3,7-dimethyl-1-octyl acetate

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

Reproductive toxicity study:

Based on the data available from different studies, NOAEL for test material was considered to be 1000 mg/kg /day for reproductive toxicity, when rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data from various test chemicals

Justification for type of information:

Weight of evidence approach based on the data of the read-across chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on 2 reproductive toxicity studies i.e. WoE-2 and WoE-3.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

other: 2. SPF 3. Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

2. - Source: Mollegaards Breeding Centre Ltd., Ejby (DK-4623 L. Skensved).
- Age at study initiation: 4 weeks old
- Weight at study initiation:
- Fasting period before study: not specified
- Housing: The animals were kept in stainless steel wire cages (2 per cage)
- Diet (e.g. ad libitum): The rats were fed a pelleted diet (Chow 101, Institute of Toxicology, and National Food Agency). ad libitum
- Water (e.g. ad libitum): acidified water (citric acid, pH 3.5) was given ad lib.
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23’C ± 1°C,
- Humidity (%): 60 % ± 5%
- Air changes (per hr): air change 6-8 times/h,
- Photoperiod (hrs): electric light from 21.00 to 09.00 hours
3. TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: (P) x wks; 9 weeks old
- Weight at study initiation: 212 - 216 g female
- Fasting period before study: No data available
- Housing: Animals were housed individually (except during the first week of quarantine and during mating) in suspended stainless-steel cages and All animals were identified by uniquely numbered ear tags during the course of the study.
- Diet (e.g. ad libitum): Food (certified Rodent Chow, Ralston Punna Co.)
- Water (e.g. ad libitum): water, ad libitum
- Acclimation period: 3-week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19- 24 °C (monitored daily)
- Humidity (%): 40-70% (monitored daily)
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12-hr photoperiod

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: 2. soya oil 3. Distilled water

Details on exposure:

2. - Justification for use and choice of vehicle (if other than water): Soya oil
- Concentration in vehicle: 0,200,400 and 800 mg/kgbw/ day
3. PREPARATION OF DOSING SOLUTIONS: Test chemical was dissolved in distilled water to give as dose of 0, 100, 500 or 1000 mg/Kg

VEHICLE
- Justification for use and choice of vehicle (if other than water): Distilled water
- Concentration in vehicle: 0, 100, 500 or 1000 mg/Kg
- Amount of vehicle (if gavage): Dose volumes were based on GD 6 body weights throughout the dosing period

Details on mating procedure:

2. - Any other deviations from standard protocol: Histopathology of reproductive organ were examined.
3. - M/F ratio per cage: No data avaialble
- Length of cohabitation: No data avaialble
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Copulatory plug in the vagina
or by observation of sperm in a vaginal rinse were referred to as day 0 of pregnancy
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility. No data avaialble
- Further matings after two unsuccessful attempts: [no / yes (explain)] No data avaialble
- After successful mating each pregnant female was caged (how): Individually
- Any other deviations from standard protocol: No data avaialble

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

2. 28 days
3. 10 days

Frequency of treatment:

2. Daily
3. Daily

Details on study schedule:

2. No data
3. - Rationale for animal assignment (if not random): Females confirmed to have mated were randomly assigned to one of four experimental groups of 22 mated female rats.

Remarks:

2. 0, 200, 400, 800 mg/kg bw/day

Remarks:

3. Doses / Concentrations: 0, 100, 500 and 1000 mg/Kg

No. of animals per sex per dose:

2. Total: 80
0 (vehicle) mg/kg/day: 10 male, 10 female
200 mg/kg/day: 10 male, 10 female
400 mg/kg/day: 10 male, 10 female
800 mg/kg/day: 10 male, 10 female
3. Total: 85
0 mg/kg bw : 22 female
100 mg/kg bw :20 female
500 mg/kg bw :22 female
1000 mg/kg bw :21 female

Control animals:

yes, concurrent vehicle

Details on study design:

2. Not specified
3. No data avaialble

Positive control:

2. Not specified
3. No data avaialble

Parental animals: Observations and examinations:

2. Survival, Clinical sign, Body weight and weight gain and clinical chemistry were examined.
3. Survival, clinical sign, body weight and body weight change and food consumption were observed.

Oestrous cyclicity (parental animals):

2. Not specified
3. Corpora lutea and Resorptions were examined.

Sperm parameters (parental animals):

2. Not specified
3. No data available

Litter observations:

2. Not specified
3. Fetuses weight and sexed were examined.

Postmortem examinations (parental animals):

2. Organ weight and histopathology was examined.
3. Organ weight and gross pathology were examined.

Postmortem examinations (offspring):

2. Not specified
3. Examined externally for gross abnormalities, visceral, and skeletal malformations and variations, and crown-rump distance were measured.

Statistics:

2. Body weights, food and water consumption were compared by means of Student’s t-test. Analysis of variance was performed on haematoiogical parameters, clinical chemical parameters and absolute and relative organ weights. The mean values were compared by the method of Tukey.
3. Maternal body weight and body weight change, food consumption, uterine data (i.e., corpora lutea, implants, resorptions), and malformation data were analyzed statistically using Bartlett's test of homogeneity of variance (Snedecor and Cochran, 1967) was used to determine if the groups had equivalent variances at the 1 % level of significance. If the variances were not significantly different, the groups were compared using a standard one-way analysis of variance (ANOVA). If significant differences among the means were indicated, Duncan's test (Dunnett, 1964) was performed to determine which treated groups differed from control. Fetal weights and crown-rump lengths were analyzed using individual fetal values by a standard nested analysis of variance, with values nested within dams and dams nested within groups. If differences in groups were indicated, the least-significantdifference technique (Snedecor and Cochran, 1967) was used to determine which treated groups differed from control. If the groups did not have equivalent variances at the 1 % level, then a Kruskal-Wallis test (nonparametric) was used to assess differences in group means (Hollander and Wolf, 1973). If the means were different, a rank sum comparison was used to determine which treatment groups differed from control.

Reproductive indices:

2. Not specified
3. No data avaialble

Offspring viability indices:

2. Not specified
3. No data avaialble

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

2. When treated wtih 800 mg/kg bw, after 19 days of dosing female rats showed signs of toxic effects. They had pale mucous membranes and showed signs of pain prevent deaths in this group, the dose was reduced to 400 mg/kg.

3. When treated with 1000 mg/kg bw, elevated incidences of alopecia, rales, red nasal discharge, and anal-genital staining were observed in treated female rats as compared to control.

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

2. When treated wtih 800 mg/kg bw, 4 animals died during the study due to accidental intratracheal dosing.

3. When treated with 1000 mg/kg bw, 2 female rats were died one each on GDs 10 and 12, respectively.
No effect on mortality of 100 and 500 mg/kg bw treated female rats were observed as compared to control.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

2. Decreased in body weight gain was observed in treated male and female rats at 200, 400 and 800 mg/kg bw as compared to control.

3. When treated with 1000 mg/kg bw, A statistically significant decreased in mean body weight on on GDs 9, 12, 16, and 20 were observed as compared with control. The effect was most evident during the first several days of dosing, when these animals actually lost weight (GDs 6-9).
When treated with 500 mg/kg bw, Statistically significant decrase in mean body weight and body weight changes were observed at several time points as compared with control. These decrase in body weight appeared to occur in an ordered response to dose.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

2. When treated wtih 800 mg/kg bw, Decreased in food consumption was observed in treated male rat, and in female rats from 3rd week as compared to control.
When treated with 200 and 400 mg/kg bw, Decreased in food consumption was observed in female rat within the first 2 weeks as compared to control.

3. effects observed, treatment-related
When treated with 500 and 1000 mg/kg bw, dose-related decreases in food consumption for the Day 6-9, 9-12, and 12- 16 intervals were observed as compared with control.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

2. When treated with 800 mg/kg bw, dose-dependent decrease of creatinine concentration and a dose dependent increase of both alkaline phosphatase activity and bilirubin content were observed in treated male and female rats as compared to control.

3. not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

2. Cyst-like spaces appeared scattered in the white matter of the cerebellum at 800 mg/kg bw. The findings resembled the changes found in rats dosed with peppermint oil. No histopathological changes were observed in reproductive organ of testis and ovary of treated rats as compared to control.

3. not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Description (incidence and severity):

3. no effects observed
When treated with 1000 mg/kg bw, slight increase in resorptions were observed as compared to control. But, the difference was not statistically significant.

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Description (incidence and severity):

3. no effects observed
No statistically significant effect were observed on Corpora lutea/dam, Implants/litter, Resorptions/litter and Live fetuses/litter as compared to control.

Dose descriptor:

NOAEL

Effect level:

800 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

histopathology: non-neoplastic

Remarks on result:

other: No effect on histopathology of reproductive organs

Remarks:

2

Dose descriptor:

NOAEL

Effect level:

671 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

histopathology: non-neoplastic

Remarks on result:

other: No effect on histopathology of reproductive organs

Remarks:

2

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

clinical signs

body weight and weight gain

food consumption and compound intake

reproductive function (oestrous cycle)

Remarks on result:

other: No effect on survival, clinical sign, body weight, food consumption, reproductive performance and organ weight

Remarks:

3

Critical effects observed:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Description (incidence and severity):

3. no mortality observed
No significant effect was observed on Live fetuses/litter as compared to control.

Body weight and weight changes:

not specified

Description (incidence and severity):

3. no effects observed
No significant effect was observed on fetal body weight as compared to control.

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Description (incidence and severity):

3. no effects observed
Tail absent were observed in 1000 mg/kg bw and micrognathia in 500 mg/kg bw as compared to control. No significant differences or dose-related change were observed on crown rump distance of fetuses were observed as compared to control.
Skeletal malformations: When treated wtih 1000 mg/kg bw, Different types of vertebral malformations were observed in four fetuses (four litters) in the form of incomplete ossification but, was not statistically significant as compated to control.

Histopathological findings:

not specified

Description (incidence and severity):

3. no effects observed
Visceral examinations: When treated wtih 1000 mg/kg bw, dilated lateral cerebral ventricles in two fetuses were observed which are anatomical variations previously observed in historical control fetuses.

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

gross pathology

histopathology: non-neoplastic

Remarks on result:

other: No effect on body weight, gross pathology and histopathology

Remarks:

3

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Conclusions:

Based on the data available from different studies, the No Observed Adverse Effect Level (NOAEL) for test material was considered to be 1000 mg/kg /day for reproductive toxicity, when rats were treated with test material orally by gavage. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Executive summary:

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical. The studies are as mentioned below:

 

In a repeated dose toxicity study, SPF male and female rats were treated with test chemical in the concentration of 0, 200, 400 and 800 mg/kg bw/day orally by gavage for 28 days. At 800 mg/kg bw, after 19 days of dosing female rats showed signs of toxic effects. They had pale mucous membranes and showed signs of pain prevent deaths in this group, the dose was reduced to 400 mg/kg.4 animals died during the study due to accidental intratracheal dosing. Decreased in body weight gain was observed in treated male and female rats at 200, 400 and 800 mg/kg bw as compared to control. Decreased in food consumption was observed in treated male rat at 800 mg/kg bw, and in female rats from 3rd week at400 mg/kg bw as compared to control. Decreased in food consumption was observed in female rat within the first 2 weeks at 200 and 400mg/kg bw as compared to control. Dose-dependent decrease in creatinine concentration and a dose dependent increase of both alkaline phosphatase activity and bilirubin content were observed in treated male and female rats at 800 mg/kg bw as compared to control. Similarly, statistically significant dose-related increase in relative weight of the spleen, liver and brain of the male rat at 800 mg/kg bw and kidneys, spleen, liver and brain of female rats at 400 mg/kg bw were observed as compare to control. In addition, Cyst-like spaces appeared scattered in the white matter of the cerebellum at 800 mg/kg bw. The findings resembled the changes found in rats dosed with peppermint oil. No histopathological changes were observed in reproductive organ of testis and ovary of treated rats as compared to control. Therefore, NOAEL was considered to be 800 mg/kg bw for male and 671 mg/kg bw for female when SPF male and female rats were treated with test chemical orally by gavage for 28 days.

 

In another teratogenicity study, Sprague-Dawley female rats were treated with the given test chemical at the concentration of 0,100, 500 and 1000 mg/kg bw orally by gavage.2 female rats were died at 1000 mg/kg bw one each on GDs 10 and 12, respectively and elevated incidences of alopecia, rales, red nasal discharge, and anal-genital staining were observed in treated female rats as compared to control. A statistically significant decreased in mean body weight on GDs 9, 12, 16, and 20 were observed at 1000 mg/kg bw. The effect was most evident during the first several days of dosing, when these animals actually lost weight (GDs 6-9).Statistically significant decrease in mean body weight and body weight changes were observed at several time points at 500 mg/kg bw as compared with control. These decrease in body weight appeared to occur in an ordered response to dose. Similarly, Dose-related decreases in food consumption for the Day 6-9, 9-12, and 12- 16 intervals were observed at 500 and 1000 mg/kg bw as compared with control. Slight increase in resorptions was observed as compared to control. But, the difference was not statistically significant. In addition, No statistically significant effect were observed on Corpora lutea/dam, Implants/litter, Resorptions/litter and live fetuses/litter and uterine weight of treated female rats as compared to control in P generation. In F1 generation, No significant effect was observed on fetal body weight as compared to control. Tail absent were observed in 1000 mg/kg bw and micrognathia in 500 mg/kg bw as compared to control. No significant differences or dose-related change were observed on crown rump distance of fetuses were observed as compared to control. Dilated lateral cerebral ventricles in two fetuses were observed at 1000 mg/kg bw which are anatomical variations previously observed in historical control fetuses. Different types of vertebral malformations were observed in four fetuses (four litters) in the form of incomplete ossification at 1000 mg/kg bw. But, were not statistically significant as compared to control. Therefore, NOAEL was considered to be 500 mg/kg bw for P generation and 1000 mg/kg bw for F1 generation when Sprague-Dawley female rats were treated with the given test chemical orally by gavage for 10 days of gestation.

 

Based on the data available from different studies, NOAEL for test material was considered to be 1000 mg/kg/day for reproductive toxicity, when rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from handbook or collection of data.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity study:

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical. The studies are as mentioned below:

 

In a repeated dose toxicity study, SPF male and female rats were treated with test chemical in the concentration of 0, 200, 400 and 800 mg/kg bw/day orally by gavage for 28 days. At 800 mg/kg bw, after 19 days of dosing female rats showed signs of toxic effects. They had pale mucous membranes and showed signs of pain prevent deaths in this group, the dose was reduced to 400 mg/kg.4 animals died during the study due to accidental intratracheal dosing. Decreased in body weight gain was observed in treated male and female rats at 200, 400 and 800 mg/kg bw as compared to control. Decreased in food consumption was observed in treated male rat at 800 mg/kg bw, and in female rats from 3rd week at400 mg/kg bw as compared to control. Decreased in food consumption was observed in female rat within the first 2 weeks at 200 and 400mg/kg bw as compared to control. Dose-dependent decrease in creatinine concentration and a dose dependent increase of both alkaline phosphatase activity and bilirubin content were observed in treated male and female rats at 800 mg/kg bw as compared to control. Similarly, statistically significant dose-related increase in relative weight of the spleen, liver and brain of the male rat at 800 mg/kg bw and kidneys, spleen, liver and brain of female rats at 400 mg/kg bw were observed as compare to control. In addition, Cyst-like spaces appeared scattered in the white matter of the cerebellum at 800 mg/kg bw. The findings resembled the changes found in rats dosed with peppermint oil. No histopathological changes were observed in reproductive organ of testis and ovary of treated rats as compared to control. Therefore, NOAEL was considered to be 800 mg/kg bw for male and 671 mg/kg bw for female when SPF male and female rats were treated with test chemical orally by gavage for 28 days.

 

In another teratogenicity study, Sprague-Dawley female rats were treated with the given test chemical at the concentration of 0,100, 500 and 1000 mg/kg bw orally by gavage.2 female rats were died at 1000 mg/kg bw one each on GDs 10 and 12, respectively and elevated incidences of alopecia, rales, red nasal discharge, and anal-genital staining were observed in treated female rats as compared to control. A statistically significant decreased in mean body weight on GDs 9, 12, 16, and 20 were observed at 1000 mg/kg bw. The effect was most evident during the first several days of dosing, when these animals actually lost weight (GDs 6-9).Statistically significant decrease in mean body weight and body weight changes were observed at several time points at 500 mg/kg bw as compared with control. These decrease in body weight appeared to occur in an ordered response to dose. Similarly, Dose-related decreases in food consumption for the Day 6-9, 9-12, and 12- 16 intervals were observed at 500 and 1000 mg/kg bw as compared with control. Slight increase in resorptions was observed as compared to control. But, the difference was not statistically significant. In addition, No statistically significant effect were observed on Corpora lutea/dam, Implants/litter, Resorptions/litter and live fetuses/litter and uterine weight of treated female rats as compared to control in P generation. In F1 generation, No significant effect was observed on fetal body weight as compared to control. Tail absent were observed in 1000 mg/kg bw and micrognathia in 500 mg/kg bw as compared to control. No significant differences or dose-related change were observed on crown rump distance of fetuses were observed as compared to control. Dilated lateral cerebral ventricles in two fetuses were observed at 1000 mg/kg bw which are anatomical variations previously observed in historical control fetuses. Different types of vertebral malformations were observed in four fetuses (four litters) in the form of incomplete ossification at 1000 mg/kg bw. But, were not statistically significant as compared to control. Therefore, NOAEL was considered to be 500 mg/kg bw for P generation and 1000 mg/kg bw for F1 generation when Sprague-Dawley female rats were treated with the given test chemical orally by gavage for 10 days of gestation.

 

Based on the data available from different studies, NOAEL for test material was considered to be 1000 mg/kg/day for reproductive toxicity, when rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

 

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive toxicant.