Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 14 November to 5 December 1979
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a reliable method and to GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report Date:
1981

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EPA OPP 82-2 (Repeated Dose Dermal Toxicity -21/28 Days)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Batch No.: 52829055
Purity: 99.5%

Test animals

Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Langshaw Farms, Augusta, Michigan
- Age at study initiation:
- Weight at study initiation: 2127 to 2891 grams
- Fasting period before study:
- Housing: Individually housed in hanging wire-mesh cages.
- Diet (e.g. ad libitum): Purina Certified Rabbit Chow, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 70-76 °F
- Humidity (%): 44-57%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark

IN-LIFE DATES: From: 14 November 1979 To: 5 December 1979

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
physiological saline
Remarks:
0.9%
Details on exposure:
TEST SITE
- Area of exposure: 100 cm2
- % coverage: 23%, 35% and 100%
- Type of wrap if used: Each site was covered with a 10 x 10-centimeter gauze pad with adhesive tape borders. The entire area was then wrapped with gauze bandaging and Elastoplast tape.
- Time intervals for shavings or clipplings: twice each week prior to test article administration during the three week study period

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The test areas were washed with tepid IRDC tap water and dried with disposable paper towels.
- Time after start of exposure: 6 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100, 500 and 2500 mg/kg bw
- For solids, paste formed: no

VEHICLE
not applicable

USE OF RESTRAINERS FOR PREVENTING INGESTION: no
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
6 hours, test substance moistened with physiological saline
Frequency of treatment:
Once a day, 5 days per week, for 3 consecutive weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
100, 500, 2500 mg/kg
Basis:
nominal per unit body weight
No. of animals per sex per dose:
4 males and 4 females per dose
Control animals:
yes
Details on study design:
None stated
Positive control:
None stated

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily during the 21-day study period
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once daily

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: once daily

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were obtained during the pretest period, prior to study initiation and at weekly intervals during the 21-day study period

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data

WATER CONSUMPTION: No data

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: once during the pretest period and at 21 days of study
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: all animals
- Parameters: hematocrit, hemoglobin, erythrocyte count, total leucocyte count, differential leucocyte count, platelets, MCH, MCV and MCHC

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Once during the pretest period and at 21 days of study.
- Animals fasted: Yes / No / No data
- How many animals: all animals
- Parameters:glucose, blood urea nitrogen, lkaline phosphatase, serum glutamic pyruvic transaminase, and serum glutamic oxalacetic transaminase

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data

OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
masses and lesions, untreated skin, treated skin, kidney, lungs, pituitary, sciatic nerve, muscle, spleen, urinary bladder, thymus, lumbar spinal cord, pancreas, stomach, duodenum, testes, thyroid, femur, colon, mesenteric lymph nodes, mediastinal lymph nodes, ovaries, parathyroid, liver, gall bladder, adrenals, heart, brain
HISTOPATHOLOGY: Yes
spleen, pancreas, stomach, duodenum, colon, lymph nodes, urinary bladder, heart, lung, brain (3 sections), treated skin (3 sections), untreated skin (3 sections), liver, kidney, gall bladder, adrenals, spinal cord, gonads, nerve (with muscle), bone marrow, thyroid (parathyroid), pituitary, thymus
Other examinations:
None stated
Statistics:
All statistical analyses compared the treatment groups with the control group, by sex.
Body weights (day 21), hematological and biochemical parameters (pretest and day 21) and absolute and relative organ weights (terminal sacrifice) were compared by analysis of variance (one-way classification), Bartlett’s test for homogeneity of variances and the appropriate t-test (for equal or unequal variances) as described by Steel and Torrie using Dunnett’s multiple comparison tables to judge significance of differences.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Dermal irritation:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
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:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not specified
Details on results:
CLINICAL SIGNS AND MORTALITY
In the control group, a few rabbits exhibited signs of anorexia and purulent nasal discharge. One rabbit also exhibited diarrhea.
In the test groups, a few rabbits exhibited signs of anorexia, purulent nasal discharge, clear nasal discharge, soft stool, diarrhea and ocular discharge.
None of the rabbits died during the 21-day study period.

DERMAL IRRITATION
Very slight dermal irritation was noted for one rabbit at the 2500 mg/kg dosage level.

BODY WEIGHT AND WEIGHT GAIN
There were no statistical differences found between the control and test groups.

HAEMATOLOGY
There were no treatment-related differences noted in the hematology data in this study.

CLINICAL CHEMISTRY
Although there was one statistically significant difference noted, this was not considered physiologically meaningful or treatment-related. The mid-dose female SGOT mean was significantly lower than the control mean, however, there were only four females in each group and the difference was only 5 IU/1. Therefore the statistically significant difference observed does not have any biological significance.

ORGAN WEIGHTS
The comparison of the control and treated group means for body and organ weights did not show any important differences.

GROSS PATHOLOGY
The skin from the application site was described as erythematous in one, two and seven animals of groups I, III and IV, respectively. The lungs presented foci or areas of congestion that were considered unrelated to the application of the test compound.

HISTOPATHOLOGY: NON-NEOPLASTIC
The skin from the application site presented mainly infiltration of inflammatory cells in the dermis with slight severity in both control and treated groups. Slight hyperkeratosis was observed in three males and four females in group IV. These lesions were very slight and represent a minimal reaction to the test compound.
The foci of myocarditis found in some treated animals were considered unrelated to the application of the test compound since they are observed from time to time in control animals.

Effect levels

Dose descriptor:
NOAEL
Effect level:
> 2 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: One female in the 2500 mg/kg test group exhibited dermal irritation during the 21-day study period.

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The NOAEL was considered to be > 2500 mg/kg. Very slight dermal irritation was noted for one rabbit at this dosage level. No compound related changes were seen in general behavior and appearance, body weight, clinical laboratory tests, organ weights or survival.
Executive summary:

In a dermal study in New Zealand White rabbits, the test substance was applied 5 days a week for 3 weeks at dosage levels of 100, 500 and 2500 mg/kg. Four male and four female rabbits were used in each treated group and in a control group. 0.9% physiological saline was used as the control substane. The rabbits were observed daily for signs of dermal irritation and changes in general behavior and appearance. Individual body weights were recorded weekly Hematologic and biochemical studies were conducted once in the pretest period and at 21 days of study.

The NOAEL was considered to be > 2500 mg/kg. Very slight dermal irritation was noted for one rabbit at this dosage level. No compound related changes were seen in general behavior and appearance, body weight, clinical laboratory tests, organ weights or survival.