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Toxicological information

Repeated dose toxicity: dermal

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Administrative data

Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1 to 30 April 1980
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
It is a study that has been published in a peer reviewed journal. The restriction is also due to the use of the read across approach: the test was performed not with 3-PPB but with Benzyl benzoate, a substance which has been demonstrated to be very similar in structure, physical/chemical properties and toxicological profile .

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1980
Report Date:
1980

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
Deviations:
no
Principles of method if other than guideline:
In principle the methods used are the same as those described in standard regulatory guidelines for subacute determination of dermal toxicity
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: clear liquid
Details on test material:
- Name of test material (as cited in study report): Benzyl benzoate

- Physical state: clear liquid
- Analytical purity: no data
-
- Storage condition of test material: amber glass bottle at room temperature
- Other: Specific gravity - 1.12

Test animals

Species:
rat
Strain:
other: CD SD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, USA
- Age at study initiation: no data
- Weight at study initiation: group mean bodyweights at study initiation was , males : 297-344g and females: 198-225g
- Fasting period before study: not applicable
- Housing: individually housed in stainless steel mesh cages
- Diet (e.g. ad libitum):Purina Rat Chow, supplied by Stover Feed Company, ad libitum
- Water (e.g. ad libitum): ad libitum from automatic watering system
- Acclimation period: 4 weeks

IN-LIFE DATES: From: 1 April 1980 To: 30 Apil 1980

Administration / exposure

Type of coverage:
open
Vehicle:
unchanged (no vehicle)
Details on exposure:
TEST SITE
- Area of exposure: not specified - seven shaved areas identified on the dorsum. Treatment of the prepared sites was rotated over each 7 day period
- % coverage: no information
- Type of wrap if used: none used
- Time intervals for shavings or clipplings: dorsum shaved before dosing commenced, no details for interim subsequent shaving occasions

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no details


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): six dose levels - 0.188; 0.301; 0.488; 0.781; 1.25 and 2.0 g/kg bw


USE OF RESTRAINERS FOR PREVENTING INGESTION
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
No information
Duration of treatment / exposure:
4 weeks
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0.188; 0.301; 0.488; 0.781; 1.25 and 2.0 g/kg bw
Basis:
nominal per unit body weight
No. of animals per sex per dose:
three males and three females per group
Control animals:
no
Details on study design:
Dose selection rationale: doses selected to provide range-finding information for a 90-day study
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: not applicable
Positive control:
No

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No data
DETAILED CLINICAL OBSERVATIONS: No data

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

BODY WEIGHT: Yes
- Time schedule for examinations: weekly recording

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

FOOD 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

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No


HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to termination, obtained by cardiac puncture
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes
- How many animals: all - 3 males and 3 females per group
- Parameters checked : haematocrit; haemoglobin; erythrocytes; total and differential leucocytes

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:immediately prior to termination, obtained by cardiac puncture
- Animals fasted: Yes
- How many animals:all - 3 males and 3 females per group
- Parameters checked : alkline phosphatase AP; lactic dehydrogenase; serum glutamic-oxalacetic transaminase GOT; serum glutamic-pyruvic transaminase GPT; glucose, cholesterol, calcium, phosphorus; uric acid; blood urea nitrogen, total protein, total bilirubin and albumin

URINALYSIS: Yes
- Time schedule for collection of urine: samples collected directly from under housing cages on day prior to or day of terminal sacrifice
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- Parameters checked : appearance, pH, ketones, occult blood, bilirubin, protein, glucose, microscopic examination of sediment
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
On day 31, all surviving rats were sacrificed following an overdose of ether . All animals including any decedents were examined for gross pathology. The following tissues were processed for histoptholoical examination:

treated and untreated skin
adrenals
heart
lung and bronchi
mesenteric lymph nodes
thyroid
pituitary
brain (3 sections)
sternal bone marrow
spinal cord
testes with epididymus
ovaries
spleen
urinary bladder
nerve with muscle
kidney
gross lesions
Other examinations:
no data
Statistics:
No data

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Dermal irritation:
no effects observed
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
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:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY:
No clinical signs of adverse reaction to dermal application of doses from 0.188 to 1.25 g/kg. In the high dose group three males and 1/3 females died between days 6 and 9. Prior to death the majority of these rats showed anogenital discharge, tremors and ataxia. One of the surviving females showed anogenital discharge and hyperactivity on days 7 to 9 but survived the episode, the second surviving female had tremors on days 6 to 8.

BODY WEIGHT AND WEIGHT GAIN:
For rats in the groups dosed at 0.188, 0.301, 0.488 or 0.781 g/kg, there were no effects on bodyweight or weight gain. Bodyweight gains in the group dosed at 1.25 g/kg were slightly lower than the other groups. In the high dose group, 2.0 g/kg, 4/6 rats died and bodyweight gains were not assessed for the remaining rats.

DERMAL REACTIONS:
The treated skin sites (seven areas on the dorsum dosed on a 7 day rotation) were not affected by repeated topical application of benzyl benzoate despite the recording of a few instances of slight redness or oedema.

HAEMATOLOGY:
No notable differences between groups for any of the blood parameters. White blood cell counts and differential counts in groups 5 and 6 were not significantly different (despite only two rats surviving at the highest dose level). Mean white cell counts for the treated groups 1-6 were 10.4; 11.9; 10.1; 11.2; 8.4 and 8.6 respectively.

CLINICAL CHEMISTRY:
No effects on any of the parameters except for a trend for higher uric acid levels in groups treated at the lower dose levels

URINALYSIS:
No effects of treatment were detected in any of the urinary parameters examined at any dose level.

GROSS PATHOLOGY:
All animals, including the decedents, showed no macroscopic abnormalities during necropsy.

HISTOPATHOLOGY-NON-NEOPLASTIC:
The histopathologically recognised effects of dermal application of benzyl benzoate to rats are slight squamous epithelial hyperplasia, degeneration of hair follicles and sebaceous glands, subcutaneous fibrosis and thyroid gland hyperplasia

Effect levels

Dose descriptor:
NOAEL
Effect level:
781 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Reduced weight gain at 1250 mg/kg bw/d; mortality at 2000 mg/kg bw/d

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
In a 30 day study, female and male rats received dermal application of 188, 301, 488, 781 and 1250 and 2000 mg/kg bw benzyl benzoate. 2000 mg/kg bw/d resulted in signs of toxicity and mortality; however effects at lower dose levels were limited to reduced weight gain at 1250 mg/kg bw/d. A clear NOAEL of 781 mg/kg bw/d was derived for this study.