Isopropyl benzoate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

toxicity to reproduction

Remarks:

other: 4 generation study

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1960-2001

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

This is a summary of studies reviewed by the expert groups of the OECD High Production Volume (HPV) Chemicals Programme (SIDS Review Committee), JECFA and the U.S. Cosmetic Ingredient Review Expert Panel (CIR), which accepted the data as scientifically valid.. Documentation details may be missing as the studies were carried out some years ago and did not always fulfill present-day guidelines.

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD guideline 416

GLP compliance:

not specified

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: feed

Details on exposure:

0.5 or 1 % in diet (approx. 375 or 750 mg/kg/day)

Analytical verification of doses or concentrations:

not specified

Remarks:

Doses / Concentrations:
375 and 750 mg/kg/day
Basis:
nominal in diet
20 rats were fed contineuously with 375 or 750 mg/kg bw

No. of animals per sex per dose:

20 Rats per sex/ group

Control animals:

yes, plain diet

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: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

In all 4 generations, no effects on fertility (“Fortpflanzung”) and lactation (“Aufzugt der Jungen”) were found.

Key result

Dose descriptor:

NOAEL

Remarks:

750

Effect level:

>= 750 - <= 800 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

Key result

Dose descriptor:

NOAEL

Effect level:

>= 750 - ca. 800 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

No effect was found in all 4 generations under this study.

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

>= 750 - 800 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

Reproductive effects observed:

not specified

Key result

Reproductive effects observed:

no

In all 4 generations no influence on growth (weight, weight gain and food efficiency and organ weights was found. In all 4 generations no effects on fertility (Forzplanzung) and lactation (Aufzugtder Jungen) was found.In addition aso-called "Alters Paarung" after 48 weeks gave no influence on start of menopause.

Conclusions:

Benzoic acid was administered in the diet to rats for 4 consecutive generations. In all 4 generations, no effects on fertility and lactation were found. The substance is not a reproductive toxicant. This study is informative for evaluation of the toxicity of members of the Alkyl Benzoates category, and is adequate for filling the data requirement for the registration of this substance. It is valid for hazard classification and risk assessment.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

adequate

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

(discussed below).

Short description of key information:
No reproductive toxicity was observed in a 4-generation chronic study on benzoic acid.

Justification for selection of Effect on fertility via oral route:
experimental data

Effects on developmental toxicity

Description of key information

No developmental toxicity was observed in a 4-generation chronic study on benzoic acid.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1960-2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data reviewed by an expert panel of the OECD HPV Programme and considered "valid with restrictions." "Meets generally accepted scientific standards, well documented and acceptable for assessment. Flagged as a critical study for SIDS endpoint."

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD 416 Two-Generation Reproduction Toxicity

Deviations:

yes

Remarks:

Two doses (plus control) only, extends to 4 generations

GLP compliance:

not specified

Limit test:

yes

Species:

rat

Route of administration:

oral: feed

Details on exposure:

First 8 weeks: paired feed technique; afterwards, ad libitum.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No details provided. The mean compound consumption was calculated according to Lehman AJ, Food Drug Official Q. Bull., 66 (1954).

Duration of treatment / exposure:

Generations 1 & 2: lifelong.
Generation 3: 16 weeks
Generation 4: until breeding.

Frequency of treatment:

continuously in diet

Duration of test:

4 generations (48 weeks)

No. of animals per sex per dose:

20 per sex per dose group. Initial body weight: 40-50 g.

Control animals:

yes

Details on study design:

Group I was dosed with 375 mg/kg.
Group II was dosed with 750 mg/kg;
An additional control group was used.

Maternal examinations:

DETAILED CLINICAL OBSERVATIONS: Yes
BODY WEIGHT: Yes
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation after 21 days 3rd generation
- Organs examined: Live fetuses were removed, examined for gross malformations, weighed, and prepared for histological examination. Skeletal examination was carried out under low magnification.

Fetal examinations:

- External examinations: Yes:
- Head examinations: No data

Statistics:

No abnormalities survivors rates as comparable to the control animals.

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
In all 4 generations, no influence on growth (weight, weight gain and food efficiency (measured by protein efficiency)) and organ weights was found. In all 4 generations, no effects on fertility and lactation were found. No remarkable histopathological findings were found in the 3rd generation animals. In the paper no information is given on the organs investigated, however the robustness of the total study, the reputation of the investigators, as well as the reputation of the Professor who did the histopathologic investigation, a high quality has to be assumed. In addition after 48 weeks, there was no influence on start of menopause. Feeding of 375 mg/kg bw/d led to prolongation of survival compared to controls.
NOAEL (Parental) > 750 mg/kg/day

Dose descriptor:

NOAEL

Effect level:

>= 500 - 750 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

500 - 750 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
NOAEL (F1 Offspring) > 750 mg/kg/day
NOAEL (F2 Offspring) > 750 mg/kg/day

Dose descriptor:

NOAEL

Effect level:

> 750 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No fetal effects observed.

Remarks on result:

not determinable due to absence of adverse toxic effects

Abnormalities:

not specified

Developmental effects observed:

not specified

There are differing estimates of the dose as expressed in mg/kg bw/d, which range from 500 to 750. See discussion in Kieckebush and Lang, 1960, endpoint study record under Repeated Dose Toxicity.

Conclusions:

In a 4-generation study (48 weeks) reproductive toxicity study of 0.5 and 1% benzoic acid in rats by the dietary route, there were no observed adverse effects in growth, body weight, organ weights, reproductive parameters or developmental effects. The Parental and offspring NOAELs were greater than 500-750 mg/kg bw/d. This study is informative for evaluation of the toxicity of members of the Alkyl Benzoates category, and is adequate for filling the data requirement for the registration of this substance. It is valid for hazard classification and risk assessment.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

adquate

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

There are several reproductive toxicity studies of benzoic acid (Kieckebush and Lang, 1960, Kimmel et al., 1971) and sodium benzoate (Mortgareidge (U.S. FDA), 1972) which indicate that oral administration of these substances does not result in reproductive toxicity to rats. The studies have been evaluated as valid and supportive of benzoic acid and salts as food additives (JECFA, beginning in 1961; U.S. FDA). There was no maternal toxicity nor adverse reproductive outcomes among rats in 4 generations of dietary exposure to 1% benzoic acid (Kieckebush and Lang, 1960), equivalent to approximately 500-750 mg/kg bw/d. The maternal and offspring NOAELs are established 500 mg/kg bw/d, supported by the findings of Kimmel, et al, 1971 at 510 mg/kg bw/d, and Mortgareidge, 1972, at 175 mg/kg bw/d. The Scientific Committee on Food (SCF) in 2002 reviewed the available data and concluded that the data were adequate and that no additional reproductive/developmental toxicity studies on benzoic acid were indicated. Many expert toxicology groups have made the same conclusion.

Isopropyl benzoate is part of a category of chemicals, the Alkyl Benzoates, which utilize this data on benzoic acid and sodium benzoate for safety evaluation. The establishment of a benzoates category has been accepted by numerous expert toxicology groups, including the Joint Expert Committee for Food Additives (JECFA), the Scientific Committee on Food (SCF), the European Food Safety Organization (EFSA), the U.S. Food and Drug Administration (FDA), the World Health Organization (WHO), and the OECD High Production Volume Chemicals Programme (OECD HPV). The basis of the large category for all these organizations (EFSA now has a category of 41 substances qualifying as Benzyl Derivative flavours) is a common breakdown product, benzoic acid, which forms rapidly by acidic hydrolysis as well as due to the action of esterase enzymes. Genetic and reproductive toxicity studies are also negative. In addition to this category, the U.S. Cosmetic Ingredient Review (CIR) Expert Panel evaluated the safety of 17 alkyl benzoate esters as a category (2012) based on a common functional group (benzoic acid ester) which is metabolized to benzoic acid.

For the alcohol metabolite, isopropanol, reproductive and developmental toxicity was investigated in rats in several guideline studies (one- and two-generation reproductive toxicity protocols) and also in a developmental neurotoxicity study (Bates et al., 1994). There were no significant findings specific to reproduction, and no evidence of adverse effects in offspring at doses which were not toxic to the dams. The alcohol, likely via central nervous system depression, resulted in decreased mating indices (Beyer et al., 1993). High doses resulted in death, increased liver and kidney weights and liver pathology. The parental NOAEL in the two-generation study was 500 mg/kg bw/d, and for offspring, 1000 mg/kg bw/d. The parental NOAEL in the developmental neurotoxicity was 700 mg/kg bw/d, and for offspring, 1000 mg/kg bw/d.

The NOAEL of 500 mg/kg bw/d is adequate to address the endpoints of reproductive and developmental toxicity.

Justification for selection of Effect on developmental toxicity: via oral route:
experimental data

Justification for classification or non-classification

As no reproductive and developmental toxicity was observed with metabolites of isopropyl benzoate, the criteria for classification and labelling according to Regulation EC No. 1272/2008 are not met.