Aluminium tribenzoate

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

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

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Reference

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

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

A feeding study was performed, in which 4 generations of rats received benzoic acid (0, 0.5 or 1%). Body weight development was determined for each generation. Reproduction parameters were assessed (number of pups, survival of pups, fertility, litter size). Rats of the F2 generation were sacrificed at 16 weeks of age. Weights of brain, heart, liver, spleen, kidneys and testes were measured. Histopathological examination was performed (F2 generation).

GLP compliance:

no

Remarks:

test performed before GLP principles were implemented

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bayer
- Females nulliparous and non-pregnant: yes
- Age at study initiation: not specified
- Weight at study initiation: 40-50 g
- Housing: double cages
- Diet: Standard rodent diet (Latz, Euskirchen), first 8 weeks "paired feed"-technique, otherwise ad libitum
- Water: ad libitum

Route of administration:

oral: feed

Vehicle:

not specified

Details on mating procedure:

At the age of 11-12 weeks, one male and one female from the same dose group were paired. The male and female were co-housed for 2 weeks. In case the female was not pregnant, this pocedure was repeated after 8 weeks. The females were paired at 48 weeks to detect delay in menopause. After successful mating, pups were counted on day 1, 2 and 21.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

4 generations

Frequency of treatment:

Continuous exposure through feed

Dose / conc.:

0.5 other: %

Remarks:

Corresponds to appr. 200 mg/kg bw/day

Dose / conc.:

1 other: %

Remarks:

Corresponds to appr. 400 mg/kg bw/day

No. of animals per sex per dose:

20

Control animals:

yes

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Not specified

BODY WEIGHT: Yes
- Time schedule for examinations: weekly until 8 weeks of age, otherwise monthly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

Postmortem examinations (parental animals):

The F2 rats were sacrificed after week 16 and examined histopathologically. Organ weights were measured (brain, heart, liver, spleen, kidneys and testes).

Reproductive indices:

Number of pregnant females was recorded (all generations).

Offspring viability indices:

The number of live pups on PND 1, 2 and 21 were recorded (all generations).

Mortality:

no mortality observed

Description (incidence):

The average survival (F0 and F1-generation) was slightly increased in the dosed groups compared to the control groups (average lifespan in days: 785 ± 20, 899 ± 28 and 827 ± 25 for the control group and the groups dosed at 0.5% and 1,0%, respectively).

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Reproductive performance:

no effects observed

Description (incidence and severity):

The percentage of sterile females was 15%, 10% and 4% for the control group and the groups exposed to 0.5% and 1% in feed, respectively. The average number of pups per nest was 9.0 ± 0.33, 9.5 ± 0.26 and 9.6 ± 0.29 (after first mating, all generations) and 6.9, 7.7 and 7.5 (second mating, F1 only) for the control group and the groups exposed to 0.5% and 1% in feed, respectively. The survival of pups after the first three weeks was slightly lower for the exposed groups (74%, 66% and 65% (after first mating, all generations) and 72%, 61% and 73% (second mating, F1 only) for the control group and the groups exposed to 0.5% and 1% in feed, respectively). This is expected to be related to the higher number of pups, and not to the exposure to the test item. There was no effect seen on onset of menopause.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects seen up to and including 1% benzoic acid in feed.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 400 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects seen up to and including the highest dose tested.

Key result

Critical effects observed:

no

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

The survival of pups after the first three weeks was slightly lower for the exposed groups (74%, 66% and 65% (after first mating, all generations) and 72%, 61% and 73% (second mating, F1 only) for the control group and the groups exposed to 0.5% and 1% in feed, respectively). This is considered to be related to the higher number of pups, and not to the exposure to the test item.

Key result

Dose descriptor:

NOAEL

Generation:

other: F0, F1, F2 and F3

Effect level:

>= 1 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects seen up to and including the highest dose tested.

Key result

Dose descriptor:

NOAEL

Generation:

other: F0, F1, F2 and F3

Effect level:

>= 400 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects seen up to and including the highest dose tested.

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Conclusions:

Based on the results of a feeding study with benzoic acid over 4 generations, the NOAEL for reproduction toxicity was established to be at least 1% in feed, which correlates to approximately 400 mg/kg bw/day.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1960

Report date:

1960

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

A feeding study was performed, in which 4 generations of rats received benzoic acid (0, 0.5 or 1%). Body weight development was determined for each generation. Reproduction parameters were assessed (number of pups, survival of pups, fertility, litter size). Rats of the F2 generation were sacrificed at 16 weeks of age. Weights of brain, heart, liver, spleen, kidneys and testes were measured. Histopathological examination was performed on rats of F2 generation.

GLP compliance:

no

Remarks:

test performed before GLP principles were implemented

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Benzoic acid

EC Number:

200-618-2

EC Name:

Benzoic acid

Cas Number:

65-85-0

Molecular formula:

C7H6O2

IUPAC Name:

benzoic acid

Test animals

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bayer
- Females nulliparous and non-pregnant: yes
- Age at study initiation: not specified
- Weight at study initiation: 40-50 g
- Housing: double cages
- Diet: Standard rodent diet (Latz, Euskirchen), first 8 weeks "paired feed"-technique, otherwise ad libitum
- Water: ad libitum

Administration / exposure

Route of administration:

oral: feed

Vehicle:

not specified

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

4 generations

Frequency of treatment:

Continuous exposure through feed

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0.5 other: %

Doses / concentrations 2

Dose / conc.:

1 other: %

No. of animals per sex per dose:

20

Details on study design:

At the age of 11-12 weeks, one male and one female from the same dose group were paired. The male and female were co-housed for 2 weeks. In case the female was not pregnant, this pocedure was repeated after 8 weeks. The females were paired at 48 weeks to detect delay in menopause. After successful mating, pups were counted on day 1, 2 and 21.
The F0 and F1 rats were observed until death. The F2 rats were sacrificed after week 16 and examined histopathologically. Organ weights were measured (brain, heart, liver, spleen, kidneys and testes). The F3 rats were sacrificed after week 16.

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Not specified

BODY WEIGHT: Yes
- Time schedule for examinations: weekly until 8 weeks of age, otherwise monthly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

Results and discussion

Results of examinations

Mortality:

no mortality observed

Description (incidence):

The average survival (F0 and F1-generation) was slightly increased in the dosed groups compared to the control groups (average lifespan in days: 785 ± 20, 899 ± 28 and 827 ± 25 for the control group and the groups dosed at 0.5% and 1,0%, respectively).

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects seen up to and including 1% benzoic acid in feed.

2

Key result

Dose descriptor:

NOAEL

Effect level:

>= 400 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects seen up to and including highest dose tested.

Target system / organ toxicity

Critical effects observed:

no

Any other information on results incl. tables

Based on the default values as proposed in TNO V98 report the average body weight rat in a chronic study is appr. 375 g (male and female). According to the authors, the average intake was approximately 150 mg benzoic acid/ rat, which corresponds to an exposure of appr. 400 mg/kg bw/day.

Applicant's summary and conclusion

Conclusions:

Based on the results of a feeding study with benzoic acid over 4 generations, the NOAEL was established to be at least 1% in feed, which correlates to approximately 400 mg/kg bw/day.