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Repeated Dose toxicity: Oral

In a Subchronic repeated dose toxicity study, male and female white rats were exposed to the test chemical in the concentrations of 0, 80, 160 and 800 mg/kg/day. The test chemical was given by oral route to test animals for 45 days. The animals were observed for clinical signs, mortality changes in body weight, food consumption, organ weight, hemotological and clinical chemistry changes and the animals were subject to gross and histo-pathology. The results showed that 2, 4, 6 trichloroaniline is toxic at higher doses. Toxic changes were observed as depression, cyanosis, hair loss, and hematuria, lag in weight gain,reduction in hemoglobin and total number of red blood cells (RBCs), Polychromaphilic and hypochromic RBCs, signs of anisocytosis, poikilocytosis, and a tendency toward leucopenia and increased in alanine aminotransferase [ALT] and aspartate aminotransferase [AST], nitrogen and serum pyruvic acid levels and decreased ALT/AST ratio,rates of oxygen consumptioncatalase activity. SDH and LDH activities were inhibited in liver and the kidney of 800 mg/kg/day treated rat. Increase in relative weight of heart, liver, kidneys and spleen and decreased weight and volume of testicles were also observed. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain and histopathological changes such as tubules with desquamated spermatogenic epithelium in testicles in male rat were observed. Based on the observations made, the NOAEL (No observed adverse effect level) and low observed adverse effect level (LOAEL) was considered to be 80 mg/kg/day and 160 mg/kg/day respectively when rats were treated with the test chemical orally for 45 days.

Repeated dose toxicity: Inhalation

2,4,6-trichloroaniline  (634-93-5) has very low vapour pressure (0.0000195 Pa. = 1.4626200729e-7 P mmHg), So the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore the end point for repeated dose toxiciity by the inhalation route is considered for waiver.

Repeated dose toxicity: Dermal

Considering the use of 2,4,6 Trichloroaniline as an intermediate, repeated exposure of humans by the dermal route is unlikely. Also, The acute dermal toxicity value for 2,4,6 trichloroaniline (CAS no. 634-93-5) (as provided in section 7.2.3) is >2000 mg/kg body weight and hence this end point for repeated dose toxicity by dermal route is considered for waiver.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other:
Justification for type of information:
Data is available from secondary source
Qualifier:
according to guideline
Guideline:
other: Refer below principle
Principles of method if other than guideline:
Repeated dose toxicity study of the test chemical in rats was studied by oral administration
GLP compliance:
not specified
Species:
rat
Strain:
not specified
Details on species / strain selection:
No data
Sex:
male/female
Details on test animals or test system and environmental conditions:
No data
Route of administration:
oral: gavage
Vehicle:
other: Oil solution
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: 2,4,6-Trichloroaniline was mixed with 8% oil solution in the concentrtion of 0, 80, 160 and 800 mg/kg/day

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): oil solution were used
- Concentration in vehicle: 0, 80, 160 and 800 mg/kg/day
- Amount of vehicle (if gavage): 8%
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
45 days
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
0, 80, 160, or 800 mg/kg/day
Basis:
no data
No. of animals per sex per dose:
128 rats of both sexes
Control animals:
yes, concurrent vehicle
Details on study design:
No data
Positive control:
No data
Observations and examinations performed and 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: Before treatment and on days 10, 20, 30 and 45 of treatment.

BODY WEIGHT: Yes
- Time schedule for examinations: Before treatment and on days 10, 20, 30 and 45 of treatment.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY: No data available
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Before treatment and on days 10, 20, 30 and 45 of treatment.
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: All 128 treated rats were examined.
- Parameters checked in table [No.?] were examined: Concentration of formed elements and serum hemoglobin, Polychromaphilic and hypochromic RBCs and WBC were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Before treatment and on days 10, 20, 30 and 45 of treatment.
- Animals fasted: No data available
- How many animals: All 128 treated rats 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) and succinic dehydrogenase (SDH) activities in the liver and kidney were examined.

URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined. No data available

NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data available

OTHER:
Other parameters EKG at lead II, oxygen consumption and organ weight were examined.

Organ weighted:
Heart, liver, kidneys, and spleen, brain and testicles were examined.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Degenerative changes and volume of organ were observed.

HISTOPATHOLOGY: Yes

Organ examined: Heart, liver, kidneys, and spleen, brain and testicles were examined.
Other examinations:
No data available
Statistics:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
When treated with 800 mg/kg/day sign of toxicity such as depression, cyanosis, hair loss and hematuria were observed in treatecd rat as compare to control.
Mortality:
mortality observed, treatment-related
Description (incidence):
No mortality was observed in treated rat
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
A lag in body weight gain was observed in treated rat as compared to controls.
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:
effects observed, treatment-related
Description (incidence and severity):
The concentration of hemoglobin in the blood (Day 45) and the total number of red blood cells (RBCs) were statistically significantly (p < 0.05) reduced in the 800mg/Kg/day dosed group (approximately 25 and 27% less than controls). Polychromaphilic and hypochromic RBCs, signs of anisocytosis, poikilocytosis, and a tendency toward leucopenia were also noted
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
The activities of ALT and AST in the serum were increased approximately by about 45% and 20%, respectively, and the ALT/AST ratio was decreased in high-dose rats compared to controls. Levels of residual serum nitrogen and serum pyruvic acid were statistically significantly (p < 0.05) increased, and rates of oxygen consumption and serum catalase activity were statistically significantly (p < 0.02) decreased in the high-dose group of 800 mg//Kg/day
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
When treated with 800 mg/kg/day, increased in relative weight of heart, liver, kidneys and spleen and decreased weight and volume of testicles were observed in treated rat as compare to control. Decreased weight and volume of the testicles were noted in high-dose (800 mg/Kg/day) animals.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain were observed in 800 mg/kg/day treated rats.
Histopathological findings: non-neoplastic:
not specified
Description (incidence and severity):
Histological alterations were noted in the testicles (increased incidence of tubules with desquamated spermatogenic epithelium), but not the ovaries, of high-dose 800 mg/Kg/day rats. Similar, but less pronounced, evidence of toxicity was observed in 160 mg/kg/day dose rats.
Histopathological findings: neoplastic:
not specified
Dose descriptor:
NOAEL
Effect level:
80 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effect observed on hematology, clinical chemistry, organ weight, gross pathology and histopathology. There was no moartality observed.
Dose descriptor:
LOAEL
Effect level:
160 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Effect on clinical sign, hematology, clinical chemistry, organ weight, gross pathology and histopathology.
Critical effects observed:
not specified

Haematology:

When treated with 800 mg/kg/day, statistically significantly reduction in hemoglobin and total number of red blood cells (RBCs) was observed as compared to control.

 

Polychromaphilic and hypochromic RBCs, signs of

anisocytosis, poikilocytosis, and a tendency toward leucopenia were in treated rat

 

Parameter

Dose in mg/kg-day

Control

800

Hematology (means ± standard deviation)

Concentration of hemoglobin on Day 45 (g%)

15.88 ± 0.82

12.02 ± 2.08c

Number of RBCs (millions)

6.38 ± 0.25

4.63 ± 0.79d

 

c -Significantly different from control at p < 0.02

d- Significantly different from control at p < 0.001

 

Clinical chemistry:

When treated with 800 mg/kg/day, alanine

aminotransferase [ALT] and aspartate aminotransferase [AST], nitrogen and serum pyruvic acid levels in serum increased and ALT/AST ratio,rates of oxygen consumptioncatalase activity was decreased as compared to control.

SDH and LDH activities were inhibited in liver and the kidney of treated rat.

Parameter

Dose in mg/kg-day

Control

800

Clinical Chemistry (means ± standard deviation)

ALT (mmole)

2.57 ± 0.37

4.69 ± 0.5d

AST (mmole)

2.95 ± 0.27

3.74 ± 0.45d

Residual serum nitrogen(mg%)

34 ± 2.4

45.5 ± 6c

Serum pyruvic acid (mg%)

1.67 ± 0.1

2.36 ± 0.32d

Catalasee activity (mg%)

1.04 ± 0.11

0.18 ± 0.13d

 

c -Significantly different from control at p < 0.02

d- Significantly different from control at p < 0.001

Conclusions:
The NOAEL (No observed adverse effect level) and low observed adverse effect level (LOAEL) was considered to be 80 mg/kg/day and 160 mg/kg/day respectively when rats were treated with 2, 4, 6 trichloroaniline orally for 45 days.
Executive summary:

In a Subchronic repeated dose toxicity study, male and female white rats were exposed to 2, 4, 6 trichloroaniline in the concentrations of 0, 80, 160 and 800 mg/kg/day. The test chemical was given by oral route to test animals for 45 days. The animals were observed for clinical signs, mortality changes in body weight, food consumption, organ weight, hemotological and clinical chemistry changes and the animals were subject to gross and histo-pathology. The results showed that 2, 4, 6 trichloroaniline is toxic at higher doses. Toxic changes were observed as depression, cyanosis, hair loss, and hematuria, lag in weight gain,reduction in hemoglobin and total number of red blood cells (RBCs), Polychromaphilic and hypochromic RBCs, signs of anisocytosis, poikilocytosis, and a tendency toward leucopenia and increased in alanine aminotransferase [ALT] and aspartate aminotransferase [AST], nitrogen and serum pyruvic acid levels and decreased ALT/AST ratio,rates of oxygen consumptioncatalase activity. SDH and LDH activities were inhibited in liver and the kidney of 800 mg/kg/day treated rat. Increase in relative weight of heart, liver, kidneys and spleen and decreased weight and volume of testicles were also observed. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain and histopathological changes such as tubules with desquamated spermatogenic epithelium in testicles in male rat were observed. Based on the observations made, the NOAEL (No observed adverse effect level) and low observed adverse effect level (LOAEL) was considered to be 80 mg/kg/day and 160 mg/kg/day respectively when rats were treated with 2, 4, 6 trichloroaniline orally for 45 days.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
80 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
It is a recent report of 2010 and hence considered for end point selection

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: dermal
Data waiving:
other justification
Justification for data waiving:
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated Dose toxicity: Oral

Data available for the test chemicals was reviewed to determine the toxic nature of 2,4,6 trichloroaniline. The studies are as mentioned below:

In a Subchronic repeated dose toxicity study, male and female white rats were exposed to the test chemical in the concentrations of 0, 80, 160 and 800 mg/kg/day. The test chemical was given by oral route to test animals for 45 days. The animals were observed for clinical signs, mortality changes in body weight, food consumption, organ weight, hemotological and clinical chemistry changes and the animals were subject to gross and histo-pathology. The results showed that 2, 4, 6 trichloroaniline is toxic at higher doses. Toxic changes were observed as depression, cyanosis, hair loss, and hematuria, lag in weight gain,reduction in hemoglobin and total number of red blood cells (RBCs), Polychromaphilic and hypochromic RBCs, signs of anisocytosis, poikilocytosis, and a tendency toward leucopenia and increased in alanine aminotransferase [ALT] and aspartate aminotransferase [AST], nitrogen and serum pyruvic acid levels and decreased ALT/AST ratio,rates of oxygen consumptioncatalase activity. SDH and LDH activities were inhibited in liver and the kidney of 800 mg/kg/day treated rat. Increase in relative weight of heart, liver, kidneys and spleen and decreased weight and volume of testicles were also observed. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain and histopathological changes such as tubules with desquamated spermatogenic epithelium in testicles in male rat were observed. Based on the observations made, the NOAEL (No observed adverse effect level) and low observed adverse effect level (LOAEL) was considered to be 80 mg/kg/day and 160 mg/kg/day respectively when rats were treated with the test chemical orally for 45 days.

In another chronic repeated dose toxicity study, male and female rats were exposed to the test chemical in the concentrations of 0, 0.3, 3 and 29 mg/kg/day for 6 months. The test chemical was given by oral route to test animals for 6 months. The animals were observed for clinical signs, mortality changes in body weight, food consumption, organ weight, hemotological and clinical chemistry changes and the animals were subject to gross and histo-pathology.

The results showed that the test chemical is toxic. 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 and Decreased in Oxygen consumption at 15 minutes and higher numbers of associations for conditioned reflexes were observed in 3 mg/kg/day compared to controls. Based on the observations made, the No observed adverse effect leve (NOAEL) and low observed adverse effect level (LOAEL) is considered to be 0.3 mg/kg/day and 3 mg/kg/day when rats were treated with the test chemical orally for 6 months.

In another study, chronic repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical. The study was performed using male and female rats. The test chemical was dissolved in suitable solvent and dosed to test animals by gavage route for 4 months at dose level of0, 5, 20 and 100 mg/kg/day. During the study period, the animals were observed for growth, changes in food consumption, hematology and clinical chemistry parameters. Haematological examinations (RBC, total and differentialleucocyte, platelet, haematocrit and haemoglobin concentration), blood sugar, as well as growth and food consumption data indicated no significant effects attributable to 2,6-dichloro-4-nitroaniline at any of the dose levels or treatments. Based on the observations made, the No observed adverse effect level (NOAEL) for the test chemical is considered to be 100 mg/Kg/day.

Repeated dose toxicity: Inhalation

2,4,6-trichloroaniline  (634-93-5) has very low vapour pressure (0.0000195 Pa. = 1.4626200729e-7 P mmHg), So the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore the end point for repeated dose toxiciity by the inhalation route is considered for waiver.

Repeated dose toxicity: Dermal

Considering the use of 2,4,6 Trichloroaniline as an intermediate, repeated exposure of humans by the dermal route is unlikely. Also, The acute dermal toxicity value for 2,4,6 trichloroaniline (CAS no. 634-93-5) (as provided in section 7.2.3) is >2000 mg/kg body weight and hence this end point for repeated dose toxicity by dermal route is considered for waiver.

Based on the observations made, the test chemical 2,4,6 -trichloroaniline is not likely to be a toxicant as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the observations made, the test chemical 2,4,6 -trichloroaniline (CAS no 634 -93 -5) is not likely to be a toxicant as per the criteria mentioned in CLP regulation.