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Toxicological information

Toxicity to reproduction

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Endpoint:
reproductive toxicity, other
Remarks:
Evaluation of Rodent Sperm, Vaginal Cytology, and Reproductive Organ Weight Data from National Toxicology Program 13-Week Studies
Type of information:
experimental study
Remarks:
In a chronic feeding study, male and female rats were exposed to up to 1% benzoic acid in the diet through four generations.
Adequacy of study:
key study
Study period:
Publication compiles reproductive parameters from 13-week repeated dose studies from 50 different substances.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data are derived from National Toxicology Program (NTP) testing program.
Justification for type of information:
See Read Across Justification document in Section 13.2 of IUCLID
Cross-reference
Reason / purpose:
read-across: supporting information
Reference
Endpoint:
reproductive toxicity, other
Remarks:
Evaluation of Rodent Sperm, Vaginal Cytology, and Reproductive Organ Weight Data from National Toxicology Program 13-Week Studies
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Remarks:
In a chronic feeding study, male and female rats were exposed to up to 1% benzoic acid in the diet through four generations.
Adequacy of study:
key study
Study period:
Publication compiles reproductive parameters from 13-week repeated dose studies from 50 different substances.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data are derived from National Toxicology Program (NTP) testing program.
Justification for type of information:
See Read Across Justification document in Section 13.2 of IUCLID
Reason / purpose:
read-across source
Qualifier:
no guideline followed
Principles of method if other than guideline:
The studies were designed as standard repeated-dose (13-week) and carcinogenicity (2-year) assays, commensurate with accepted standards at the time.

Data are derived from National Toxicology Program series of 13-week repeated dose studies. The publication states that "Sperm morphology and vaginal cytology examinations (SMVCEs), which include evaluations of motility, concentration and head morphology of sperm from the cauda epididymis, and male reproductive organ weight data, were developed by the National Toxicology Program as a screening system for reproductive toxicants.
GLP compliance:
yes
Remarks:
GLP Compliance confirmed in NTP TR 431
Limit test:
no
Justification for study design:
The studies were designed as standard repeated-dose (13-week) and carcinogenicity (2-year) assays, commensurate with accepted standards at the time.
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Benzyl acetate was obtained in two lots (8743-84 and 845585) from GivaudanCorporation (Clifton, NJ). Identity and purity analyses were conducted by the analytical chemistry laboratory, Midwest Research Institute (Kansas City, MO), and confirmed by the study laboratory. Purity was determined to be > 98% for both lots.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL: Stability studics performed at the analytical chemistry laboratory indicated that benzyl acetate was stable as a bulk chemical for 2 weeks at temperatures as high as 60" C. The stability of the bulk chemical was monitorcd periodically by the study laboratory using infrarcd and spectroscopy gas ultraviolet and chromatography; no change in purity was observed.
Species:
other: both rat and mouse are reported by Morrissey et al.
Strain:
other: F344/N rats; B6C3F1 mice
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Frederick Cancer Research Facility (Frederick, MD)
- Age at study initiation: Average of 30 days old.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days prior to study initiation
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.6 +/- 3.0 degrees C
- Humidity (%): 46-65%
- Air changes (per hr): Minimum of 10 changes/hour.
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark
Route of administration:
oral: feed
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): diet with test material prepared weekly.
- Storage temperature of food: Dose formulations stored at -20 degrees C in sealed, double plastic bags.
Details on mating procedure:
Not Applicable: The study is a 13-week repeated dose study, and is included regarding the collection of data on effects on reproductive parameters, not including mating.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
"Periodic analyses of the dose formulations of benzyl acctate were conducted at the sludy laboratory and the analytical chemistry laboratory using gas chromatography. The stability of 330ppm dose formulations stored in the dark at -20 degrees C was established for at least 3 weeks. Dose formulations were analyzed four times for the 13-week studies and approximately every 6 to 8 weeks during the 2-year studies. All dose formulations for rats and mice were within 10% of the target concentrations throughout the studies." "The results of periodic referee analysis pcrformed by the analytical chemistry laboratory indicated agreement with the results obtained by the study laboratory."
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Feed containing test material provided ad libitum.
Details on study schedule:
The appropriate feed was supplied every 1 to 2 days. Feed and water were available ad libitum. Feed consumption was recorded daily by cage. Rats were housed five per cage and mice were housed individually. Clinical findings were recorded once weekly. The animals were weighed at the beginning of the studies, weekly, and at the end of the studies.
Dose / conc.:
0 ppm
Remarks:
Control for both rats and mice
Dose / conc.:
3 130 ppm
Remarks:
Resulted in 235 mg/kg bw/day in rats (average of Male and Female), and 537.5 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
6 250 ppm
Remarks:
Resulted in 470 mg/kg bw/day in rats (average of Male and Female), and 1,140 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
12 500 ppm
Remarks:
Resulted in 915 mg/kg bw/day in rats (average of Male and Female), and 2,490 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
25 000 ppm
Remarks:
Resulted in 1,810 mg/kg bw/day in rats (average of Male and Female), and 4,000 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
50 000 ppm
Remarks:
Resulted in 4,200 mg/kg bw/day in rats (average of Male and Female), and 8,650 mg/kg bw/day in mice (average of Male and Female).
No. of animals per sex per dose:
10 males/group
10 females/group
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Not described in NTP report.
Positive control:
None stated
Parental animals: Observations and examinations:
N/A - This study endpoint record is focused on reproductive parameters as reported by Morrissey et al., 1988.
Oestrous cyclicity (parental animals):
Estrous cyclicity evaluated in mice and rats.
Sperm parameters (parental animals):
Sperm quality parameters were evaluated at the end of the 13-week in-life phase, including:
- Sperm motility
- Sperm count
- Sperm head staining for morphology
Litter observations:
Not examined in this study.
Postmortem examinations (parental animals):
Regarding male reproductive tissues, in the basic NTP study, the right testis and seminal vesicle was necropsied, and histopathology was examined on testis with epididymis, and seminal vesicle. For testis w/ epididymis, all doses were evaluated, not just the two high doses. Regarding female reproductive tissues, in the basic NTP study, the uterus was necropsied, and histopathology of the uterus was examined.
Postmortem examinations (offspring):
N/A - This study endpoint record is focused on reproductive parameters as reported by Morrissey et al., 1988.
Statistics:
For body, organ and relative organ weight, either William's or Dunnett's test was used. Because Dunnett's test makes no allowance for dose-response relationships, Jonckheere's test (a non-parametric test for dose-response relationship) was used to determine whether Dunnett's or Williams' would be used. Potential decreases in sperm motility and increases in percentage abnormal sperm were evaluated using Jonckheere's test. Sperm density was evaluated using the Kruskal-Wallis test.
Reproductive indices:
Not examined in this study.
Offspring viability indices:
Not examined in this study.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Overall toxicity findings were not discussed in Morrissey et al. (1988), but were discussed in the original NTP study. The discussion of clinical findings in rats was limited to the description of "tremors, ataxia and urine stains" in the high-dose animals. The discussion of clinical findings in mice was as follows:
"The significant clinical finding was tremors, which occurred only in females and was predominant in the 50,000 ppm group; however, one 12,500
ppm female and one 25,000 ppm female also exhibited tremors. The tremors occurred first on day 16 of the study in three females receiving 50,000
ppm, day 94 in one female receiving 25,000 ppm, and day 93 in one female receiving 12,500 ppm, and continued until the end of the study."
Mortality:
mortality observed, treatment-related
Description (incidence):
In rats, nine male and nine female animals receiving the highest dose died or were killed moribund between weeks 2 and 8 of the study. One 12,500 ppm female died under anesthesia during blood collection at the end of the study.

In mice, one 50,000 ppm male mouse died, and one 50,000 ppm female mouse was sacrificed as moribund before the end of the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In rats, the mean body weight gain and the final mean body weight of 25,000 ppm males were significantly lower than control. The final mean body weight of 25,000 ppm males was 10% lower than that of controls, whereas the final body weights of the surviving 50,000 ppm male and female animals was less than 50% of controls. Final mean body weights of males and females of other exposed groups were similar to or slightly lower than those of controls.

In mice, statistically significant dose-related decreases in final mean body weights occurred in all groups of exposed male and female mice.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In rats, feed consumption was comparable to control in all but the 25,000 and 50,000 ppm groups, where reduced consumption was attributed to decreased palatability and/or toxicity.

In mice, the mean feed consumption was lower in all groups when compared to control.
Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Reproductive effects observed:
no
Lowest effective dose / conc.:
50 000 ppm
Treatment related:
no

This citation is being presented to cover apparent gaps in the Kieckebusch and Lang (1960) study regarding effects on reproducitve tissues/organs. Table 1 of the Morrissey et al. paper provides the following information regarding benzyl acetate on male reproductive tissues, relative to control:

Species

Terminal Body Weight

Organ Weight

Sperm Parameters

R. cauda

R. epididymis

R. Testis

Absolute

Relative

Absolute

Relative

Absolute

Relative

Motility (%)

Density

Abnormal (%)

Mouse

Decrease

Decrease

Increase

Decrease

Increase

Decrease

Increase

No difference

No difference

No difference

Rat

Decrease

No difference

No difference

No difference

No difference

No difference

No difference

No difference

No difference

No difference

Regarding female reproductive tissue outcomes, benzyl acetate was noted to significantly increase the mean cycle length in the high-dose group of mice.

Conclusions:
These data supplement missing endpoints from the Kieckebusch and Lang (1960) Key Study, and fill the gap in that study design. In the National Toxicology Program (Morrissey et al., 1988) 13-week feeding benzyl acetate studies in rats and mice, both sperm parameters (epididymis, cauda epididymis, testis weights, and sperm motility, density and percent abnormal sperm) and estrus cyclicity measurements were collected. Dose levels of 2000 to 3000 mg/kg/day did not affect any endpoints of male reproduction. Estrus cycles were lengthened in female mice receiving >3000 mg/kg/day but this only occurred at a dose levels also causing significant decreases in growth (body weights and body weight gain).
Executive summary:

Regulation (EC) No 1907/2006 (REACH), Annex XI 1.5., establishes, that substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group of substances. The similarity may also be based on the likelihood of common breakdown products via physical and biological processes. Given that the grouping of substances and read-across approach can be justified, data gaps may be fulfilled from data of reference substance(s) within the group. This avoids the need to test every substance for every endpoint, which is highly recommended in the context of animal welfare considerations (Directive 2010/63/EU, 2010).

 

For sodium benzoate, the fertility and developmental toxicity endpoints are filled by read across from a group of structurally similar substances.

 

Benzyl acetate, benzyl alcohol, benzaldehyde, and benzoic acid and its sodium and potassium salt were considered as a single category regarding human health by JECFA as they are all rapidly metabolized and excreted via a common pathway within 24 hrs (JECFA 1997).

 

The data presented below if for benzyl acetate using the read across justification described above and is used as read across for sodium benzoate as requested by ECHA in its letter ‘’Submission FP609849-12 – request for dossier update’’ received by the lead registrant on October 03, 2019 and attached in Section 13 of the IUCLID dossier. 

These data supplement missing endpoints from the Kieckebusch and Lang (1960) Key Study, and fill the gap in that study design. In the National Toxicology Program (Morrissey et al., 1988) 13-week feeding benzyl acetate studies in rats and mice, both sperm parameters (epididymis, cauda epididymis, testis weights, and sperm motility, density and percent abnormal sperm) and estrus cyclicity measurements were collected. Dose levels of 2000 to 3000 mg/kg/day did not affect any endpoints of male reproduction. Estrus cycles were lengthened in female mice receiving >3000 mg/kg/day but this only occurred at a dose levels also causing significant decreases in growth (body weights and body weight gain).

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Evaluation of rodent sperm, vaginal cytology, and reproductive organ weight data from National Toxicology Program 13-week studies
Author:
Morrissey, R.E., Schwetz, B., Lamb IV, J.C., Ross, M.D., Teague, J.L. and Morris, R.W.
Year:
1988
Bibliographic source:
Fundam. Appl. Toxicol., 11, 343-358
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
The studies were designed as standard repeated-dose (13-week) and carcinogenicity (2-year) assays, commensurate with accepted standards at the time.

Data are derived from National Toxicology Program series of 13-week repeated dose studies. The publication states that "Sperm morphology and vaginal cytology examinations (SMVCEs), which include evaluations of motility, concentration and head morphology of sperm from the cauda epididymis, and male reproductive organ weight data, were developed by the National Toxicology Program as a screening system for reproductive toxicants.
GLP compliance:
yes
Remarks:
GLP Compliance confirmed in NTP TR 431
Limit test:
no
Justification for study design:
The studies were designed as standard repeated-dose (13-week) and carcinogenicity (2-year) assays, commensurate with accepted standards at the time.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
Benzyl acetate was obtained in two lots (8743-84 and 845585) from GivaudanCorporation (Clifton, NJ). Identity and purity analyses were conducted by the analytical chemistry laboratory, Midwest Research Institute (Kansas City, MO), and confirmed by the study laboratory.
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Benzyl acetate was obtained in two lots (8743-84 and 845585) from GivaudanCorporation (Clifton, NJ). Identity and purity analyses were conducted by the analytical chemistry laboratory, Midwest Research Institute (Kansas City, MO), and confirmed by the study laboratory. Purity was determined to be > 98% for both lots.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL: Stability studics performed at the analytical chemistry laboratory indicated that benzyl acetate was stable as a bulk chemical for 2 weeks at temperatures as high as 60" C. The stability of the bulk chemical was monitorcd periodically by the study laboratory using infrarcd and spectroscopy gas ultraviolet and chromatography; no change in purity was observed.

Test animals

Species:
other: both rat and mouse are reported by Morrissey et al.
Strain:
other: F344/N rats; B6C3F1 mice
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Frederick Cancer Research Facility (Frederick, MD)
- Age at study initiation: Average of 30 days old.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days prior to study initiation
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.6 +/- 3.0 degrees C
- Humidity (%): 46-65%
- Air changes (per hr): Minimum of 10 changes/hour.
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark

Administration / exposure

Route of administration:
oral: feed
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): diet with test material prepared weekly.
- Storage temperature of food: Dose formulations stored at -20 degrees C in sealed, double plastic bags.
Details on mating procedure:
Not Applicable: The study is a 13-week repeated dose study, and is included regarding the collection of data on effects on reproductive parameters, not including mating.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
"Periodic analyses of the dose formulations of benzyl acctate were conducted at the sludy laboratory and the analytical chemistry laboratory using gas chromatography. The stability of 330ppm dose formulations stored in the dark at -20 degrees C was established for at least 3 weeks. Dose formulations were analyzed four times for the 13-week studies and approximately every 6 to 8 weeks during the 2-year studies. All dose formulations for rats and mice were within 10% of the target concentrations throughout the studies." "The results of periodic referee analysis pcrformed by the analytical chemistry laboratory indicated agreement with the results obtained by the study laboratory."
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Feed containing test material provided ad libitum.
Details on study schedule:
The appropriate feed was supplied every 1 to 2 days. Feed and water were available ad libitum. Feed consumption was recorded daily by cage. Rats were housed five per cage and mice were housed individually. Clinical findings were recorded once weekly. The animals were weighed at the beginning of the studies, weekly, and at the end of the studies.
Doses / concentrationsopen allclose all
Dose / conc.:
0 ppm
Remarks:
Control for both rats and mice
Dose / conc.:
3 130 ppm
Remarks:
Resulted in 235 mg/kg bw/day in rats (average of Male and Female), and 537.5 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
6 250 ppm
Remarks:
Resulted in 470 mg/kg bw/day in rats (average of Male and Female), and 1,140 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
12 500 ppm
Remarks:
Resulted in 915 mg/kg bw/day in rats (average of Male and Female), and 2,490 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
25 000 ppm
Remarks:
Resulted in 1,810 mg/kg bw/day in rats (average of Male and Female), and 4,000 mg/kg bw/day in mice (average of Male and Female).
Dose / conc.:
50 000 ppm
Remarks:
Resulted in 4,200 mg/kg bw/day in rats (average of Male and Female), and 8,650 mg/kg bw/day in mice (average of Male and Female).
No. of animals per sex per dose:
10 males/group
10 females/group
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Not described in NTP report.
Positive control:
None stated

Examinations

Parental animals: Observations and examinations:
N/A - This study endpoint record is focused on reproductive parameters as reported by Morrissey et al., 1988.
Oestrous cyclicity (parental animals):
Estrous cyclicity evaluated in mice and rats.
Sperm parameters (parental animals):
Sperm quality parameters were evaluated at the end of the 13-week in-life phase, including:
- Sperm motility
- Sperm count
- Sperm head staining for morphology
Litter observations:
Not examined in this study.
Postmortem examinations (parental animals):
Regarding male reproductive tissues, in the basic NTP study, the right testis and seminal vesicle was necropsied, and histopathology was examined on testis with epididymis, and seminal vesicle. For testis w/ epididymis, all doses were evaluated, not just the two high doses. Regarding female reproductive tissues, in the basic NTP study, the uterus was necropsied, and histopathology of the uterus was examined.
Postmortem examinations (offspring):
N/A - This study endpoint record is focused on reproductive parameters as reported by Morrissey et al., 1988.
Statistics:
For body, organ and relative organ weight, either William's or Dunnett's test was used. Because Dunnett's test makes no allowance for dose-response relationships, Jonckheere's test (a non-parametric test for dose-response relationship) was used to determine whether Dunnett's or Williams' would be used. Potential decreases in sperm motility and increases in percentage abnormal sperm were evaluated using Jonckheere's test. Sperm density was evaluated using the Kruskal-Wallis test.
Reproductive indices:
Not examined in this study.
Offspring viability indices:
Not examined in this study.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Overall toxicity findings were not discussed in Morrissey et al. (1988), but were discussed in the original NTP study. The discussion of clinical findings in rats was limited to the description of "tremors, ataxia and urine stains" in the high-dose animals. The discussion of clinical findings in mice was as follows:
"The significant clinical finding was tremors, which occurred only in females and was predominant in the 50,000 ppm group; however, one 12,500
ppm female and one 25,000 ppm female also exhibited tremors. The tremors occurred first on day 16 of the study in three females receiving 50,000
ppm, day 94 in one female receiving 25,000 ppm, and day 93 in one female receiving 12,500 ppm, and continued until the end of the study."
Mortality:
mortality observed, treatment-related
Description (incidence):
In rats, nine male and nine female animals receiving the highest dose died or were killed moribund between weeks 2 and 8 of the study. One 12,500 ppm female died under anesthesia during blood collection at the end of the study.

In mice, one 50,000 ppm male mouse died, and one 50,000 ppm female mouse was sacrificed as moribund before the end of the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In rats, the mean body weight gain and the final mean body weight of 25,000 ppm males were significantly lower than control. The final mean body weight of 25,000 ppm males was 10% lower than that of controls, whereas the final body weights of the surviving 50,000 ppm male and female animals was less than 50% of controls. Final mean body weights of males and females of other exposed groups were similar to or slightly lower than those of controls.

In mice, statistically significant dose-related decreases in final mean body weights occurred in all groups of exposed male and female mice.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In rats, feed consumption was comparable to control in all but the 25,000 and 50,000 ppm groups, where reduced consumption was attributed to decreased palatability and/or toxicity.

In mice, the mean feed consumption was lower in all groups when compared to control.

Effect levels (P0)

Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects

Results: P1 (second parental generation)

Effect levels (P1)

Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects

Results: F1 generation

Effect levels (F1)

Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects

Results: F2 generation

Effect levels (F2)

Key result
Dose descriptor:
NOEL
Effect level:
> 50 000 ppm
Remarks on result:
not determinable due to absence of adverse toxic effects

Overall reproductive toxicity

Key result
Reproductive effects observed:
no
Lowest effective dose / conc.:
50 000 ppm
Treatment related:
no

Any other information on results incl. tables

This citation is being presented to cover apparent gaps in the Kieckebusch and Lang (1960) study regarding effects on reproducitve tissues/organs. Table 1 of the Morrissey et al. paper provides the following information regarding benzyl acetate on male reproductive tissues, relative to control:

Species

Terminal Body Weight

Organ Weight

Sperm Parameters

R. cauda

R. epididymis

R. Testis

Absolute

Relative

Absolute

Relative

Absolute

Relative

Motility (%)

Density

Abnormal (%)

Mouse

Decrease

Decrease

Increase

Decrease

Increase

Decrease

Increase

No difference

No difference

No difference

Rat

Decrease

No difference

No difference

No difference

No difference

No difference

No difference

No difference

No difference

No difference

Regarding female reproductive tissue outcomes, benzyl acetate was noted to significantly increase the mean cycle length in the high-dose group of mice.

Applicant's summary and conclusion

Conclusions:
These data supplement missing endpoints from the Kieckebusch and Lang (1960) Key Study, and fill the gap in that study design. In the National Toxicology Program (Morrissey et al., 1988) 13-week feeding benzyl acetate studies in rats and mice, both sperm parameters (epididymis, cauda epididymis, testis weights, and sperm motility, density and percent abnormal sperm) and estrus cyclicity measurements were collected. Dose levels of 2000 to 3000 mg/kg/day did not affect any endpoints of male reproduction. Estrus cycles were lengthened in female mice receiving >3000 mg/kg/day but this only occurred at a dose levels also causing significant decreases in growth (body weights and body weight gain).
Executive summary:

Regulation (EC) No 1907/2006 (REACH), Annex XI 1.5., establishes, that substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group of substances. The similarity may also be based on the likelihood of common breakdown products via physical and biological processes. Given that the grouping of substances and read-across approach can be justified, data gaps may be fulfilled from data of reference substance(s) within the group. This avoids the need to test every substance for every endpoint, which is highly recommended in the context of animal welfare considerations (Directive 2010/63/EU, 2010).

 

For sodium benzoate, the fertility and developmental toxicity endpoints are filled by read across from a group of structurally similar substances.

 

Benzyl acetate, benzyl alcohol, benzaldehyde, and benzoic acid and its sodium and potassium salt were considered as a single category regarding human health by JECFA as they are all rapidly metabolized and excreted via a common pathway within 24 hrs (JECFA 1997).

 

The data presented below if for benzyl acetate using the read across justification described above and is used as read across for sodium benzoate as requested by ECHA in its letter ‘’Submission FP609849-12 – request for dossier update’’ received by the lead registrant on October 03, 2019 and attached in Section 13 of the IUCLID dossier. 

These data supplement missing endpoints from the Kieckebusch and Lang (1960) Key Study, and fill the gap in that study design. In the National Toxicology Program (Morrissey et al., 1988) 13-week feeding benzyl acetate studies in rats and mice, both sperm parameters (epididymis, cauda epididymis, testis weights, and sperm motility, density and percent abnormal sperm) and estrus cyclicity measurements were collected. Dose levels of 2000 to 3000 mg/kg/day did not affect any endpoints of male reproduction. Estrus cycles were lengthened in female mice receiving >3000 mg/kg/day but this only occurred at a dose levels also causing significant decreases in growth (body weights and body weight gain).