Registration Dossier

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:
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

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals 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.