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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
repeated dose toxicity: oral, other
Remarks:
Combined repeated dose & carcinogenicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Repeated dose oral toxicity study of the test chemical
Author:
Imaida et al
Year:
1983
Bibliographic source:
Toxicology Letters

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Refer below principle
Principles of method if other than guideline:
Combined repeated dose & carcinogenicity study was performed to determine the toxic nature of the test chemical
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: p-phenylenediamine- Molecular formula: C6H8N2- Molecular weight: 108.1432 g/mol- Substance type: Organic- Physical state: No data- Impurities (identity and concentrations): No data

Test animals

Species:
rat
Strain:
Fischer 344
Details on species / strain selection:
No data
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Charles River Japan, Inc., Kanagawa, Japan- Age at study initiation: 6 weeks- Weight at study initiation: No data- Fasting period before study: No data- Housing: Rats were housed in plastic cages- Diet (e.g. ad libitum): Oriental M powdered diet (Oriental Yeast Co., Tokyo, Japan)- Water (e.g. ad libitum): Tap water ad libitum- Acclimation period: No dataENVIRONMENTAL CONDITIONS- Temperature (°C): 24˚C- Humidity (%):No data- Air changes (per hr): No data- Photoperiod (hrs dark / hrs light): No dataIN-LIFE DATES: From: To: No data

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
No data
Vehicle:
other: Feed
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with feed at dose levels of 0, 25 or 50 mg/Kg/dayDIET PREPARATION- Rate of preparation of diet (frequency): No data- Mixing appropriate amounts with (Type of food): No data- Storage temperature of food: No dataVEHICLE- Justification for use and choice of vehicle (if other than water): No data- Concentration in vehicle: 0, 25 or 50 mg/Kg/day- Amount of vehicle (if gavage): No data- Lot/batch no. (if required): No data- Purity: No data
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
80 weeks
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
0, 25 or 50 mg/Kg/day
No. of animals per sex per dose:
0 mg/Kg/day: 24-25 rats25 mg/Kg/day: 63-66 rats50 mg/Kg/day: 63-66 rats
Control animals:
yes, concurrent vehicle
Details on study design:
Details on study design- Dose selection rationale: The concentrations of p-phenylenediamine used were decided from the results of the subacute toxicity test. - Rationale for animal assignment (if not random): No data- Rationale for selecting satellite groups: No data- Post-exposure recovery period in satellite groups: No data- Section schedule rationale (if not random): No data
Positive control:
No data

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No data- Time schedule: No data- Cage side observations checked in table [No.?] were included. No dataDETAILED CLINICAL OBSERVATIONS: No data- Time schedule: No dataBODY WEIGHT: Yes- Time schedule for examinations: weeklyFOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, weekly- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes - Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No dataFOOD EFFICIENCY:- 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 dataWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data- Time schedule for examinations: No dataOPHTHALMOSCOPIC EXAMINATION: No data- Time schedule for examinations: No data- Dose groups that were examined: No dataHAEMATOLOGY: Yes- Time schedule for collection of blood: All animals surviving in week 80 were subjected to hematological analysis.- Anaesthetic used for blood collection: No data - Animals fasted: No data - How many animals: All animals surviving in week 80- Parameters checked in table [No.?] were examined. No dataCLINICAL CHEMISTRY: No data - Time schedule for collection of blood: No data- Animals fasted: No data- How many animals: No data- Parameters checked in table [No.?] were examined. No dataURINALYSIS: No data - Time schedule for collection of urine: No data- Metabolism cages used for collection of urine: No data- Animals fasted: No data - Parameters checked in table [No.?] were examined. No dataNEUROBEHAVIOURAL EXAMINATION: No data- Time schedule for examinations: No data- Dose groups that were examined: No data- Battery of functions tested: sensory activity / grip strength / motor activity / other: No dataOTHER: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, Animals were killed after 80 weeks or when they became moribund and were necropsied after death. All organs were examinedMacroscopically.HISTOPATHOLOGY: Yes, All organs were examined macroscopically, and then fixed in 10% buffered formalin and sections were prepared for histological examination.
Other examinations:
No data
Statistics:
No data

Results and discussion

Results of examinations

Clinical signs:
not specified
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was no relation between the average body weight and the concentration of p-phenylenediamine in male rats, but the body weight of female rats given 0.1% p-phenylenediamine was less than that of the controls.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
The food consumption of male and female rats in the experimental groups was not significantly different from those of the controls.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
The number of red and white blood cells and the hematocrit and hemoglobin values were not significantly different in different groups.
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:
effects observed, treatment-related
Description (incidence and severity):
In females in group 1, given 0.1% p-phenylenediamine, the mean weights of the body and spleen were less than those of the control, and in females in group 2, given 0.05%, the mean weight of the spleen was less than that of the control.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
The incidences of non-neoplastic lesions, including hemorrhage of the pituitary gland, fatty degeneration of the liver, fibrosis of the pancreas and pneumonia, were also not significantly different in different groups. No marked changes of the thyroid gland were observed in any rats.
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
The first tumor (a pheochromocytoma) was observed in a male rat that died of pneumonia in week 60. In both sexes the highest incidence of neoplastic lesions was that of pheochromocytomas of the adrenal gland. These lesions were observed in 10 (27.8%) of 36 male rats given 0.1% p-phenylenediamine, 8 (22.9%) of 35 male rats given 0.05% p-phenylenediamine and 6 (31.6%) of 19 males in the control group, but there was no significant difference in their incidences in different groups. In females, pheochromocytomas were also found in all experimental groups, although at lower incidences than in males. Other neoplastic lesions were as follows: hyperplasia of the forestomach in males, a fibroadenoma of the mammary gland in a female, a fibroma of the skin in a male, lymphomas in females and ductal hyperplasia of the pancreas in a female. The incidences of these neoplastic lesions were not significantly different in different groups.
Other effects:
not specified

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
25 mg/kg bw (total dose)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: No significat effects were noted at the mentioned dose level
Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw (total dose)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: No significant alterations were noted at the mentioned dose level

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table: Hematological Data On Rats Treated For Up To 80 Weeks

Treatment (mg/Kg/day)

Sex

Number of rats

RBC (104/mm3)

WBC (/mm3)

Ht (%)

Hb (g/dl)

50

M

16

823.6±167.1b

42600±23900

43.6± 7.2

18.3± 2.1

25

M

11

1099.2± 82.0

59553± 22792

56.2± 6.4

21.9± 2.2

0

M

1

1016.0

12550

40.1

14.3

50

F

11

802.6±103.9

44570± 19271

44.5± 7.8

20.5±2.4

25

F

32

883.0± 158.0

46265 ±18398

47.0±9.3

19.0± 2.5

0

F

6

1077.9± 155.9

16550±1602

44.6±10.6

15.7± 3.5

 

Table: Weights of body, liver, kidney and spleen of rats treated for up to 80 weeks

Treatment (mg/Kg/day)

Sex

Number of rats

Terminal body weight (g)

Liver

Kidneys

Spleen

Left

Right

g

%bw

g

%bw

g

%bw

g

%bw

50

M

16

243.8±46.6b

5.74±0.91

(2.3)

1.07±0.10

 

(0.4)

1.08±0.11

(0.4)

0.50±0.11

(0.2)

25

M

11

207.8±29.5

5.29± 0.80

(2.5)

1.00±0.09

(0.5)

0.99±0.09

(0.5)

0.39±0.08

(0.2)

0

M

1

241.7

5.02

(2.1)

1.02

(0.4)

0.96

(0.4)

0.41

(0.2)

50

F

11

138.5±20.6

4.71±0.89

(3.4)

0.78±0.05

(0.6)

0.79±0.09

(0.6)

0.44±0.12e

(0.3)

25

F

32

171.5±31.8

3.97±0.65

(2.3)

0.69±0.05

(0.4)

0.68±0.04

(0.4)

0.46±0.14

 

(0.3)

0

F

6

174.4±2.5

3.57±0.24

(2.0)

0.56±0.16

(0.3)

0.71±0.01

(0.4)

0.63±0.02

(0.4)

 

 Table: Incidence Of Preneoplastic And Neoplastic Lesions In treated F344 Rats

Organ

Lesion

Number of rats affected/Number examined (%)

Female

Male

Dose (mg/Kg/day)

0

25

50

0

25

50

Adrenal gland

Pheochromocytoma

1/21

1/37

2/42

6/19

8/35

10/36

Forestomach

Hyperplasia

0/21

0/37

0/42

0/19

6/35

1/36

Mammary gland

Fibroadenoma

0/21

0/37

1/42

0/19

0/35

0/36

Skin

Fibroma

0/21

0/37

0/42

1/19

0/35

0/36

Hematopoietic system

Lymphoma

1/21

0/37

1/42

0/19

0/35

0/36

Pancreas Ductal

hyperplasia

0/21

0/37

1/42

0/19

0/35

0/36

 

Applicant's summary and conclusion

Conclusions:
The No Observed Adverse Effect Level (NOAEL) for the test chemical is considered to be 25 mg/kg/day for female rats and 50 mg/Kg/day for male rats when F344 rats were exposed to the test chemical for 80 weeks.
Executive summary:

Combined repeated dose & carcinogenicity study was performed to determine the toxic nature of the test chemical. The study was performed using 6 week old male F344 rats in a 80 weeks study. The test chemical was mixed with feed and used at dose level of 0, 25 or 50 mg/Kg/day.The animals were observed for changes in body weight and food consumption, hematology, organ weight changed and were subjected to gross and histopathology. There was no relation between the average body weight and the concentration of p-phenylenediamine in male rats, but the body weight of female rats given 0.1% p-phenylenediamine was less than that of the controls. The food consumption of male and female rats in the experimental groups was not significantly different from those of the controls. The number of red and white blood cells and the hematocrit and hemoglobin values were not significantly different in different groups. In females in group 1, given 0.1% p-phenylenediamine, the mean weights of the body and spleen were less than those of the control, and in females in group 2, given 0.05%, the mean weight of the spleen was less than that of the control. The first tumor (a pheochromocytoma) was observed in a male rat that died of pneumonia in week 60. In both sexes the highest incidence of neoplastic lesions was that of pheochromocytomas of the adrenal gland. These lesions were observed in 10 (27.8%) of 36 male rats given 0.1% p-phenylenediamine, 8 (22.9%) of 35 male rats given 0.05% p-phenylenediamine and 6 (31.6%) of 19 males in the control group, but there was no significant difference in their incidences in different groups. In females, pheochromocytomas were also found in all experimental groups, although at lower incidences than in males. Other neoplastic lesions were as follows: hyperplasia of the forestomach in males, a fibroadenoma of the mammary gland in a female, a fibroma of the skin in a male, lymphomas in females and ductal hyperplasia of the pancreas in a female. The incidences of these neoplastic lesions were not significantly different in different groups. The incidences of non-neoplastic lesions, including hemorrhage of the pituitary gland, fatty degeneration of the liver, fibrosis of the pancreas and pneumonia, were also not significantly different in different groups. No marked changes of the thyroid gland were observed in any rats. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for the test chemical is considered to be 25 mg/kg/day when F344 rats were exposed to the test chemical for 80 weeks.