Registration Dossier

Administrative data

Description of key information

Oral route:
13 week range finding studies with benzyl alcohol in rats and mice
13 week range finding studies with benzyl acetate in rats and mice
Dermal route:
A 30 day study with benzyl benzoate in rats
A 13 week study with phenyl ethyl alcohol in rats

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Study period:
The 2-year study in rats was begun in November, 1978. The 2-year study in mice was begun in August, 1978.
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 acetate, a substance which has been demonstrated to be very similar in structure, physical/chemical properties and toxicological profile .
Qualifier:
no guideline followed
Principles of method if other than guideline:
Thirteen-week studies were conducted to evaluate the cumulative toxicity of benzyl acetate and to determine the doses to be used in the 2-year studies. Four-week-old male and female F344/ N rats were obtained from Harlan Industries, observed for 13 days, and assigned by species and sex to cages according to a table of random numbers. The cages were then assigned to dosed and control groups according to another table of random numbers. Rats were housed five per cage in polycarbonate cages covered with spun-bonded polyester filters. Racks and filters were replaced every 2 weeks. Cages and bedding were replaced twice per week. Water, via an automatic watering system, and Wayne Lab Blox@ were available ad libitum. Groups of 10 rats of each sex were administered 0, 62.5, 125, 250, 500, or 1,000 mg/ kg benzyl acetate in corn oil by gavage, 5 days per week for 13 weeks. Animals were checked for mortality and signs of morbidity twice daily. Those animals that were judged moribund were killed and necropsied. Each animal was given a clinical examination weekly, including palpation for tissue masses or swelling. Individual body weight data were collected weekly. On days 92-96, survivors were killed with carbon dioxide. Necropsies were performed on animals that survived to the end of the study and on all animals found dead, unless precluded in whole or in part by autolysis or cannibalization. The following specimens were examined microscopically for controls, for the highest dosage group with at least 60% survivors at the time of the group kill, and for all animals that died before the survivors of the group were killed: gross lesions, tissue masses, abnormal lymph nodes, skin, mandibular lymph nodes, mammary gland, salivary gland, thigh muscle, bone, bone marrow, thymus, trachea, lungs and bronchi, heart, thyroid, parathyroid, esophagus, stomach, duodenum, jejunum, ileum, colon, mesenteric lymph nodes, liver, pancreas, spleen, kidneys, adrenals, urinary bladder, seminal vesicles/ prostate/ testes or ovaries/ uterus, nasal cavity, brain, pituitary, and spinal cord, Tissues were preserved in 10% neutral buffered formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Harlan Industries Indianapolis. I N
- Age at study initiation: 6 weeks
- Weight at study initiation: 84.9-127.1
- Housing: Bedding: Beta-Chips heat treated hardwood chips
Cages: Polycarbonate
Cage filters: Spun-bonded polyester filters
- Diet (e.g. ad libitum): Wayne Lab-Blox Allied Mills. ad libitum
- Water (e.g. ad libitum): Tap water supplied through automatic watering system. ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 40-60
- Air changes (per hr): 15
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: Doses were prepared on a weight-to-volume basis by pipetting the appropriate amount of benzyl acetate into a vessel and adding enough corn oil to give the desired concentration. Solutions were mixed until they were visually homogeneous. Rats received 5 ml/kg. Benzyl acetate/corn oil mixtures were analyzed at Midwest Research Institute and found to be stable at room temperature for at least 7 days. Once prepared, benzyl acetate/corn oil mixtures were stored at 5O°C for no longer than 11 days.


VEHICLE
- Justification for use and choice of vehicle (if other than water): No data
- Concentration in vehicle: Male and female rats: 0, 62.5, 125, 250, 500, 1,000 mg/kg body weight in corn oil
- Amount of vehicle (if gavage): 5 ml/kg for rat
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples of benzyl acetate in corn oil were selected at random and analyzed periodically at Southern Research Institute. Results of these analyses and of reference analyses at Midwest Research Institute indicated that benzyl acetate/ corn oil mixtures were properly formulated.
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
5 days/week
Remarks:
Doses / Concentrations:
62.5, 125, 250, 500, 1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10 animals
Control animals:
yes, concurrent vehicle
Details on study design:
Dose selection rationale: based on 14-day study
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Observed daily for mortality and clinical signs.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly clinical examinations

BODY WEIGHT: Yes
- Time schedule for examinations: at the start and end of dosing

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: Necropsies performed on all animals, following tissues examined histologically in control and highest-dose groups with 60% survivors: brain, pituitary, salivary glands, esophagus, mandibular lymph node, thymus, spleen, heart, thyroid, parathyroid. trachea, lungs and bronchi, stomach, liver, large and small intestines, pancreas, mesenteric lymph node, testicles or ovaries, prostate or uterus, seminal vesicles, mammary gland, skin, bone,bone marrow, thigh muscle, kidneys, urinary bladder, adrenal glands, gross lesions, tissues, masses, nasal cavity and spinal cord.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes, following tissues examined histologically in control and highest-dose groups with 60% survilors: brain, pituitary, salivary glands, esophagus, mandibular lymph node, thymus, spleen, heart, thyroid, parathyroid. trachea, lungs and bronchi, stomach, liver, large and small intestines, pancreas, mesenteric lymph node, testicles or ovaries, prostate or uterus, seminal vesicles, mammary gland, skin, bone,bone marrow, thigh muscle, kidneys, urinary bladder, adrenal glands, gross lesions, tissues, masses, nasal cavity and spinal cord.
Other examinations:
No further data
Statistics:
No data
Clinical signs:
effects observed, treatment-related
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:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
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:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
MORTALITY
Two of ten males and 1/10 females that received 1,000 mg/ kg died on day 86. No other animals died during the study.

CLINICAL SIGNS
The only clinical signs attributed to compound administration were observed in male and female rats receiving 1,000 mg/ kg and in females receiving 500 mg/kg. These signs included trembling, ataxia, and sluggishness.

BODY WEIGHT AND WEIGHT GAIN
Final mean body weight in male rats receiving 1,000 mg/kg was about 12% lower than the control group

GROSS PATHOLOGY
Thickened stomach walls were observed in 2/9 males and 4/ 10 females receiving 1,000 mg/kg.

HISTOPATHOLOGY
No compound-related histopathologic effects were observed.
Dose descriptor:
NOAEL
Effect level:
ca. 500
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Ovrall effects Clinical signs; mortality; body weight
Dose descriptor:
NOAEL
Effect level:
ca. 250
Based on:
test mat.
Sex:
female
Basis for effect level:
other: overall effects clinical signs; mortality; body weight
Critical effects observed:
not specified

Male/Female       Dose       Survival       Final weight relative to control %

Male                     0              10/10              -

62.5           10/10              100.4

125            10/10              101.9

250            10/10              102.4

500            10/10              102.6

1000           8/10                 87.6

Female                   0             10/10                  -

62.5           10/10              104.6

125            10/10                99.9       

250            10/10              101.7       

500            10/10              110.0       

1000           9/10                 94.6

Conclusions:
In a 13 week study female and male rats received daily oral doses of 0, 62.5, 125, 250, 500, or 1000 mg/kg bw benzyl acetate. Based on clinical signs, mortality and gross pathology the NOAEL was considered to be 500 and 250 mg/kg body weight for male and female rats, respectively.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
250 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
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 .
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
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
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
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
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
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
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
Critical effects observed:
not specified
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.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
781 mg/kg bw/day
Study duration:
subacute
Species:
rat

Repeated dose toxicity: dermal - local effects

Link to relevant study records
Reference
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 .
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
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
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
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
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
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
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
Critical effects observed:
not specified
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.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Study duration:
subacute
Species:
rat

Additional information

See justification for non-classification


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
For the oral route, a weight of evidence approach was used. The studies included in this weight of evidence were performed for the National Toxicology Program and were of equivalent value. The study selected provided the lowest NOAEL for studies performed with benzyl acetate.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
This study was performed to GLP and using method similar to current guidelines. It was also performed with benzyl benzoate which is the closest analogue to 3-phenyl propyl benzoate (structure, physical/chemical properties and toxicokinetic profile) .

Justification for classification or non-classification

The toxicity of the category members has been investigated in mice and rats by the oral or the dermal routes following subacute (28 days) or subchronic (90 days) exposure. Toxicokinetics studies have shown that they are absorbed via the oral and dermal routes although not at the same rate. The category compounds had an effect on survival, bodyweight and sometimes food consumption at the highest doses. This effect was generally accompanied by systemic toxicity targeting the CNS, kidney, liver and testes. The effects on GIT and skin were due to local toxic effects. Although toxic effects were observed in repeated dose studies performed with the category members, classification as specific target organ toxicity - repeated exposure (STOT – RE) is notrequired since the doses/concentrations which produced the effects are not within the guidance value rangesas indicated in Regulation (EC) No 1272/2008.Based on the results and non-classification of the category members, 3-PPB is not expected to be classified as STOT – RE.