2-amino-4-nitrophenol

Repeated dose toxicity: dermal

Administrative data

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data from handbook or collection of data

Data source

Reference

Reference Type:

publication

Title:

Teratology and percutaneous toxicity studies on hair dyes

Author:

C. Burnett

Year:

1976

Bibliographic source:

Journal of Toxicology and Environmental Health

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

Thirteen week study was conducted to evaluate the toxic effects of repeated administration of test chemical.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2-amino-4-nitrophenol

EC Number:

202-767-9

EC Name:

2-amino-4-nitrophenol

Cas Number:

99-57-0

Molecular formula:

C6H6N2O3

IUPAC Name:

2-amino-4-nitrophenol

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not specified

Administration / exposure

Type of coverage:

open

Vehicle:

other: 6% Hydrogen peroxide

Details on exposure:

TEST SITE
The sites of application on the dorsolateral aspects of the thoracic-lumbar area (one on each side of the midline) were alternated to minimize skin irritation.

REMOVAL OF TEST SUBSTANCE
The rabbits were restrained in holding stocks for 1 hr following each application and were then shampooed, rinsed, dried, and returned to their cages

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The test materials, prepared by mixing equal volumes of the dye solution and 6% hydrogen peroxide, were applied topically at a dose of 1000 mg/kg (
1 ml/kg) application.
USE OF RESTRAINERS FOR PREVENTING INGESTION: yes

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Twice weekly

Doses / concentrations

Remarks:

1000 mg/kg (1 ml/kg)

No. of animals per sex per dose:

Control – I: 6male and 6 females
Control - II : 6male and 6 females
Control - III: 6male and 6 females
2-Amino-4-nitrophenol : 6male and 6 females

Control animals:

yes, concurrent vehicle

Details on study design:

The hair at the site of application on the back and sides of each rabbit was clipped short throughou
t the study. The application sites on three animals of each sex in each group were abraded on the f
irst treatment day of each week. The rabbits were restrained in holding stocks for 1 hr following each
application and were then shampooed, rinsed, dried, and returned to their cages.

Examinations

Observations and examinations performed and frequency:

Observations and examinations performed and frequency
BODY WEIGHT: Yes,
The animals were weighed weekly during the study.
HAEMATOLOGY: Yes
Hematologic determinations was performed on all animals at 0, 3, 7, and 13 wk.
Following parameters was examined :
Blood count, Methemoglobin, Fasting blood sugar
CLINICAL CHEMISTRY: Yes
Clinical chemistry determinations was performed on all animals at 0, 3, 7, and 13 wk.
Following parameters was examined :
Blood urea nitrogen, Alkaline phosphatase, Serum glutamic oxaloacetic transaminase
URINALYSIS: Yes
Examination of urine was performed on all animals at 0, 3, 7, and 13 wk.
Urine was examined for color, pH, albumin, glucose, occult blood, and microscopic elements.
OTHER: Organ weights was determined.

Sacrifice and pathology:

All survivors were sacrificed after 13 wk and examined for gross abnormalities. Organ-body weight ra
tios were determined for liver,kidneys, adrenals, heart, thyroid, spleen, and brain.
Twenty-five tissues were examined microscopically.
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

Statistical analysis of the data on body weight gains, hematology, clinical chemistry, and absolute and relative organ weights was performed using the analysis of variance F test and Student's t test.When variances differed significantly, Student's t test was modified (t')and Cochran's approximation was utilized.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

No significant clinical sign was obsrved at dose level of 1000 mg/kg of treated group compare to control.

Dermal irritation:

not specified

Mortality:

mortality observed, non-treatment-related

Description (incidence):

Five control and five test animals died during the study due to complications resulting from cardiac puncture while collecting blood.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No significant clinical sign was obsrved at dose level of 1000 mg/kg of treated group compare to control.

Haematological findings:

no effects observed

Description (incidence and severity):

No effects (There were scattered statistically significant differences in the hematologic values between test and control groups at the various sampling intervals. However, these differences were
not considered to be of toxicologic significance because of either the direction or continuity of the differences or the fact that they fell within the range of historical control values.)

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

There were a few instances when there were statistically significant differences in relative organ weights between a test group and the combined controls but the differences were not significant when they were compared with each control group separately.

Gross pathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No microscopic lesions were seen that were judged to be due to the administration of the hair dye formulations.

Effect levels

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No significant effect were observed at this dose

Remarks on result:

other: No toxic effects were observed

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 1000 mg/kg/day for test chemical in New Zealand White male and female rabbit for 13 weeks by dermal route.

Executive summary:

Thirteen week study was conducted to evaluate the toxic effects of test substance. Test substance was applied dermally twice weekly for 13 week to groups of 12 adult New Zealand white male and female rabbits at a dose concentration of 1000 mg/kg. The animals were observed for dermal irritation,clinical sign, mortality, body weight, organ weight, gross and histopathology.There was no evidence of induced systemic effects. No effects on body weight. Microscopic examination of 25 tissues from each animal gave no indication of histomorphologic evidence of toxicity. No gross abnormalities were seen at necropsy. No dye discoloration of urine was seen at any time during the test or at necropsy.

 Therefore NOAEL of the study was considered to be 1000 mg/kg for test chemical in New Zealand White male and female rabbit for 13 weeks by dermal route.