Registration Dossier

Administrative data

Endpoint:
multi-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Publication includes a well documented study summary. For justification of read across see section 13.

Data source

Reference
Reference Type:
publication
Title:
Die Verträglichkeit der Benzoesäure im chronischen Fütterungsversuch
Author:
Kieckebusch W., Lang K.
Year:
1960
Bibliographic source:
Arzneimittel Forschung 10, 1001-1003

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
4-generation study
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Farbwerke Bayer (Elberfeld)
- Weight at study initiation: 40-50 g
- Diet: during the first 8 weeks the rats were fed according to the pair-feeding technique, afterwards food was available ad libitum
- Water: ad libitum

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): no data
Details on mating procedure:
- M/F ratio per cage: 1 / 1
- Length of cohabitation: 14 d
- Proof of pregnancy: no data
- Further matings after two unsuccessful attempts: yes
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The doses were analytically verified at intervalls of six weeks.
Duration of treatment / exposure:
The first two generations were treated lifelong, the 3rd generation for 16 weeks and the 4th generation until breeding.
Frequency of treatment:
During the first 8 weeks the rats were fed according to the pair-feeding technique, afterwards food was available ad libitum.
Details on study schedule:
no data
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 0.5, 1 % (0, 250, 500 mg/kg bw)
Basis:
nominal in diet
No. of animals per sex per dose:
20
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: no data
Positive control:
no data

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: weekly during the first 8 weeks and at four weeks intervalls afterwards

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE: No data
Oestrous cyclicity (parental animals):
no data
Sperm parameters (parental animals):
no data
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring: total number of offspring, % of surviving offspring

Postmortem examinations (parental animals):
no data
Postmortem examinations (offspring):
no data
Statistics:
no data
Reproductive indices:
no data
Offspring viability indices:
no data

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed

Details on results (P0)

ORGAN WEIGHTS (PARENTAL ANIMALS)
An increased liver weigth was observed in females of the 3rd generation but, this increase was not treatment-related. Liver weigt increases were supposed to be caused by a weight increase during lactation and variation in litter sizes.

Effect levels (P0)

Dose descriptor:
NOEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Intake of 150 mg (1%) of benzoic acid per day per rat and a bodyweight of 300 g per rat were assumed.

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Effect levels (F1)

Dose descriptor:
NOEL
Generation:
F1
Effect level:
500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Rats were exposed to 0.5 and 1 % benzoic acid in a chronic feeding study. 1 % benzoic acid was equivalent to 500 mg/kg bw/day. As no effects were observed in any of the four generations, the NOEL was determined to be 500 mg/kg bw.
Executive summary:

Rats were exposed to 0.5 and 1 % benzoic acid in two groups of 20 male and 20 female rats in a chronic feeding study. 1 % benzoic acid in the food is equivalent to 500 mg/kg bw/day. The negative control group 1 consisted of the same amount of animals and received untreated food. The first two generations were treated lifelong, the 3rd generation was treated for 16 weeks and the 4th generation until breeding. No effects were observed in any of the generations except for a liver weight increase in the females of the 3rd generation, but this increase was not considered to be caused by the treatment. On the basis of these results the NOEL for benzoic acid was concluded to be 500 mg/kg bw.