Benzyl acetate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

No specific studies of reproductive toxicity are available for the substance benzyl acetate; however the available evidence (repeated dose toxicity studies with the substance and read-across reproductive toxicity data) clearly indicates a lack of reproductive toxicity potential for this substance. Specific testing for reproductive toxicity in a screening study or in a multi-generation study is not considered to be justified, both scientifically and for reasons of animal welfare.  A number of high quality repeated dose toxicity studies (US NTP) using gavage and dietary administration, performed in the rat and mouse are available for benzyl acetate.  The NTP studies include histopathological evaluation of reproductive tract tissues and a specific assessment of reproductive toxicity potential by investigation of sperm morphology and vaginal cytology in the 13-week dietary studies. Data are also available from a lifetime mouse study with 2% sodium benzoate in drinking water. This study included gross and pathological assessment of the testes and ovaries. A 4 -generation rat study with benzoic acid at concentrations of up to 1% in the diet showed no reproductive effects.

Link to relevant study records

Reference

Reference 1

Endpoint:

reproductive toxicity, other

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1984

Principles of method if other than guideline:

Assessment of toxicity by dosing mice with 2% solution of sodium benzoate in drinking water.

GLP compliance:

no

Limit test:

no

Specific details on test material used for the study:

Sodium benzoate
Molecular weight: 144.11
Melting point: >300 degrees C
Purity: 99%
Source: Fisher Scientifc Co

Species:

mouse

Strain:

Swiss

Sex:

male/female

Details on test animals or test system and environmental conditions:

Source: In-house
Housing: Plastic cages with granular cellulose bedding housed in groups of 10
Diet: Wayne Lab Blox diet
Water: Tap water; ad libitum

Route of administration:

oral: drinking water

Details on exposure:

Sodium benzoate was dissolved in the drinking water as a 2% solution.

A toxicity range-finding study was conducted for 35 days with dosing at 0.5, 1, 2, 4 and 8 % in drinking water. Each group comprised four males and four females. Survival rate, body weight, chemical consumption and histological chnages were assessed. The 2% dose was found to be suitable for longer term exposure based on complete mortality at 8% and some mortality and reduced body weight at 4%.

Details on mating procedure:

Not applicable - study designed to assess the chronic toxiicity of the test material which inlcuded assessment of the ovaries and testes.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Animals dosed for the lifespan of the mice from 39 days old (from weaning).

Frequency of treatment:

Daily

Dose / conc.:

2 other: % in drinking water

No. of animals per sex per dose:

50/sex for treatment group
100/sex for control

Control animals:

yes, concurrent no treatment

Parental animals: Observations and examinations:

Animals were checked and weighed at weekly intervals. Gross pathological changes were recorded. The animals were either allowed to die or were sacrificed using ether when moribund. Complete necropsies were performed on all animals.

Postmortem examinations (parental animals):

All organs were examined macroscopically and fixed in 10% buffered formalin. Histopathological analysis was conducted on the liver, spleen, kidneys, bladder, thyroid, heart, pancreas, testes, ovaries, brain, nasal turbinates, at least four lobes of the lungs and on those organs with pathological chnages. Sections from these tissues were stained with hematoxylin and eosin and examined by light microscopy

Clinical signs:

not specified

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

Treatment had no effect on the survivals when compared with the control group.

Body weight and weight changes:

not specified

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 examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Tumour distribution in control and treated mice

	Untreated controls		2% sodium benzoate in water
	Male	Female	Male	Female
Effective number of mice	99	99	50	50
Number of lung tumours	23	21	7	10
Number of malignant lymphomas	12	24	3	10
Number of blood vessel tumours	6	5	4	2
Number of tumours of testes	0	-	0	-
Number of tumours of the ovary	-	3	-	0
Number of tumours of the uterus	-	1	-	0

 

Executive summary:

Administration of sodoum benzoate at 2% in drinking water daily for life showed no apparent carcinogenic effects. The study included assessment of the testes and ovaries.

Reference 2

Endpoint:

reproductive toxicity, other

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1984

Reason / purpose for cross-reference:

read-across source

Principles of method if other than guideline:

Assessment of toxicity by dosing mice with 2% solution of sodium benzoate in drinking water.

GLP compliance:

no

Limit test:

no

Species:

mouse

Strain:

Swiss

Sex:

male/female

Details on test animals or test system and environmental conditions:

Source: In-house
Housing: Plastic cages with granular cellulose bedding housed in groups of 10
Diet: Wayne Lab Blox diet
Water: Tap water; ad libitum

Route of administration:

oral: drinking water

Details on exposure:

Sodium benzoate was dissolved in the drinking water as a 2% solution.

A toxicity range-finding study was conducted for 35 days with dosing at 0.5, 1, 2, 4 and 8 % in drinking water. Each group comprised four males and four females. Survival rate, body weight, chemical consumption and histological chnages were assessed. The 2% dose was found to be suitable for longer term exposure based on complete mortality at 8% and some mortality and reduced body weight at 4%.

Details on mating procedure:

Not applicable - study designed to assess the chronic toxiicity of the test material which inlcuded assessment of the ovaries and testes.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Animals dosed for the lifespan of the mice from 39 days old (from weaning).

Frequency of treatment:

Daily

Dose / conc.:

2 other: % in drinking water

No. of animals per sex per dose:

50/sex for treatment group
100/sex for control

Control animals:

yes, concurrent no treatment

Parental animals: Observations and examinations:

Animals were checked and weighed at weekly intervals. Gross pathological changes were recorded. The animals were either allowed to die or were sacrificed using ether when moribund. Complete necropsies were performed on all animals.

Postmortem examinations (parental animals):

All organs were examined macroscopically and fixed in 10% buffered formalin. Histopathological analysis was conducted on the liver, spleen, kidneys, bladder, thyroid, heart, pancreas, testes, ovaries, brain, nasal turbinates, at least four lobes of the lungs and on those organs with pathological chnages. Sections from these tissues were stained with hematoxylin and eosin and examined by light microscopy

Clinical signs:

not specified

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

Treatment had no effect on the survivals when compared with the control group.

Body weight and weight changes:

not specified

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 examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Tumour distribution in control and treated mice

	Untreated controls		2% sodium benzoate in water
	Male	Female	Male	Female
Effective number of mice	99	99	50	50
Number of lung tumours	23	21	7	10
Number of malignant lymphomas	12	24	3	10
Number of blood vessel tumours	6	5	4	2
Number of tumours of testes	0	-	0	-
Number of tumours of the ovary	-	3	-	0
Number of tumours of the uterus	-	1	-	0

 

Executive summary:

Administration of sodoum benzoate at 2% in drinking water daily for life showed no apparent carcinogenic effects. The study included assessment of the testes and ovaries.

Reference 3

Endpoint:

multi-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1960

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Principles of method if other than guideline:

A feeding study was performed, in which 4 generations of rats received the test substance (0.5% or 1%). The first generation was exposed for 8 weeks and then allowed to mate (1/1 for a period of 14 days). Mating was repeated in week 48 to raise a second litter. Survival of the first and second generation was measured. The third generation was terminated after 16 weeks and examined histopathologically. In this generation weights of brain, heart, liver, spleen, kidneys and testes were determined. The fourth generation was terminated after weaning of the pups. Body weights were determined in week 4, 8 and 12 weeks of each generation (week 12 males only). Feeding efficiency was measured in all generations after 2,4, 6 and 8 weeks. Some reproduction parameters were assessed: percentage of infertility, sexual maturation, litter size, total pups and surviving pups. These parameters were assessed for all generations (summed) and for the first two generations separately.

GLP compliance:

no

Remarks:

Study conducted prior to GLP

Limit test:

no

Specific details on test material used for the study:

Benzoic acid

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

Source: Farbwerken Bayer
Weight at study initiation: 40-50g
Housing: Double cages
Diet: paired feeding for first 8 weeks, then ad libitum
Water: ad libitum

Route of administration:

oral: feed

Vehicle:

other: Mixed with diet

Details on exposure:

Feed mixtures were produced in a feed mixing machine made of stainless steel. Diet consisted of commercial standard rat feed made by Lutz (Euskirchen) with sufficient benzoic acid added to achieve feed concentrations of 0.5% and 1.0%.

Details on mating procedure:

1 male and 1 female cohabited for 14 days. If unsuccessful mating, this was repeated 8 weeks later to investigate delays in sexual maturity or sterility.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

No futher details available

Duration of treatment / exposure:

11-12 weeks prior to mating and through 4 generations.

Frequency of treatment:

Feed available ad libitum

Details on study schedule:

No futher details available

Dose / conc.:

0 other: % in diet

Remarks:

Untreated control

Dose / conc.:

0.5 other: % in diet

Remarks:

Equivalent to 450 and 600 mg/kg bw/day for males and females, respectively.

Dose / conc.:

1 other: % in diet

Remarks:

Equivalent to 900 and 1116 mg/kg bw/day for males and females, respectively.

No. of animals per sex per dose:

20/sex/group/generation

Control animals:

yes, plain diet

Details on study design:

The dietary concentrations were selected based upon a previous range-finding study where rats consuming 5% benzoic acid in the diet died within 3 weeks due to lack of palatability of the diet and corrosive effects of the free acid on the digestive tract.

Positive control:

None stated

Parental animals: Observations and examinations:

Body weight recorded weekly for the first 8 weeks, then every 4 weeks.
Food consumption for each animal recorded. Consumption data presented as 'protein efficiency (weight increase per gram of dietary protein)'. Compound intake calculated as time-weighted averages from the consumption and body weight gain.

Oestrous cyclicity (parental animals):

Not examined

Sperm parameters (parental animals):

Testes weight examined in the third generation

Litter observations:

Litter size recorded on first and second days. Surviving young determined on twenty-first day.
As there were no differences between generations, data were combined from the four generations.
Assessments included total number of pups born, pup survival and litter size.

Postmortem examinations (parental animals):

The third generation was subject to a necropsy after 16 weeks of exposure. Organ weights (brain, heart, spleen, liver, kidneys and testes) were taken. Organs were examined histopathologically.

Postmortem examinations (offspring):

None stated

Statistics:

Not provided

Reproductive indices:

Not provided

Offspring viability indices:

No differences noted between generations; all data were combined.

Clinical signs:

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

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

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Histopathology was only reported for 3rd generation animals.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group. There were no differences in reproduction or development of the young in the 4 observed generations exposed to 1% benzoic acid in the diet.

Key result

Dose descriptor:

NOAEL

Effect level:

1 other: %

Based on:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

Generational data were not reported separately. At concentrations up to 1% benzoic acid in the diet, there were no adverse effects on parents or offspring through four generations of test substance administration. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Clinical signs:

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption (efficiency) over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Organ weights were only reported for the 3rd generation animals.

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Histopathology was only reported for 3rd generation animals.

Histopathological findings: neoplastic:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group.There were no differences in reproduction or development of the young in the 4 observed generations exposed to 1% benzoic acid in the diet.

Key result

Dose descriptor:

NOEL

Effect level:

> 1 other: %

Based on:

other:

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

Generational data were not reported separately. At concentrations up to 1% benzoic acid in the diet, there were no adverse effects on parents or offspring through four generations of test substance administration. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Clinical signs:

not specified

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption (efficiency) over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

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

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Organ weights were only reported for the 3rd generation animals

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Description (incidence and severity):

Histopathology was only reported for the 3rd generation animals.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group.

Key result

Dose descriptor:

NOEL

Generation:

F1

Effect level:

> 1 other: %

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

There were no adverse effects on viability, sexual maturation, mortality, body weight and weight gain, food consumption and efficiency, body weights, select organ weight and pathology for any of the four generations in this study receiving up to 1% benzoic acid in the diet. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Clinical signs:

not specified

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption (efficiency) over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Organ weights were only reported for the 3rd generation animals.

Gross pathological findings:

not specified

Histopathological findings:

no effects observed

Description (incidence and severity):

Histopathology was only reported for 3rd generation animals.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group.

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

1 other: %

Based on:

other:

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

There were no adverse effects on viability, sexual maturation, mortality, body weight and weight gain, food consumption and efficiency, body weights, select organ weight and pathology for any of the four generations in this study receiving up to 1% benzoic acid in the diet. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Conclusions:

There were no adverse effects on reproductive parameters including fertility measures, delayed sexual maturity, total number of pups born, pup survival, onset of reproductive senescence or litter size when male and female rats were fed up to 1% benzoic acid in the diet over four generations. Under conditions of this study, benzoic acid is not a reproductive toxicant.

Executive summary:

In a long-term feeding study, benzoic acid was added to standard feed to achieve concentration of 0.5% or 1.0% in the diet. Untreated controls were also included. Diets were provided ad libitum to groups of 20 rats/sex through four generations. Body weights and feed consumption data were collected throughout the exposure period. Other endpoints examined included: onset of sexual maturity, evidence of permanent sterility, onset of menopause, litter sizes, number of pups born, surviving young, organ weights and histology, and effect on lifespan. Feed consumption, body weights, and weight gain were unaffected by exposure to benzoic acid at concentrations up to 1% in the diet. There was actually an unexplained statistically significant increase in the lifespan of rats in the 0.5% exposure group, i.e., a higher percentage of rats lived longer. There were no differences between the groups exposed to benzoic acid and the control group for fertility measures, delayed sexual maturity, total number of pups born, pup survival, onset of reproductive senescence or litter size. In addition, organ weights and histopathologic findings were similar for all groups. Under conditions of this study, there were no dose-related adverse effects on either reproductive or developmental parameters in both sexes of rats fed 1% benzoic acid in the diet over four generations.

Reference 4

Endpoint:

multi-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1960

Reliability:

2 (reliable with restrictions)

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Principles of method if other than guideline:

A feeding study was performed, in which 4 generations of rats received the test substance (0.5% or 1%). The first generation was exposed for 8 weeks and then allowed to mate (1/1 for a period of 14 days). Mating was repeated in week 48 to raise a second litter. Survival of the first and second generation was measured. The third generation was terminated after 16 weeks and examined histopathologically. In this generation weights of brain, heart, liver, spleen, kidneys and testes were determined. The fourth generation was terminated after weaning of the pups. Body weights were determined in week 4, 8 and 12 weeks of each generation (week 12 males only). Feeding efficiency was measured in all generations after 2,4, 6 and 8 weeks. Some reproduction parameters were assessed: percentage of infertility, delayed sexual maturation, litter size, total pups and surviving pups. These parameters were assessed for all generations (summed) and for the first two generations separately.

GLP compliance:

no

Remarks:

Study conducted prior to GLP

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

Source: Farbwerken Bayer
Weight at study initiation: 40-50g
Housing: Double cages
Diet: paired feeding for first 8 weeks, then ad libitum
Water: ad libitum

Route of administration:

oral: feed

Vehicle:

other: Mixed with diet

Details on exposure:

Feed mixtures were produced in a feed mixing machine made of stainless steel. Diet consisted of commercial standard rat feed made by Lutz (Euskirchen) with sufficient benzoic acid added to achieve feed concentrations of 0.5% and 1.0%.

Details on mating procedure:

1 male and 1 female cohabited for 14 days. If unsuccessful mating, this was repeated 8 weeks later to investigate delays in sexual maturity or sterility.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

No futher details available

Duration of treatment / exposure:

11-12 weeks prior to mating and through 4 generations.

Frequency of treatment:

Feed available ad libitum

Details on study schedule:

No futher details available

Dose / conc.:

0.5 other: % in diet

Remarks:

Equivalent to 450 and 600 mg/kg bw/day for males and females, respectively.

Dose / conc.:

1 other: % in diet

Remarks:

Equivalent to 900 and 116 mg/kg bw/day for males and females, respectively.

No. of animals per sex per dose:

20/sex/group/generation

Control animals:

yes, plain diet

Details on study design:

The dietary concentrations were selected based upon a previous range-finding study where rats consuming 5% benzoic acid in the diet died within 3 weeks due to lack of palatability of the diet and corrosive effects of the free acid on the digestive tract.

Positive control:

None stated

Parental animals: Observations and examinations:

Body weight recorded weekly for the first 8 weeks, then every 4 weeks.
Food consumption for each animal recorded. Consumption data presented as 'protein efficiency (weight increase per gram of dietary protein)'. Compound intake calculated as time-weighted averages from the consumption and body weight gain.

Oestrous cyclicity (parental animals):

Not examined

Sperm parameters (parental animals):

Testes weight examined in the third generation

Litter observations:

Litter size recorded on first and second days. Surviving young determined on twenty-first day.
As there were no differences between generations, data were combined from the four generations.
Assessments included total number of pups born, pup survival and litter size.

Postmortem examinations (parental animals):

The third generation was subject to a necropsy after 16 weeks of exposure. Organ weights (brain, heart, spleen, liver, kidneys and testes) were taken. Organs were examined histopathologically.

Postmortem examinations (offspring):

None stated

Statistics:

Not provided

Reproductive indices:

Not provided

Offspring viability indices:

No differences noted between generations; all data were combined.

Clinical signs:

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption (efficiency) over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Histopathology was only reported for 3rd generation animals.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group. There were no differences in reproduction or development of the young in the 4 observed generations exposed to 1% benzoic acid in the diet. See Table 1 in "Any other information on results incl. tables".

Key result

Dose descriptor:

NOEL

Effect level:

> 1 other: %

Based on:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

Generational data were not reported separately. At concentrations up to 1% benzoic acid in the diet, there were no adverse effects on parents or offspring through four generations of test substance administration. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Clinical signs:

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption (efficiency) over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Organ weights were only reported for the 3rd generation animals.

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Histopathology was only reported for 3rd generation animals.

Histopathological findings: neoplastic:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group.There were no differences in reproduction or development of the young in the 4 observed generations exposed to 1% benzoic acid in the diet. See Table 1 in "Any other information on results incl. tables".

Key result

Dose descriptor:

NOEL

Effect level:

> 1 other: %

Based on:

other:

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

Generational data were not reported separately. At concentrations up to 1% benzoic acid in the diet, there were no adverse effects on parents or offspring through four generations of test substance administration. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Clinical signs:

not specified

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption (efficiency) over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Organ weights were only reported for the 3rd generation animals

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Description (incidence and severity):

Histopathology was only reported for the 3rd generation animals.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group.

Key result

Dose descriptor:

NOEL

Generation:

F1

Effect level:

> 1 other: %

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

There were no adverse effects on viability, sexual maturation, mortality, body weight and weight gain, food consumption and efficiency, body weights, select organ weight and pathology for any of the four generations in this study receiving up to 1% benzoic acid in the diet. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Clinical signs:

not specified

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

There was no difference between the 1% group and the control group in terms of lifespan (similar number of short vs long-lived animals) but there was a statistically significant increase in the number of long-lived animals in the 0.5% group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on body weight gain or body weights over the four generations of rats tested in this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There was no effect of feeding 0.5 or 1.0% benzoic acid in the diet on feed consumption (efficiency) over the four generations of rats tested in this study.

Food efficiency:

no effects observed

Description (incidence and severity):

Feed consumption data in this study are presented as “Protein efficiency (weight increase per gram of dietary protein)”. There was no effect on feed consumption (efficiency) in either test group.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Organ weights were only reported for the 3rd generation animals.

Gross pathological findings:

not specified

Histopathological findings:

no effects observed

Description (incidence and severity):

Histopathology was only reported for 3rd generation animals.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

For all four generations, there were no adverse effects on food consumption or efficiency, body weights, life span, organ weights, or organ pathology in male and female rats receiving 1% benzoic acid in the diet when compared to the control group.

Key result

Dose descriptor:

NOEL

Generation:

F2

Effect level:

> 1 other: %

Based on:

other:

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

There were no adverse effects on viability, sexual maturation, mortality, body weight and weight gain, food consumption and efficiency, body weights, select organ weight and pathology for any of the four generations in this study receiving up to 1% benzoic acid in the diet. Overall, the dose level from the 1% diet for the entire premating period was approximately 900 and 1176 mg/kg/day for the male and female rats, respectively.

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Conclusions:

In a chronic feeding study, there were no adverse effects on reproductive parameters including fertility measures, delayed sexual maturity, total number of pups born, pup survival, onset of reproductive senescence or litter size when male and female rats were fed up to 1% benzoic acid in the diet over four generations. Under conditions of this study, benzoic acid is not a reproductive toxicant.

Executive summary:

In a long-term feeding study, benzoic acid was added to standard feed to achieve concentration of 0.5% or 1.0% in the diet. Untreated controls were also included. Diets were provided ad libitum to groups of 20 rats/sex through four generations. Body weights and feed consumption data were collected throughout the exposure period. Other endpoints examined included: onset of sexual maturity, evidence of permanent sterility, onset of menopause, litter sizes, number of pups born, surviving young, organ weights and histology, and effect on lifespan. Feed consumption, body weights, and weight gain were unaffected by exposure to benzoic acid at concentrations up to 1% in the diet. There was actually an unexplained statistically significant increase in the lifespan of rats in the 0.5% exposure group, i.e., a higher percentage of rats lived longer. There were no differences between the groups exposed to benzoic acid and the control group for fertility measures, delayed sexual maturity, total number of pups born, pup survival, onset of reproductive senescence or litter size. In addition, organ weights and histopathologic findings were similar for all groups. Under conditions of this study, there were no dose-related adverse effects on either reproductive or developmental parameters in both sexes of rats fed 1% benzoic acid in the diet over four generations.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Data for benzyl acetate

 

NTP gavage studies

 

Fourteen-day range-finding gavage studies in the rat (using dose levels of 0, 250, 500, 1000, 2000 or 4000 mg/kg bw/d) and the mouse (using dose levels of 0, 125, 250, 500, 1000 or 2000 mg/kg bw/d) did not show any gross evidence of any effects of treatment on the reproductive system at dose levels causing mortality. It is recognised that no histopathological investigations were performed in these studies, thereby limiting their sensitivity.

 

Thirteen-week studies performed in the rat (using dose levels of 0, 62.5, 15, 250, 500 or 1000 mg/kg bw/d) and mouse (using dose levels of 0, 125, 250, 500, 1000 or 2000 mg/kg bw/d) included histopathological investigation of theseminal vesicles, prostate, testes, ovaries and uterus. The high dose levels in these studies were sufficient to cause mortality. In the rat study, treatment-related effects at gross necropsy were limited to thickened stomach walls in both sexes at 1000 mg/kg bw/d. No treatment-related histopathological effects were observed at any dose level. In the mouse study, no treatment-related gross or microscopic effects were observed at any dose level.

 

Two-year NTP carcinogenicity studies were also performed with benzyl acetate in rats (using gavage dose levels of 0, 250 or 500 mg/kg bw/d) and in mice (using gavage dose levels of 0, 500 or 1000 mg/kg bw/d). The studiesincluded histopathological investigation of theseminal vesicles, prostate, testes, ovaries and uterus. No treatment-related effects on the reproductive system were noted in either species.

 

NTP dietary studies

 

The 13-week dietary studies included gross necropsy of all animals; weights of the prostate, seminal vesicle, testis and uterus (rat) were recorded. Histopathological investigations included assessment of the testes, epididymides, seminal vesicles, ovaries and uterus.

 

The 13-week rat study was performed at dose levels of 0, 3130, 6250, 12500, 25000 or 50000 ppm benzyl acetate. Dietary dose levels were equivalent to mean intakes of 0, 230, 460, 900, 1750 and 3900 mg/kg bw/d in males; 0, 240, 480, 930, 1870 and 4500 mg/kg bw/d in females. Overt toxicity was seen, with high (90%) mortality at the top dose level of 50000 ppm, changes in bodyweight and/or food consumption at 12500, 25000 and 50000 ppm.Testicular changes were seen in male rats at 12500, 25000 and 50000 ppm. Findings were characterised by mild or moderate aspermatogenesis in two rats at 50000 ppm, with atrophy of the seminiferous tubules in two rats at 25000 ppm and one rat at 12500 ppm rats. In one rat at 25000 ppm, seminiferous tubular atrophy was marked in severity and included atypical cells (enlarged, degenerated spermatozoa) in the corresponding epididymis and mild interstitial cell hyperplasia. The seminiferous tubule atrophy was minimal in severity in the single rat affected at 12500 ppm. No testicular findings were observed at dose levels of 6250 ppm or lower exposure levels. Additional findings in rats at 50000 ppm included secretory depletion of the seminal vesicles, and immature, hypoplastic uterus. However it is noted that these changes are frequently observed in rodents and are usually secondary effects of general toxicity. No treatment-related effects in any organ were noted in the two-year study, which was performed at dose levels of 0, 3000, 6000 and 12000 ppm.

 

The 13-week mouse study was performed at dose levels of 0, 3130, 6250, 12500, 25000 or 50000 ppm benzyl acetate. Dietary dose levels were equivalent to mean intakes of 0, 425, 1000, 2000m 3700 and 7900 mg/kg bw/d in males; 0, 650, 1280, 2980, 4300 and 9400 mg/kg bw/d in females.No relevant findings were noted on the tissues of the reproductive tract in either the 13-week study or in the two year study at dose levels of 0, 330, 1000 and 3000 ppm.

 

Additional specific investigations of reproductive function were made in both species at the end of the 13-week dietary studies. Sperm morphology and vaginal cytology of rats and mice from all dose levels were evaluated. No treatment-related effects on sperm morphology occurred in rats or mice.Asignificant dose-related of the oestrous cycle occurred in female mice; this effect did not occur in female rats. The lengthening of the oestrous cycle observed in exposed female mice is considered to be related to the reduced body weights seen in this study and is not a direct toxic effect of the test material.

 

The data from the rat 13-week dietary study with benzyl acetate indicate a potential effect on the male reproductive tract. With the exception of a single male at 12500 ppm (with findings described as ‘minimal’), findings were associated with toxicity (including 90% mortality at the top dose level) and bodyweight effects and may therefore be secondary, rather than direct toxic effects of the test material. The single, minimal grade incidence at 12500 ppm is not considered to be clearly related to treatment. It is notable that similar findings were not seen in the gavage study, which used comparable dose levels and in which the bolus dosing would be expected to produce more severe effects. The absence of effects in the rat following gavage or dietary administration for two years is equally notable, and specific investigations at the end of the 13-week dietary study did not reveal any effects on sperm morphology.

 

Read-across data

 

Benzyl acetate is rapidly hydrolysed by plasma and tissue esterases to benzoic acid, with little or no systemic exposure to the intact substance. The related compounds benzaldehdye and benzyl alcohol are similarly metabolised to benzoic acid. Read-across from studies using benzoic acid, benzaldehdye or benzyl alcohol is therefore relevant for benzyl acetate. This approach to substance grouping, based on common metabolism to benzoic acid, has also been taken by a number of international bodies including the WHO in its consideration of the use of benzyl acetate as a food additive (WHO Food Additives Series 37), the SCCP in its consideration of the use of benzoic acid in consumer products (SCCP/0891/050) the OECD (SIDS Initial Assessment Report for 13^thSIAM, 2001) and IPCS (CICAD 26).  

 

The WHO (Food Additives Series 37) Committee concluded that the data reviewed on the substances in this group (including the NTP studies) were sufficient to demonstrate a lack of reproductive toxicity potential.

 

The SCCP evaluation of benzoic acid (SCCP/0891/05) notes the results of a 4-generation reproductive toxicity study (Kieckebusch and Lang, 1960) in rats at dietary dose levels of up to1%. No adverse effects were noted in this study and a NOAEL equivalent to 500 mg/kg bw/d was determined for this study.

The data reported by Toth (1984) showed no evidence of carcinogenicity in mice for the lifetime of the animals following dosing with sodium benzoate at 2% in drinking water (equivalent to 6200 mg/kg bw/day). The study included assessement of the testes and ovaries.

 

The IPCS evaluation of benzoic acid includes an assessment of the available data for benzoic acid and related substances (including benzyl acetate) and concludes that reproductive toxicity is unlikely at dose levels not causing maternal toxicity.

 

The OECD SIDS for the benzoate group of compounds including benzoic acid and benzyl alcohol (SIDS Initial Assessment Report for 13^thSIAM, 2001) evaluated the data available for this group of compounds (including benzyl acetate) and concluded that the absence oftreatment-related effects on reproductive organs in the (sub)chronic studies with all compounds supports the lack of reproductive potential.

 

Conclusion

 

The high quality repeated dose toxicity NTP studies for benzyl acetate do not show any clear effects of the substance on the reproductive tract of males or females and no effects on sperm morphology or vaginal cytology. Findings therefore do not indicate a potential for reproductive toxicity. Read-across data for related compounds including a 4-generation study performed with benzoic acid and a lifetime dosing study in mice with sodium benzoate have

been reviewed by a number of international bodies, who conclude that the group of related substances including benzyl acetate is without reproductive toxicity potential.

 

Additional, specific testing for the reproductive toxicity of benzyl acetate in a screening study or in a multi-generation study is therefore not considered to be justified, both scientifically and for reasons of animal welfare.

 

Short description of key information:
No specific studies of reproductive toxicity are available for the substance benzyl acetate; however the available evidence (repeated dose toxicity studies with the substance and read-across reproductive toxicity data with sodium benzoate and benzoic acid) clearly indicates a lack of reproductive toxicity potential for this substance. Specific testing for reproductive toxicity in a screening study or in a multi-generation study with benzyl acetate is considered not to be justified, both scientifically and for reasons of animal welfare.

In addition, a number of high quality repeated dose toxicity studies (US NTP) using gavage and dietary administration, performed in the rat and mouse are available for benzyl acetate. The NTP studies include histopathological evaluation of reproductive tract tissues and a specific assessment of reproductive toxicity potential by investigation of sperm morphology and vaginal cytology in the 13-week dietary studies.

Effects on developmental toxicity

Description of key information

Two study report on developmental toxicity/teratogenicity are available. One study was conducted using benzyl acetate with Wistar rats as the test species. There was no indication it was conducted according to any guidelines or to GLP. The second study reported was conducted using sodium benzoate with Dutch-belted rabbits as the test species.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1972

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Principles of method if other than guideline:

Females artifically inseminated with 0.3 mL of diluted semen from a proven donor buck using approximatley 20 x 10^6 motile sperm according to the procedure described by Vogin et al (Pharmacologist 11, 282 (1969). The females were dosed by oral gavage on Day 6 to 18 of gestation. Body weights were recorded on Days 0, 6, 12, 18 and 29 of gestation. All animals were observed daily for appearance and behaviour, including food consumption and body weight. Numbers of corpora lutea, implantation sites and resorption sites were recorded. The numbers of live and dead foetuses were recorded. Body weights of the live pups were recorded. The urogenital tract of each animal was examined in detail for normality. All foetuses underwent a detailed gross examination for the presence of external congenital abnormalities. The live foetuses were placed in an incubator for 24 hours for the evaluation of neonatal survival. All surviving pups were sacrificed and all pups were examined for visceral abnormalities by dissection. All foetuses were then cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.

GLP compliance:

no

Specific details on test material used for the study:

Sodium benzoate

Species:

rabbit

Strain:

other: Dutch-belted

Details on test animals or test system and environmental conditions:

Housed individually in mesh bottom cages
Temperature and humidity controlled
Food and fresh tap water ad libitum

Route of administration:

oral: gavage

Details on exposure:

Females were dosed by oral gavage on Day 6 to 18 of gestation.

Analytical verification of doses or concentrations:

no

Details on mating procedure:

Females artifically inseminated with 0.3 mL of diluted semen from a proven donor buck using approximatley 20 x 10^6 motile sperm according to the procedure described by Vogin et al (Pharmacologist 11, 282 (1969).

Duration of treatment / exposure:

Days 6 to 18 of gestation.

Frequency of treatment:

Daily

Duration of test:

29 days

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Sham exposed controls

Dose / conc.:

2.5 mg/kg bw/day (nominal)

Dose / conc.:

12 mg/kg bw/day (nominal)

Dose / conc.:

54 mg/kg bw/day (nominal)

Dose / conc.:

250 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Sham treatment: 17 females mated
Positive control: 15 females mated
2.5 mg/kg bw/day: 32 females mated
12.0 mg/kg bw/day: 21 females mated
54 mg/kg bw/day: 22 females mated
250 mg.kg bw/day: 14 females mated

Control animals:

yes, sham-exposed

other:

Details on study design:

All does were subjected to Caesarean section under surgical anaesthesia on Day 29 of gestation.

Maternal examinations:

Body weights were recorded on Days 0, 6, 12, 18 and 29 of gestation. All animals were observed daily for appearance and behaviour, including food consumption and body weight.

Ovaries and uterine content:

Numbers of corpora lutea, implantation sites and resorption sites were recorded.

Fetal examinations:

The numbers of live and dead foetuses were recorded. Body weights of the live pups were recorded. The urogenital tract of each animal was examined in detail for normality. All foetuses underwent a detailed gross examination for the presence of external congenital abnormalities. The live foetuses were placed in an incubator for 24 hours for the evaluation of neonatal survival. All surviving pups were sacrificed and all pups were examined for visceral abnormalities by dissection. All foetuses were then cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.

Clinical signs:

effects observed, treatment-related

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

no effects observed

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

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

not examined

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined

Changes in number of pregnant:

no effects observed

Other effects:

no effects observed

Key result

Remarks on result:

not determinable due to adverse toxic effects at highest dose / concentration tested

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

not specified

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

250 mg/kg bw/day

Based on:

test mat.

Sex:

female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

Fate of doe rabbits

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Total mated	17	32	21	22	14	15
Total pregnant	10	12	10	12	10	10
Total surviving at term	16	28	21	20	12	15
Pregnant surviving at term	10	10	10	11	9	10

 

Maternal body weight (g)

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Day 0	2.69	2.25	2.30	2.53	2.62	2.78
Day 6	2.45	2.29	2.37	2.63	2.74	2.86
Day 12	2.82	2.31	2.32	2.63	2.73	-
Day 18	2.87	2.37	2.34	2.60	2.76	-
Day 29	2.90 (10)	2.47 (10)	2.38 (10)	2.64 (11)	2.86 (9)	2.95 (10)

Number of surviving dams in parenthesis

Litter data

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Total pregnancies	10	12	10	12	10	10
Died or aborted before Day 29	1	4	0	2	2	0
To term pregnancies (on Day 29)	10	10	10	11	9	10
Total corpora lutea	170	302	204	217	207	185
Mean corpora lutea/dam mated	10.0	9.44	9.71	9.86	14.8	12.3
Number of live litters	9	10	8	5	8	7
Total implant sites	59	54	55	51	65	71
Mean implant sites/dam	5.90	5.40	5.50	4.64	7.22	7.10
Total number of resorptions	6	14	13	36	12	39
Dams with 1 or more sites resorbed	3	6	4	11	5	9
Dams with all sites resorbed	1	-	1	6	1	3
Percent partial resorption	30.0	60.0	40.0	100.0	55.6	90.0
Percent complete resorption	10.0	-	10.0	54.6	11.1	30.0
Live foetuses	52	39	36	15	53	30
Average/dam	5.20	3.90	3.60	1.36	5.89	3.00
Sex ratio (M/F)	0.73	1.44	0.89	0.50	0.77	0.85
Dead foetuses	1	1	6	-	-	2
Dams with 1 or more dead foetus	1	1	1	-	-	2
Dams with all dead foetuses	-	-	1	-	-	-
Percent partial dead foetuses	10.0	10.0	10.0	-	-	20.0
Percent all dead foetuses	-	-	10.0	-	-	-
Average foetus weight (g)	38.0	35.4	36.3	41.9	38.6	32.4

Skeletal findings

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Live foetuses examined at term	50/9	39/10	36/8	15/5	53/8	26/7
Incomplete oscillation of sternebrae		1/1	1/1			6/4
Bipartite sternebrae			1/1			1/1
Fused sternebrae		2/2		1/1		8/4
Extra sternebrae	2/2	2/2	1/1		3/2	2/1
Fused or split ribs						7/4
Scrambled vertebrae						10/5
Scoliosis of vertebrae						3/2
Tail defects						26/7
Craniostosis			1/1			2/1

Data presented as number of foetuses affected/number of litters affected

Conclusions:

No evidence of maternal or developmental toxicity was seen in this study.

Executive summary:

The administration of up to 250 mg/kg bw/d of sodium benzoate to pregnant rabbits for 13 consecutive days (from Day 6 to 18 of gestation) had no clearly discernible effect on nidation or on maternal or foetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham-treated controls.

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1972

Reliability:

2 (reliable with restrictions)

Reason / purpose for cross-reference:

read-across source

Qualifier:

no guideline followed

Version / remarks:

Study predates regulatory guidelines

Principles of method if other than guideline:

Females artifically inseminated with 0.3 mL of diluted semen from a proven donor buck using approximatley 20 x 10^6 motile sperm according to the procedure described by Vogin et al (Pharmacologist 11, 282 (1969). The females were dosed by oral gavage on Day 6 to 18 of gestation. Body weights were recorded on Days 0, 6, 12, 18 and 29 of gestation. All animals were observed daily for appearance and behaviour, including food consumption and body weight.
Numbers of corpora lutea, implantation sites and resorption sites were recorded.
The numbers of live and dead foetuses were recorded. Body weights of the live pups were recorded. The urogenital tract of each animal was examined in detail for normality. All foetuses underwent a detailed gross examination for the presence of external congenital abnormalities. The live foetuses were placed in an incubator for 24 hours for the evaluation of neonatal survival. All surviving pups were sacrificed and all pups were examined for visceral abnormalities by dissection. All foetuses were then cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.

GLP compliance:

no

Species:

rabbit

Strain:

other: Dutch-belted

Details on test animals or test system and environmental conditions:

Housed individually in mesh bottom cages
Temperature and humidity controlled
Food and fresh tap water ad libitum

Route of administration:

oral: gavage

Details on exposure:

Females were dosed by oral gavage on Day 6 to 18 of gestation.

Analytical verification of doses or concentrations:

no

Details on mating procedure:

Females artifically inseminated with 0.3 mL of diluted semen from a proven donor buck using approximatley 20 x 10^6 motile sperm according to the procedure described by Vogin et al (Pharmacologist 11, 282 (1969).

Duration of treatment / exposure:

Days 6 to 18 of gestation.

Frequency of treatment:

Daily

Duration of test:

29 days

Dose / conc.:

2.5 mg/kg bw/day (nominal)

Dose / conc.:

12 mg/kg bw/day (nominal)

Dose / conc.:

54 mg/kg bw/day (nominal)

Dose / conc.:

250 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Sham treatment: 17 females mated
Positive control: 15 females mated
2.5 mg/kg bw/day: 32 females mated
12.0 mg/kg bw/day: 21 females mated
54 mg/kg bw/day: 22 females mated
250 mg.kg bw/day: 14 females mated

Control animals:

yes, sham-exposed

other:

Details on study design:

All does were subjected to Caesarean section under surgical anaesthesia on Day 29 of gestation.

Maternal examinations:

Body weights were recorded on Days 0, 6, 12, 18 and 29 of gestation. All animals were observed daily for appearance and behaviour, including food consumption and body weight.

Ovaries and uterine content:

Numbers of corpora lutea, implantation sites and resorption sites were recorded.

Fetal examinations:

The numbers of live and dead foetuses were recorded. Body weights of the live pups were recorded. The urogenital tract of each animal was examined in detail for normality. All foetuses underwent a detailed gross examination for the presence of external congenital abnormalities. The live foetuses were placed in an incubator for 24 hours for the evaluation of neonatal survival. All surviving pups were sacrificed and all pups were examined for visceral abnormalities by dissection. All foetuses were then cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.

Clinical signs:

effects observed, treatment-related

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

no effects observed

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

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

not examined

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined

Changes in number of pregnant:

no effects observed

Other effects:

no effects observed

Key result

Remarks on result:

not determinable due to adverse toxic effects at highest dose / concentration tested

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

not specified

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Other effects:

not specified

Key result

Remarks on result:

not determinable due to adverse toxic effects at highest dose / concentration tested

Developmental effects observed:

no

Fate of doe rabbits

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Total mated	17	32	21	22	14	15
Total pregnant	10	12	10	12	10	10
Total surviving at term	16	28	21	20	12	15
Pregnant surviving at term	10	10	10	11	9	10

 

Maternal body weight (g)

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Day 0	2.69	2.25	2.30	2.53	2.62	2.78
Day 6	2.45	2.29	2.37	2.63	2.74	2.86
Day 12	2.82	2.31	2.32	2.63	2.73	-
Day 18	2.87	2.37	2.34	2.60	2.76	-
Day 29	2.90 (10)	2.47 (10)	2.38 (10)	2.64 (11)	2.86 (9)	2.95 (10)

Number of surviving dams in parenthesis

Litter data

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Total pregnancies	10	12	10	12	10	10
Died or aborted before Day 29	1	4	0	2	2	0
To term pregnancies (on Day 29)	10	10	10	11	9	10
Total corpora lutea	170	302	204	217	207	185
Mean corpora lutea/dam mated	10.0	9.44	9.71	9.86	14.8	12.3
Number of live litters	9	10	8	5	8	7
Total implant sites	59	54	55	51	65	71
Mean implant sites/dam	5.90	5.40	5.50	4.64	7.22	7.10
Total number of resorptions	6	14	13	36	12	39
Dams with 1 or more sites resorbed	3	6	4	11	5	9
Dams with all sites resorbed	1	-	1	6	1	3
Percent partial resorption	30.0	60.0	40.0	100.0	55.6	90.0
Percent complete resorption	10.0	-	10.0	54.6	11.1	30.0
Live foetuses	52	39	36	15	53	30
Average/dam	5.20	3.90	3.60	1.36	5.89	3.00
Sex ratio (M/F)	0.73	1.44	0.89	0.50	0.77	0.85
Dead foetuses	1	1	6	-	-	2
Dams with 1 or more dead foetus	1	1	1	-	-	2
Dams with all dead foetuses	-	-	1	-	-	-
Percent partial dead foetuses	10.0	10.0	10.0	-	-	20.0
Percent all dead foetuses	-	-	10.0	-	-	-
Average foetus weight (g)	38.0	35.4	36.3	41.9	38.6	32.4

Skeletal findings

	Sham 0 mg/kg bw/day	2.5 mg/kg bw/day	12.0 mg/kg bw/day	54.0 mg/kg bw/day	250 mg/kg bw/day	Positive control
Live foetuses examined at term	50/9	39/10	36/8	15/5	53/8	26/7
Incomplete oscillation of sternebrae		1/1	1/1			6/4
Bipartite sternebrae			1/1			1/1
Fused sternebrae		2/2		1/1		8/4
Extra sternebrae	2/2	2/2	1/1		3/2	2/1
Fused or split ribs						7/4
Scrambled vertebrae						10/5
Scoliosis of vertebrae						3/2
Tail defects						26/7
Craniostosis			1/1			2/1

Data presented as number of foetuses affected/number of litters affected

Executive summary:

The administration of up to 250 mg/kg bw/day of sodium benzoate to pregnant rabbits for 13 consecutive days (from Day 6 to 18 of gestation) had no clearly discernible effect on nidation or on maternal or foetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham-treated controls.

Reference 3

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not documented

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

Test animals were exposed to the test substance from gestation day 6 through 15 at 5 different test concentrations, specifically 0, 10, 100, 500 and 1000 mg/kg. The effects of the test substance on both maternal animals and foetuses were subsequently examined.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): Benzyl Acetate

- Substance type: Not documented
- Physical state: Transparent liquid
- Analytical purity: 99%
- Impurities (identity and concentrations): Not documented
- Composition of test material, percentage of components: Not documented
- Isomers composition: Not documented
- Purity test date: Not documented
- Lot/batch No.: AVO1

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Keari Inc.
- Age at study initiation: Females: 10 to 15 weeks
Males: 12 to 17 weeks.
- Weight at study initiation: Not documented
- Fasting period before study: Not documented
- Housing: Not documented
- Diet (e.g. ad libitum): Solid food purchased form Japan Kurea
- Water (e.g. ad libitum): Tap water
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1°C
- Humidity (%): 50 ± 10%
- Air changes (per hr): Not documented
- Photoperiod (hrs dark / hrs light): 12 hours light/dark

IN-LIFE DATES: From: To: Not documented

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test susbtance, benzyl acetate, was mixed with olive oil to achieve concentration levels of 1000, 500, 100, 10 and 0 mg/ml

VEHICLE
- Justification for use and choice of vehicle (if other than water): olive oil
- Concentration in vehicle: Not documented
- Amount of vehicle (if gavage): Not documented
- Lot/batch no. (if required): VDR7446
- Purity: Not documented

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No information provided

Details on mating procedure:

On confirmation of the oestrous cycle of the females, 12-17 week old male rats were introduced and the males and females were co-housed from 5pm until the next morning when the presence of sperm in the vagina was considered to be successful mating. This was considered to be day zero of pregnancy. Based on their weight, pregnant rats were separated into 6 groups and relocated to separate cages.

Duration of treatment / exposure:

10 days

Frequency of treatment:

Once each day from days 6 to 15 of pregnancy

Duration of test:

10 days

No. of animals per sex per dose:

22 female rats in 1000mg/kg/day treatment group, 21 female rats in the 100 and 10 mg/kg/day treatment group and 20 female rats in the 500, 0 and non-administered treatment groups.

Control animals:

yes, concurrent no treatment

Details on study design:

No information provided.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Every 2 days

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
Examined every 2 days.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: Not documented

OTHER: The implantation in the womb, corpus lutea quantity, the implantation quantity, the resorption embryo count and the living or dead foetuses. The weight of the placenta was measured.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: No data
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: No data
- Number of late resorptions: No data
- Other:

Fetal examinations:

- External examinations: Yes: all living foetuses
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes:
- Head examinations: No data

Statistics:

One way layout dispersion method if equal dispersion detected. The Kruskal-Wallis method was used to verify significance in the case of equal dispersion. The multi-comparison verification method of Scheffe and Dunnett was used to verify the significance of subjected groups. An X2 verification method was also performed to determine the frequency of of the bone changes, internal organs of the foetuses and the gender comparison of the surviving foetuses.

Indices:

Not documented

Historical control data:

No information provided

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
There were no specific changes observed in the general condition or behaviour of the animals throughout the course of the study. No mortalities were observed in any treatment group. There were no significant differences in body weight observed between treatment groups. However, there was a tendency for the body weight to decrease in 1000mg/kg dose group from day 16 to day 20 of test substance administration and during the course of the pregnancy to day 20. There were no significant changes in food consumption in any of the treatment groups from the start to the end of pregnancy.
On gross examination, there were no visible abnormalities in the internal organs examined. There was no significant difference between the injected groups and the comparative group when the quantity of corpus lutea, the quantity of implantation, the implantation ratio and the weight of the placenta were examined.

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No significant differences were observed between the the test group and the control group when the number of surviving foetuses, the surviving ratio of implantation eggs, the number of dead foetuses, the quantitiy of resorption embryos, the resorption ration and the sex ratios were examined.
There were no significant differences between treatment and control groups when the surviving foetal weight of test animals treated with 1000mg/kg test substance was examined. However, there was a significant increase in foetal weight of the male rats injected with 100 and 10mg/kg when compared to the control group.
There were no abnormalities in the external shape of the internal organs. Onm internal inspection, all the organs examined appeared normal acorss the treatment groups when compared to the control. At the higher treatment groups of 500 and 1000mg/kg, there was a high significance of irregular shapes of organs compared to the control group. Irregularities included a light degree of ventricle expansion, levo-umbilical artery expansion and light degree of dilation of the renal pelvis. Renal pelvis dilation occurred with high frequency in significance in the 1000mg/kg treated groupcompared to the control group for the dilation of the renal pelvis.
On skeletal examination, 1 foetus displayed irregular bone shapes with coherent ribs. Bone abnormalities were observed in all test groups when compared to the controls. Bone abnormalities included wavy ribs, dumbbell shape of the thoracic vertebra body and the absence of thoracic vertebra body and the splitting of thoracic vertebra body in the chest and lumbar ribs, asence of lumber vertebra body and dumbbell shape of lumber vertebra in the lumber part. The frequency of these abnormalities was significantly high in the group treated with 1000mg/kg test substance. In the group treated with 1000mg/kg there was a significant decrease in cervical vertebra, the caudal vertebra body and the sternebrae.

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Abnormalities:

no effects observed

Developmental effects observed:

no

No additional information

Conclusions:

Based on the results of the study, the test substance, benzyl acetate, was not considered to be teratogenic under the conditions of this study. While there are indications that administration of 1000mg/kg can influence the growth of the foetus, the effects were not significant. Based on these results, the test substance dose not require classification according to Regulation EC No. 1272/2008 or Directive 67/548/EEC.

Executive summary:

In a study conducted by Ishiguro et al (1993), the test substance, benzyl acetate, was examined for its ability to induce teratogenic abnormalities when administered to female Wistar rats. The test substance was administered at dose concentrations of 0, 10, 100, 500 and 1000mg/kg to groups of 20 -22 female rats on days 6 to 15 of gestation.

There were no mortalities observed in the maternal rats and there was no effect on food consumption following test substance administration. There was a slight decrease in the body weight gain in the top dose group but it was not considered to be significant.

In the foetus, there was a significant decrease observed in the weight of the males and females exposed to the top dose level compared to the control. There was a significant increase in the occurrence of skeletal abnormalities in the foetuses exposed to the top dose level, however, as similar abnormalities were also observed at other dose levels, it was not possible to determine a concentration level.

Based on the results of the study, the test substance, benzyl acetate, was not considered to be teratogenic under the conditions of this study. While there are indications that administration of 1000mg/kg can influence the growth of the foetus, the effects were not significant. Based on these results, the test substance does not require classification according to Regulation EC No. 1272/2008 or Directive 67/548/EEC.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

250 mg/kg bw/day

Species:

rabbit

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

In a study conducted by Ishiguro et al (1993), the test substance, benzyl acetate, was examined for its ability to induce teratogenic abnormalities when administered to female Wistar rats. The test substance was administered via oral gavage at dose concentrations of 0, 10, 100, 500 and 1000 mg/kg to groups of 20 -22 female rats on days 6 to 15 of gestation.

There were no mortalities observed in the maternal rats and there was no effect on food consumption following test substance administration. There was a slight decrease in the body weight gain in the top dose group but it was not considered to be significant.

In the foetus, there was a significant decrease observed in the weight of the males and females exposed to the top dose level compared to the control. There was a significant increase in the occurrence of skeletal abnormalities in the foetuses exposed to the top dose level, however, as similar abnormalities were also observed at other dose levels, it was not possible to determine a concentration level.

Based on the results of the study, the test substance, benzyl acetate, was not considered to be teratogenic under the conditions of this study. While there are indications that administration of 1000 mg/kg can influence the growth of the foetus, the effects were not significant. Based on these results, the test substance dose not require classification according to Regulation EC No. 1272/2008 or Directive 67/548/EEC.

In a separate study published by the FDA (1972), the developmental toxicity of sodium benzoate was assessed in three rodent species (rat, mouse and hamster) and a non-rodent species (rabbit). The data were consistent among the species. The data from the rabbit study are presented. The study showed no maternal or developmental toxicity at dose levels of up to 250 mg/kg bw/day.

Justification for classification or non-classification

There is no indication that benzyl acetate is a reproductive toxin based on read-across studies. Classification according to the CLP Regulation is not required.