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Diss Factsheets

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information
 No data available on acetic anhydride or the read-across substance, acetic acid.
Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Non-human data

No fertility data are available for acetic anhydride. In rat studies, no adverse macroscopic or microscopic findings were observed in the gonads following inhalation of acetic anhydride for 13 weeks (HRC, 1996).

Within an aqueous environment, acetic anhydride is rapidly hydrolysed to acetic acid. Therefore, it is anticipated that acetic anhydride will be rapidly and extensively transformed in vivo to acetic acid and, consequently, it is reasonable to consider systemic exposure to the two substances comparable and systemic toxicity endpoints, such as reproduction and development, would be similar in animals exposed to either substance. However, guideline reproduction studies for acetic acid have also not been reported.

In contrast, local toxicity effects of the two chemicals depend upon the molecular structure of the parent substance and its metabolic/chemical fate within the micro-environment of the site of first contact. The maximum tolerated dose (MTD) for acetic anhydride is associated with local effects (irritation, and in more severe instances resulting in reduced water and food intake and bodyweight depression) and occurs at low concentrations (83.5 mg/m3: highest exposure concentration in rat 90-day inhalation study reported by HRC, 1996). The low MTD may reflect the strongly exothermic aqueous hydrolysis of acetic anhydride which would exacerbate the local effects of the acetic acid produced. Certainly, when considered on a ppm (v/v) basis, acetic anhydride is more potent because, on hydrolysis, it delivers 2 moles of acetic acid per mole of anhydride. A consequence of the low MTD of acetic anhydride is that the extent of systemic exposure to acetic acid will be low when acetic anhydride is administered.

In conclusion, considering systemic exposure to acetate following acetic anhydride exposure, the review of acetic acid for REACH has not uncovered any information to undermine the following statement: “Based on human exposure to orally ingested acetic acid from various foods and the lack of evidence that such exposure is related to fertility problems and developmental deficiencies in humans, neither a new multigeneration study nor any other postnatal evaluation or developmental toxicity study are required” (EU DAR, 2008).

Human data

Although no human data is available for this reproductive endpoint, when considering exposure to acetic anhydride at an atmospheric concentration of 1 ppm (4.18 mg/m3) in the rat 90-day toxicity study, the proposed NOAEC for local irritation effects (4.2 mg/m3) results in an insignificant systemic exposure to acetate at this concentration.For an 8-hour day spent at light work, and assuming 100% absorption of acetic anhydride/acid for a worker, would be 4.2 mg/m3* wRV m3/kg bw * [mw acetic anhydride / acetic acid] = 4.2 * 0.144 * [102.09 / 60.05] = 1.03 mg/kg bw. To put this intake into perspective with the known removal of acetate in humans, it has been shown that about ~0.5 mg/kg bw acetate can be removed each minute via endogenous pathways, such as the citric acid cycle, following administration of acetate in a drink (Smith et al., 2007). Daily administration of 40 mg/kg bw/day may be used as a medicinal product (Johnston & Gaas, 2006), and 25 mg/kg bw /day is estimated as average human (infant) dietary intake (Ishiwata et al., 2002), with peak excursions up to 240 mg/kg bw/day (EU DAR, 2008).

Effects on developmental toxicity

Description of key information

Prenatal developmental toxicity studies (similar to OECD 414) in rat, mouse and rabbit (1974) with acetic acid (5% in cider vinegar) by the oral route. Inhalation range-finding study with acetic anhydride in male rats and pregnant female rats.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP status not known, near guideline study, published in peer reviewed literature, limitations in design and/or reporting but otherwise adequate for assessment.
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
GLP compliance:
not specified
Limit test:
no
Species:
rabbit
Strain:
Dutch
Details on test animals or test system and environmental conditions:
Virgin, adult, Dutch-belted female rabbits were individually housed in mesh bottom cages in temperature and humidity-controlled quarters with free access to food and fresh tap water.
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
The females were dosed with the indicated dosages by oral intubations. The controls were sham treated with the vehicle at a level equivalent to the group receiving the highest test dose. The test material was prepared and doses calculated according to the following table:

Dosage Dose Concentration
(mg/kg) (ml/kg) (mg/ml)
250 1 250
251 - 500 2 125 - 250
501 - 750 3 133 - 250
751 - 1000 4 187 - 250
1001 - 1250 5 200 - 250
1251 - 1500 6 208 - 250
1501 - 1600 6.4 235 - 250
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
On Day 0, each doe was given an injection of 0.4 ml of human chorionic gonadotropin (400 IU) via the marginal ear vein. Three hours later, each doe was inseminated artificially with 0.3 ml of diluted semen from a proven donor buck using approximately 20 x 106 motile sperm.
Duration of treatment / exposure:
Days 6 - 18 of gestation
Frequency of treatment:
Daily
Duration of test:
Days 0-29 of gestation
Dose / conc.:
2.5 mg/kg bw/day (nominal)
Dose / conc.:
16 mg/kg bw/day (nominal)
Dose / conc.:
74.3 mg/kg bw/day (nominal)
Dose / conc.:
345 mg/kg bw/day (nominal)
Dose / conc.:
1 600 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Approximately 12 pregnant females.
Control animals:
yes, sham-exposed
Details on study design:
An additional group of mated females was dosed with 2.5mg/kg 6-aminonicotinamide on day 9 and served as a positive control
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 with particular attention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal
Ovaries and uterine content:
On Day 29 all does were subjected to Caesarean section under surgical anaesthesia, and the numbers of corpora lutea, implantation sites, resorption sites and live and dead foetuses were recorded. The urogenital tract of each animal was examined in detail for normality.
Fetal examinations:
The body weights of the live pups were recorded. all foetuses underwent a detailed gross examination for the presence of external congenital abnormalities. The live foetuses of each litter were then placed in an incubator for 24 hours for the evaluation of neonatal survival. All surviving pups were sacrificed, and all pups 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:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Reduced bodyweight observed in dams treated with 1600 mg/kg/day.
Food consumption and compound intake (if feeding study):
no effects observed
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:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
effects observed, treatment-related
Description (incidence and severity):
Abortions observed in all treated groups at incidences of: 0, 2, 1, 1, 2 for the 0, 16, 74.3, 345 and 1600 mg/kg/day treatment groups.
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
effects observed, treatment-related
Description (incidence and severity):
Reduction in the number of live litters from females treated with 345 and 1600 mg/kg/day.
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Details on maternal toxic effects:
Details on maternal toxic effects:
The administration of up to 1600 mg/kg (body weight) of the test material to pregnant rabbits for 13 consecutive days had no clearly discernible effect on nidation or on maternal survival. Report of reduced body weight in dams at 1600 mg/kg bw/day. Some deaths or abortions occurred in all treated groups and some litter losses were reported at 345 and 1600 mg/kg/day. Maternal effects considered consequence of the bactericidal properties of, orally administered, acetic acid within the gastrointestinal tract of female rabbits.
Dose descriptor:
NOAEL
Effect level:
16 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
dead fetuses
number of abortions
Remarks on result:
other:
Remarks:
Maternal effects considered consequence of the bactericidal properties of, orally administered, acetic acid within the gastrointestinal tract of female rabbits.
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
effects observed, treatment-related
Description (incidence and severity):
Reduction in the number of live offspring observed at 16, 345 and 1600 mg/kg/day.
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
The administration of up to 1600 mg/kg (body weight) of the test material to pregnant rabbits for 13 consecutive days had no clearly discernible effect on fetal 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.
Dose descriptor:
NOAEL
Effect level:
1 600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reduction in number of live offspring
changes in sex ratio
fetal/pup body weight changes
changes in litter size and weights
changes in postnatal survival
external malformations
skeletal malformations
visceral malformations
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
The administration of up to 1600 mg/kg (body weight) to pregnant rabbits for 13 consecutive days had no clearly discernible effect on nidation or on maternal or fetal 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.
Executive summary:

In a developmental toxicity study in rabbits, apple cider vinegar, that contains 5% acetic acid, was administered by gavage at dose levels of 0, 16, 74.3, 345 and 1600 mg/kg/day for 13 consecutive days. The authors reported that administration of up to 1600 mg/kg bodyweight of the test material to dams for 13 days revealed no discernible effect on maternal or foetal survival, or on soft of skeletal tissues.

There was a reduction in pregnancy rate at the high dose and from 74.3 mg/kg/day, a dose-dependent decrease in maternal bodyweight gain in dams. Some deaths or abortions occurred in all treated groups and some litter losses were reported at 345 and 1600 mg/kg/day. Maternal effects were much more noticeable than effects on foetal development. It has been concluded that these findings are a consequence of the bactericidal properties of, orally administered, acetic acid within the gastrointestinal tract of female rabbits (EU, 2008). These effects are considered not to be a direct effect on embryonic implantation and development of acetic acid (EU, 2008). It is likely that this accounts for the apparent increased sensitivity of this species to oral administration of acetic acid.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
5 Jan 1994 - 1 Feb 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant, near guideline study, available as unpublished report. Minor restrictions in design and/or reporting but otherwise adequate for assessment.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
No skeletal or soft tissue examinations
Principles of method if other than guideline:
Preliminary range findings study in 5 male and 5 time-mated females per group, exposed to vapours of acetic anhydride for 6h/day for 5 days/week (males) or from days 6-15 post coitum (females).
GLP compliance:
yes
Species:
rat
Strain:
other: Crl:CD BR
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Manston Road, Margate, Kent
- Age at study initiation: Males - approximately 8 weeks old. Females - 9-11 weeks old.
- Housing: 5 / cage in suspended cages with stainless steel mesh floors. The cages of each group were housed on a rack in a separate ventilated chamber to avoid cross contamination following exposures.
- Diet (e.g. ad libitum): SDS Rat and Mouse No. 1 SQC modified maintenance diet. Available in home cages.
- Water (e.g. ad libitum): Tap water available from polypropylene bottles ad libitum in home cages.
- Acclimation period: Males - 12 days, females - 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24°C
- Humidity (%): 40-70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 h light (08.00-20.00) and 12 h dark/24 h.

IN-LIFE DATES: From: 5 January 1994 To: 1 February 1994.

Route of administration:
inhalation: vapour
Type of inhalation exposure (if applicable):
whole body
Vehicle:
unchanged (no vehicle)
Details on exposure:
pressure of 10mm H2O below ambient.
- Air flow rate: approximately 150L/min.
- Air change rate: Not reported
- Treatment of exhaust air: Drawn through an activated charcoal scrubbing system by an extract fan before being vented to atmosphere.

TEST ATMOSPHERE
- Brief description of analytical method used: Gas chromatography.
- Samples taken from breathing zone: yes

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Acetic anhydride concentration was determined 6 times during each exposure (approximately hourly intervals), using gas chromatography. Samples were also analysed for acetic acid.
The absence of test substance aerosol was confirmed on one occasion during the study using a Royco Particle Monitor.

Details on mating procedure:
- Impregnation procedure: purchased timed pregnant (day 1 post coitum)
Duration of treatment / exposure:
5 days/week for 2 weeks (males)
Days 6-15 post coitum (females)
Frequency of treatment:
6h/day
Duration of test:
2 weeks (except at 400 ppm).
Males were exposed to the high dose concentration on one occasion only, due to toxicity and females were not exposed to this concentration (Table 1)

No. of animals per sex per dose:
5
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: randomly assigned, stratified by bodyweight
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During exposure

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: day 1 of pregnancy and then on days 2, 3, 6, 8, 10, 12, 14, 16, 18 & 20 post coitum.

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

WATER CONSUMPTION : Yes
- Time schedule for examinations: Daily, from day 1 of pregnancy in females.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: Ovaries and uteri


OTHER Litter data and foetal examination: for groups 1 and 2 (killed on day 20 of pregnancy) only. Females killed on day 13 (Groups 3 & 4) were examined in a similar way although embryos were not examined or weighed because they were too immature
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: individual foetal weight was recorded (from which litter weight was calculated). Foetal abnormalities were recorded.
Uteri withiout visible implantations were examine dfor evidence of implantation using a modified Salewski technique.
.
Fetal examinations:
- External examinations: Yes: [all per litter]
- Soft tissue examinations: No
- Skeletal examinations: No
- Head examinations: No

Live young were examined externally and then discarded.
Statistics:
None - in view of small group sizes.
Indices:
Pre and post -implanation losses were calculated as percentages .

Pre-implantation loss was calculated as a percentage from the formula:
(No. of corpora lutea - no. of implantations / No. of corpora lutea ) x 100

Post implantation loss was calculated from the formula:
(No. of implantations - no. of live young / No. of implantations) x 100

Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
At 100 ppm, during the first exposure, females showed halfclosed/closed eyes, licking of the inside of the mouth and lachrymation. After further exposures, gasping and/or exaggerated respiration and occasional twitching of the head was seen.
After exposure, noisy respiration, occasional sneezing and gasping respiration, arching of the back and occasional twitching of the head were seen in all /most animals. Severe weight loss was seen at this exposure level along with noticeably lower food and water intake values. Consequently , this group of animals was terminated on day 13 after 7 exposures.

At 25 ppm, half closed/closed eyes were observed during each exposure. Licking of the inside of the mouth and occasional head twitching were also observed predominantly on the last 3-4 days of exposure.
Noisy respiration was noted post exposure and persisted until the day prior to sacrifice (4 days after completion of the last exposure). Bodyweight gain was depressed from day 2 onwards along with food and water intake, compard to controls, but all showed some recovery on cessation of exposures.
Dose descriptor:
LOAEC
Effect level:
24 ppm (analytical)
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
At 100 ppm, females were killed on day 13 and were compared with the unexposed Group 4 females (also killed on day 13 post coitum). Four out of five females were pregnant in each group. Two females exposed to 100 ppm showed total resorption of their litters, compared to none in the control group. The other two pregnant females in this group were supporting live litters of comparable size with those of the unexposed concurrent control group. Examination of foetuses for visual abnormalities was not possible given the age at sacrifice.
At 25 ppm, all females had a live litter at scheduled termination (day 20). There were no differences from controls in the incidence and pattern of embryofoetal loss, litter size, litter weight or foetal weight and there were no gross foetal abnormalities.
Dose descriptor:
NOAEC
Effect level:
24 ppm (analytical)
Basis for effect level:
other: embryotoxicity
Abnormalities:
not specified
Developmental effects observed:
not specified

Table 2 - Litter data

Category

Group and Exposure (analysed concentration – ppm)

1 - Control

2 - 24

3 - 104

4 - Untreated

No. with live young

5

5

2 

(4)

4

Corpora lutea/dam

14.6

14.6

15

(14.5)

15.3

Implantations/dam

14.0

14.4

14.5

(14.3)

14.5

Pre-implantation loss %

3.8

1.3

3.0

(1.5)

4.4

Early embryonic death

1.4

0.6

1.0

(7.5)

2.0

Late embryonic death

0.0

0.0

-

-

-

Total embryonic death

1.4

0.6

-

-

-

Post-implantation loss %

9.9

4.2

7.7

(53.9)

12.0

Live young/dam

12.6

13.8

13.5

(6.8)

12.5

Litter weight

50.31

52.09

NR

NR

Mean foetal weight

3.99

3.78

NR

NR

EMBRYONIC DEATHS

Total (early)                         0

-

2

1

(-)

2

                                              1

3

3

-

(-)

1

                                              2

2

-

1

(-)

-

                                              3

-

-

-

(-)

-

                                        = > 4

 - - -

(2)

 1

NR not recordable

( ) data from all pregnant animals, inclusive of 2 showing total resorption

Executive summary:

In a 2 week preliminary inhalation study, exposure of time-mated females to 100 or 25 ppm acetic anhydride caused clear signs of maternal toxicity (noisy respiration, severely reduced food and water consumption and a marked bodyweight loss). At 100 ppm, females were removed from the study on day 13 after 7 exposures. Two females in this group showed total resorption, probably as a consequence of the marked maternal toxicity.

At 25 ppm, there was clear evidence of maternal toxicity similar to, but less severe than, at higher exposure levels. However, there was no evidence of embryotoxicity at this exposure level.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP status not known, near guideline study, published in peer reviewed literature, limitations in design and/or reporting but otherwise adequate for assessment.
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
Virgin adult female albino rats (Wistar derived stock) were individually housed in mesh bottom cages in temperature and humidity controlled quarters with free access to food and fresh tap water.
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
The females were dosed with the indicated dosages by oral intubations. The controls were sham treated with the vehicle at a level equivalent to the group receiving the highest test dose. The test material was prepared and doses calculated according to the following table:

Dosage Dose Concentration
(mg/kg) (ml/kg) (mg/ml)
250 1 250
251 - 540 2 125 - 250
501 - 750 3 133 - 250
751 - 1000 4 187 - 250
1001 - 1250 5 200 - 250
1251 - 1500 6 208 - 250
1501 - 1600 6.4 235 - 250

Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
The females were mated with young adult males, and observation of the vaginal sperm plug was considered Day 0 of gestation.
Duration of treatment / exposure:
Days 6 - 15 of gestation
Frequency of treatment:
Daily
Duration of test:
Days 0-20 of gestation
Dose / conc.:
16 mg/kg bw/day (nominal)
Dose / conc.:
74.3 mg/kg bw/day (nominal)
Dose / conc.:
345 mg/kg bw/day (nominal)
Dose / conc.:
1 600 mg/kg bw/day (nominal)
No. of animals per sex per dose:
25 mated females
Control animals:
yes, sham-exposed
Details on study design:
An additional group of mated females was dosed with 150 mg/kg aspirin and served as a positive control
Maternal examinations:
Body weights were recorded on Days 0, 6, 11, 15, and 20 of gestation.
All animals were observed daily for appearance and behaviour with particular attention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal
Ovaries and uterine content:
On Day 20 all dams were subjected to Caesarean section under surgical anesthesia, and the numbers of implantation sites, resorption sites, and live and dead fetuses were recorded. The urogenital tract of each dam was examined in detail for anatomical normality.
Fetal examinations:
The body weights of the live pups were recorded. All fetuses were examined grossly for the presence of external congenital abnormalities. One-third of the fetuses of each litter underwent detailed visceral examinations employing the Wilson technique. The remaining two-thirds were cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
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:
no effects observed
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:
no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
not examined
Details on maternal toxic effects:
Details on maternal toxic effects:
The administration of up to 1600 mg/kg (body weight) of the test material to pregnant rats for 10 consecutive days had no clearly discernible effect on nidation or on maternal survival.
Dose descriptor:
NOAEL
Effect level:
1 600 mg/kg bw/day
Basis for effect level:
body weight and weight gain
changes in number of pregnant
changes in pregnancy duration
dead fetuses
early or late resorptions
effects on pregnancy duration
gross pathology
histopathology: non-neoplastic
mortality
necropsy findings
number of abortions
organ weights and organ / body weight ratios
pre and post implantation loss
total litter losses by resorption
Fetal body weight changes:
no effects observed
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:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
The administration of up to 1600 mg/kg (body weight) of the test material to pregnant rats for 10 consecutive days had no clearly discernible effect on fetal 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.
Dose descriptor:
NOAEL
Effect level:
1 600 mg/kg bw/day (nominal)
Basis for effect level:
reduction in number of live offspring
changes in sex ratio
fetal/pup body weight changes
changes in litter size and weights
changes in postnatal survival
external malformations
skeletal malformations
visceral malformations
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
The administration of up to 1600 mg/kg (body weight) to pregnant rats for 10 consecutive days had no clearly discernible effect on nidation or on maternal or fetal 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.
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP status not known, near guideline study, published in peer reviewed literature, limitations in design and/or reporting but otherwise adequate for assessment.
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
GLP compliance:
not specified
Limit test:
no
Species:
mouse
Strain:
CD-1
Details on test animals or test system and environmental conditions:
Virgin adult female albino CD-1 outbred mice were gang-housed in disposable plastic cages in temperature and humidity-controlled quarters with free access to food and fresh tap water.
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
Dose volume 10 ml/kg body weight
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
The females were mated with young adult males, and observation of the vaginal sperm plug was considered Day 0 of gestation.
Duration of treatment / exposure:
Days 6 - 15 of gestation
Frequency of treatment:
Daily
Duration of test:
Days 0-17 of gestation
Dose / conc.:
16 mg/kg bw/day (nominal)
Dose / conc.:
74.3 mg/kg bw/day (nominal)
Dose / conc.:
345 mg/kg bw/day (nominal)
Dose / conc.:
1 600 mg/kg bw/day (nominal)
No. of animals per sex per dose:
25 mated females
Control animals:
yes, sham-exposed
Details on study design:
An additional group of mated females was dosed with 150 mg/kg aspirin and served as a positive control
Maternal examinations:
Body weights were recorded on Days 0, 6, 11, 15, and 17 of gestation.
All animals were observed daily for appearance and behaviour with particular attention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal
Ovaries and uterine content:
On Day 17 all dams were subjected to Caesarean section under surgical anesthesia, and the numbers of implantation sites, resorption sites, and live and dead fetuses were recorded. The urogenital tract of each dam was examined in detail for anatomical normality.
Fetal examinations:
The body weights of the live pups were recorded. All fetuses were examined grossly for the presence of external congenital abnormalities. One-third of the fetuses of each litter underwent detailed visceral examinations employing the Wilson technique. The remaining two-thirds were cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Reduced bodyweight gain observed in females treated with 345 and 1600 mg/kg/day.
Food consumption and compound intake (if feeding study):
no effects observed
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:
no effects observed
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:
effects observed, treatment-related
Description (incidence and severity):
There was an increase in the number of dead foetuses in females treated with 1600 mg/kg/day.
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Details on maternal toxic effects:
Details on maternal toxic effects:
The administration of up to 1600 mg/kg (body weight) of the test material to pregnant mice for 10 consecutive days had no clearly discernible effect on maternal survival.
Dose descriptor:
NOAEL
Effect level:
74.3 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
dead fetuses
Remarks on result:
other: Slight toxicity (reduced bodyweight gain) at 345 and 1600 mg/kg/day
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
effects observed, treatment-related
Description (incidence and severity):
There was a decreased number of live offspring from females treated with 1600 mg/kg/day.
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
effects observed, treatment-related
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
The administration of up to 1600 mg/kg (body weight) of the test material to pregnant mice for 10 consecutive days had no clearly discernible effect on fetal 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.
Dose descriptor:
NOAEL
Effect level:
345 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reduction in number of live offspring
other: Delyaed ossification
Remarks on result:
other: At the highest dose tested, 1600 mg/kg/day, there was an increase in the number of litters containing a dead foetus and some reductions in ossification.
Key result
Developmental effects observed:
yes
Lowest effective dose / conc.:
1 600 mg/kg bw/day (nominal)
Treatment related:
yes
Relation to maternal toxicity:
developmental effects as a secondary non-specific consequence of maternal toxicity effects
Dose response relationship:
not specified
Conclusions:
The NOAEL for maternal toxicity was determined to be 74.3 mg/kg/day, due to reduced bodyweight gain and number of dead foetuses. The NOAEL for developmental toxicity was 345 mg/kg/day, due to number of dead foetuses and delayed ossification.
Executive summary:

In a developmental toxicity study in mice, apple cider vinegar, that contains 5% acetic acid, was administered by gavage at dose levels of 0, 16, 74.3, 345 and 1600 mg/kg/day for 10m consecutive days. The authors reported that administration of up to 1600 mg/kg bodyweight of the test material to dams for 10 days revealed no discernible effect on maternal or foetal survival, or on soft or skeletal tissues.

There was no effect on the foetal development, in the presence of slight maternal toxicity (reduced bodyweight gain) at 345 mg/kg/day. At the highest dose tested, 1600 mg/kg/day, there was an increase in the number of litters containing a dead foetus and some reductions in ossification. The NOAEL for maternal and developmental toxicity in mice were 74.3 mg/kg/day and 345 mg/kg/day, respectively. 

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
345 mg/kg bw/day
Study duration:
subacute
Species:
mouse
Quality of whole database:
NOTE: NOAEL is for developmental effects with apple cider vinegar (5% acetic acid). There are studies in rats, mice and rabbits considered equivalent or similar to EU Method B.31 (Prenatal Developmental Toxicity Study). These all pre-date GLP and are evaluated at Klimisch score=2.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEC
104 mg/m³
Study duration:
subacute
Species:
rat
Quality of whole database:
The key study is GLP compliant. It is a preliminary study and the number of animals are insufficient for a full evaluation of developmental effects however it is reliable (Klimisch=1).
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Non-human data

A range-finding study in which rats were exposed to acetic anhydride by inhalation reported irritation of the eyes and upper respiratory tract of the dams at 25 ppm (104 mg/m3), but no effects on the foetus (HRC, 1994). Increased resorptions, most likely a consequence of maternal toxicity, were reported at 100 ppm (418 mg/m3). At this dose level, over the exposure period, dams displayed continually worsening effects of eye and respiratory tract irritation and, most likely as a consequence, a decrease in food and water consumption, finally expressed as a large decrease in bodyweight. This test group was discontinued on day 13 post-coitum after only 7 exposures. Maternal toxicity following inhalation exposure occurs at relatively low concentrations (100 mg/m3). The MTD may reflect the strongly exothermic aqueous hydrolysis of acetic anhydride that may exacerbate local effects of the acetic acid produced.

Within an aqueous environment, acetic anhydride is rapidly hydrolysed to acetic acid. Developmental toxicity data are available for apple cider vinegar (containing 5% acetic acid). As acetic anhydride is rapidly and extensively transformedin vivoto acetic acid, it is reasonable to consider systemic exposure to the two substances comparable and systemic toxicity endpoints, such as reproduction and development, would be similar in animals exposed to either substance. In contrast, local toxicity effects of the two chemicals are likely to depend upon the molecular structure of the parent substance and its metabolic/chemical fate within the micro-environment of the site of first contact.

Considering the acetate metabolite, developmental toxicity studies in rats, mice and rabbits have been reported for apple cider vinegar, that contains 5% acetic acid, administered by gavage at dose levels of 0, 16, 74.3, 345 and 1600 mg/kg/day (Morgareidge, 1974). There was no evidence of a direct embryo toxicity or teratogenic potential and the NOAEL values for maternal and developmental toxicity in mice were 74 and 345 mg/kg/day, respectively.

The low systemic exposure to acetate, following administration of acetic anhydride at concentrations below the NOAEC for local effects is discussed for reproductive toxicity. Such considerations are also relevant to developmental toxicity.

Human data

No relevant human information is available.

Justification for classification or non-classification

There is no evidence to indicate adverse reproductive or developmental potential to humans for acetic anhydride. Therefore classification is not warranted.

Additional information