals:

yes, concurrent vehicle

Details on study design:

Not specified

Maternal examinations:

Clinical sign, histopathology were observed .

Ovaries and uterine content:

ovogenesis, (specific endpoints evaluated and methods utilized were not further specified)

Fetal examinations:

embryotoxicity, and teratogenicity were assessed(specific endpoints evaluated and methods utilized were not further specified)

Statistics:

Not specified

Indices:

Morphofunctional indices were observed.

Historical control data:

Not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

Pre- and postimplantation fetal mortality and decreased numbers of dam were reported at the mid-dose(3 mg/kg/day) compare to control.

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Description (incidence and severity):

The authors indicated that there were no significant variations in the “morphofunctional” indices (endpoints not specified) for the male and female reproductive organs.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Massive hematomas were observed in the abdominal cavities of mid-dose (3 mg/kg/day) adult rats. The researchers did not indicate whether the effects reported for mid-dose
rats also(3 mg/kg/day) occurred in high-dose(29mg/kg/day) rats.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Number of abortions:

not specified

Pre- and post-implantation loss:

not specified

Total litter losses by resorption:

not specified

Early or late resorptions:

not specified

Dead fetuses:

effects observed, treatment-related

Description (incidence and severity):

Pre- and postimplantation fetal mortality and decreased numbers of fetuses were reported at the mid-dose(3 mg/kg/day) compare to control.

Changes in pregnancy duration:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified

Changes in number of pregnant:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

0.3 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: No significant effect were observed at this dose .

Remarks on result:

other: No toxic effect were observed

Abnormalities:

not specified

Localisation:

not specified

Description (incidence and severity):

not specified

Dose descriptor:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

Abnormalities:

not specified

Localisation:

other: not specified

Description (incidence and severity):

not specified

Key result

Developmental effects observed:

yes

Lowest effective dose / conc.:

3 mg/kg bw/day

Treatment related:

yes

Relation to maternal toxicity:

not specified

Dose response relationship:

not specified

Relevant for humans:

no

Conclusions:

A devlopmental NOAEL was considered to be 0.3 mg/kg/day for F1 generation when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months.

Executive summary:

In a chronic reproductive toxicity study, white male and female rats were exposed to 2, 4, 6 trichloroanilinein the concentrations of 0, 0.3, 3 and 29 mg/kg/day by oral gavage. The results showed that 2, 4, 6 trichloroaniline is toxic at higher doses. Toxic changes were observed as decreased in weight gain,increased numbers of hypochromic RBCs and doubled levels of methemoglobin, Anisocytosis, poikilocytosis, reticulocytosis, hypochromia and presence of Heinz bodies in the RBCs and decreased in SDH and LDH activities in liver and the kidney of 29 mg/kg/day treated rat. Increased relative weights of brain and decreased liver weight. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain in 29 mg/kg/day treated rats. Decreased numbers of fetuses/dam and massive hematomas were observed in abdominal cavities of 3 mg/kg/day treated rat of P0 generation. In F1 generation, Pre and postimplantation fetal mortality were also observed in 3 mg/kg/day treated rat. No teratogenic effect were observed at this dose .Therefore, NOAEL was considered to be 0.3 mg/kg/day in F1 generation,when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months. This study is limited by inadequate data reporting, including the strain, size, and sex distribution of the control and treatment groups, the methods utilized,and the endpoints evaluated. Furter studies has to conducted to classify this substance.                                  

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

0.3 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Klemish 4 publication.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

In experimental study, 2,4,6-trichloroaniline (634-93-5)has been investigated for devlopmental toxicity. The study is mention below

In a chronic reproductive toxicity study, white male and female rats were exposed to 2, 4, 6 trichloroanilinein the concentrations of 0, 0.3, 3 and 29 mg/kg/day by oral gavage. The results showed that 2, 4, 6 trichloroaniline is toxic at higher doses. Toxic changes were observed as decreased in weight gain,increased numbers of hypochromic RBCs and doubled levels of methemoglobin, Anisocytosis, poikilocytosis, reticulocytosis, hypochromia and presence of Heinz bodies in the RBCs and decreased in SDH and LDH activities in liver and the kidney of 29 mg/kg/day treated rat. Increased relative weights of brain and decreased liver weight. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain in 29 mg/kg/day treated rats. Decreased numbers of fetuses/dam and massive hematomas were observed in abdominal cavities of 3 mg/kg/day treated rat of P0 generation. In F1 generation, Pre and postimplantation fetal mortality were also observed in 3 mg/kg/day treated rat. No teratogenic effect were observed at this dose .Therefore, NOAEL was considered to be 0.3 mg/kg/day in F1 generation,when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months. This study is limited by inadequate data reporting, including the strain, size, and sex distribution of the control and treatment groups, the methods utilized,and the endpoints evaluated. Furter studies has to conducted to classify this substance.                                  

Justification for classification or non-classification

Thus based on the above annotation for the target chemical 2,4,6-trichloroaniline (634-93-5) does exhibit reproductive toxic  and teratogenic nature and hence is likely to classify as per the criteria mentioned in CLP regulation in category 2.