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:
other: Data is from peer reviewed journal

Data source

Reference
Reference Type:
publication
Title:
TERATOLOGY AND PERCUTANEOUS TOXICITY STUDIES ON HAIR DYES
Author:
C. Burnett, E. I. Goldenthal, S. B. Harris, F. X. Wazeter, J. Strausburg, R. Kapp, R. Voelker
Year:
1976
Bibliographic source:
Journal of Toxicology and Environmental Health, 1:6, 1027-1040, 1976

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other:
Principles of method if other than guideline:
Subchronic dermal toxicity study of 4-Chlororesorcinol (as part of mixture 7406) in rabbits
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
4-chlororesorcinol
EC Number:
202-462-0
EC Name:
4-chlororesorcinol
Cas Number:
95-88-5
Molecular formula:
C6H5ClO2
IUPAC Name:
4-chlorobenzene-1,3-diol
Details on test material:
- Name of test material (as cited in study report): 4-Chlororesorcinol
- Molecular formula (if other than submission substance): C6H5O2Cl
- Molecular weight (if other than submission substance): 144.56 g/mol
- Substance type: Organic
- Physical state: Liquid
- Impurities (identity and concentrations): No data available

Test animals

Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Housing: Animals was housed in cages

Administration / exposure

Type of coverage:
open
Vehicle:
other: Hydrogen peroxide
Details on exposure:
TEST SITE
- Area of exposure: Dorsolateral aspects of thoracic-lumbar area (one on each side of the midline) were exposed.
- % coverage: No data available
- Type of wrap if used: No data available
- Time intervals for shavings or clipplings: The hair at the site of application on the back and sides of each rabbit was clipped short throughout the study.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The rabbits were shampooed, rinsed and dried
- Time after start of exposure: 1 hr

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 1 ml/kg
- Concentration (if solution): 1 ml/kg of 1:1 oxidation mixture were used.
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- Justification for use and choice of vehicle (if other than water): Hydrogen peroxide was used
- Amount(s) applied (volume or weight with unit): No data available
- Concentration (if solution): 6% hydrogen peroxide.
- Lot/batch no. (if required): No data available
- Purity: No data available

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes
The rabbits were restrained in holding stocks for 1 hr following each application
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Twice weekly (The application sites on three animals of each sex in each group were abraded on the first treatment day of each week).
Doses / concentrations
Remarks:
Doses / Concentrations:
1 ml/kg (equivalent to 340 mg/kg)
Basis:
no data
No. of animals per sex per dose:
Control 1 : 6 male, 6 female
Control 2 : 6 male, 6 female
Control 3 (combined control) : 6 male, 6 female
1 ml/kg : 6 male, 6 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: 1 ml/kg dose were applied as it was the maximum dose that could be applied on the side of the rabbits without runoff.
- Rationale for animal assignment (if not random): No data available
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked: Mortality was observed.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data available.

DERMAL IRRITATION (if dermal study): No data available.
- Time schedule for examinations: No data available.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION: 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

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: 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: Blood was collected on day 0, 3, 7and 13 of study.
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: All 48 animals were examined.
- Parameters checked in table [No.?] were examined: Blood count, methemoglobin, fasting blood sugar, HCT, Hb, RBC, WBC and Met-Hb were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood was collected on day 0, 3, 7and 13 of study.
- Animals fasted: No data available
- How many animals: All 48 animals were examined.
- Parameters checked in table [No.?] were examined: Glucose, BUN, Alkaline Phosphatase and SGOT were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: Urine was collected on day 0, 3, 7and 13 of study.
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined: Color, pH, albumin, glucose, occult blood, and microscopic elements were examined.

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: Organ weight was examined.

Organ weighed: Liver, kidneys, adrenals, heart, thyroid, spleen, brain, stomach, duodenum, colon, gall bladder, adrenals, eyes, pancreas, kidneys, urinary bladder, ovaries, testes, bone, bone marrow and lung were examined.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Gross abnormalities were examined by sacrificing all the treated animals at the termination of study.

HISTOPATHOLOGY: Yes
Organ was stained with hematoxylin and eosin and examined microscopically.
Organs examined: Skin from treated and untreated areas, lymph nodes (mesanteric, axillary, and cervical), spleen, stomach, duodenum, colon, liver, gall bladder, adrenals, nerve with adjacent muscle, eyes, pancreas, kidneys, urinary bladder, ovaries, testes, bone, bone marrow, heart, lung, thyroid, brain and skeletal muscle under the site of application and elsewhere (thigh) were examined
Other examinations:
No data available
Statistics:
Statistical analysis was performed using the analysis of variance F test and Student's t test for body weight gains, hematology, clinical chemistries, and absolute and relative organ weights. When variances differed significantly, Student's t test was modified (t’) and Cochran's approximation was used.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
No mortality were observed in treated rabbits as compared to control. Five control and five treated animals died during the study due to complications resulting from cardiac puncture while collecting blood for blood investigations. Clinical signs: No sign of toxicity was observed in treated rabbits as compared to control.
Dermal irritation:
not examined
Mortality:
no mortality observed
Description (incidence):
No mortality were observed in treated rabbits as compared to control. Five control and five treated animals died during the study due to complications resulting from cardiac puncture while collecting blood for blood investigations. Clinical signs: No sign of toxicity was observed in treated rabbits as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No body weight and weight gain was observed in treated rabbits as compared to control.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
In treated female rabbits, there was significant increase in RBC and significant decreased in Met-Hb and in male, significant decreased in Met-Hb was observed. However, these differences were not considered to be of toxicological significance.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Scattered statistically significant differences were observed in treated rabbits but were not considered to be of significance because of either the direction or continuity of the differences or because they fell in the range of historical control values.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No effect was observed in urine of treated rabbits as compared to control.
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
In treated rabbits, statistically significant differences in relative organ weight were observed as compared to combined control but the difference was not significant when compared with control.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No gross abnormalities were observed in treated animals
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No microscopic lesions were observed in treated rabbits as compared to control.
Histopathological findings: neoplastic:
no effects observed
Details on results:
No gross abnormalities were seen at necropsy and no microscopic examination that were judged to be due to the administration of the test compound. The incidence and severity of disease processes common to laboratory rabbits was not affected by the experimental treatments.

Effect levels

Dose descriptor:
NOAEL
Effect level:
340 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on survival, clinical sign, body weight, hematology, clinical chemistry, urine analysis, organ weight, gross pathology and histopathology

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 340 mg/kg bw/day for New Zealand white male and female rabbits when treated with 4-Chlororesorcinol (as part of mixture 7406).
Executive summary:

In subchronic dermal repeated dose toxicity study, male and female New Zealand white rabbits were exposed to 4–Chlororesorcinol in the concentration of 0 and 1 ml/kg (equivalent to 340 mg/kg bw/day).

The results showed that 4 -Chlororesorcinol was not toxic. No effect were observed on survival, no evidences of percutaneous toxicity, no changes in body weight, hematology, clinical chemistry and absolut and relative organ weight of treated animals. In addition, no gross pathological and histopathological changes were observed in treated rabbits.

Therefore, NOAEL was considered to be 340 mg/kg bw/day when New Zealand white male and female rabbits were exposed to 4-Chlororesorcinol (as part of mixture 7406) dermally for 13 weeks.

This value indicates that the substance is unlikely to exhibit repeated dose toxicity by the dermal route at the above mentioned dose level.