Ethyl butyrate

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 chemical was considered to be 500mg/kg bw/day .When female 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

Justification for type of information:

Weight of evidence approach based on the available information from various test chemical.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on two reproductive toxicity studies on rats
1&2.Teratogenic potential toxicity study of test material was performed in rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data avaialble

Specific details on test material used for the study:

- Molecular formula : C6H12O2
- Molecular weight :116.16 g/mol
- Smiles notation : C(=O)(CCC)OCC
- InChl : 1S/C6H12O2/c1356(7)842/h35H2,12H3
- Substance Type: Organic
- Physical State: Liquid

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

No data avaialble

Sex:

female

Details on test animals or test system and environmental conditions:

1.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

IN-LIFE DATES: From: To: No data available
2.TEST ANIMALS
- Source: Anticimex AB, Sollentuna, Sweden
- Age at study initiation: No data available
- Weight at study initiation: 240-330 g
- Fasting period before study: No data available
- Housing: The females were housed individually and had pentachloro-phenol-free wood shavings as bedding material.
- Diet (e.g. ad libitum): R3-pellets, ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimatization period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%): No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: 1.Distilled water 2..Soybean oil

Details on exposure:

1.PREPARATION OF DOSING SOLUTIONS: Test chemical was dissolved in distilled water to give as dose of 0, 100, 500 or 1000 mg/Kg
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

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
- Lot/batch no. (if required): No data
- Purity: No data
2.PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved in soybean oil prior to treatment.
DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Soybean oil
- Concentration in vehicle: 0, 10, 50, 125, 250 or 500 mg/kg bw/day
- Amount of vehicle (if gavage): 5 ml/kg bw
- Lot/batch no. (if required): No data available
- Purity: No data available

Details on mating procedure:

1.- 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

2.- M/F ratio per cage: 1:5
- Length of cohabitation: No data available
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: No data available
- Further matings after two unsuccessful attempts: No data available
- Verification of same strain and source of both sexes: No data available
- Proof of pregnancy: The day sperm was found in the vaginal smear was considered to be day 0 of gestation
- Any other deviations from standard protocol: No data available

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No Data Available

Duration of treatment / exposure:

Study 1
10 days
Study 2.
10 days, i.e. on gestation days 6 through 15

Frequency of treatment:

Daily

Details on study schedule:

- 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:

Study 1
Doses / Concentrations:
0, 100, 500 and 1000 mg/Kg
Basis:
Study 2
0, 10, 50, 125, 250 or 500 mg/kg bw/day

No. of animals per sex per dose:

Study 1
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
Study 2.
Total: 59 female rats
Control: 9 females
10 mg/kg: 10 females
50 mg/kg: 10 females
125 mg/kg: 10 females
250 mg/kg: 10 females
500 mg/kg: 10 females

Control animals:

yes, concurrent vehicle

Details on study design:

No data avaialble

Positive control:

No data avaialble

Parental animals: Observations and examinations:

Study 1.Survival, clinical sign, body weight and body weight change and food consumption were observed.
Study 2.Each female was observed daily throughout the experimental period for physical condition and for signs of toxicity. Body weights were recorded daily from day 0 through day 21 of gestation. Food and water consumption was recorded for each rat at approximately 3-day intervals beginning on day 0. The position and number of fetuses in utero, number of alive and dead fetuses, resorptions and corpora lutea were recorded.

Oestrous cyclicity (parental animals):

Corpora lutea and Resorptions were examined.

Sperm parameters (parental animals):

No data available

Litter observations:

1.Fetuses weighed and sexed were examined.
2.The sex of each fetus and individual fetal weight, gross external anomalies, and placental weights were recorded.

Postmortem examinations (parental animals):

1&2 Organ weight and gross pathology were examined.

Postmortem examinations (offspring):

1.Examined externally for gross abnormalities, visceral, and skeletal malformations and variations, and crown-rump distance were measured.
2.Every third alive fetus of each litter was preserved and later examined under a dissection microscope for soft tissue anomalies after using free hand razor sectioning technique. The remaining fetuses were fixed and stained for examination for skeletal defects.

Statistics:

1.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-significant difference 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.
2. The differences between exposed groups and control groups with respect to numbers of corpora lutea, implantation sites, alive foetuses per litter, pre implantation loss, postimplantation loss as well as mean foetal weight, maternal body weight, placental weight, water and food consumption, was calculated using Student’s t-test. The number of malformations was analyzed using Chi-square test with Yate’s correction.

Reproductive indices:

No data avaialble

Offspring viability indices:

No data avaialble

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

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):

1.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.

2. A total of four dams died during the treatment, one in the 125 mg/kg/day and one in the 500 mg/kg/day group.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

1.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.

2.The mean maternal body weight gain of the treated groups was compared with that of the control group and was not found to differ significantly.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

1.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.

2.The food consumption was similar in control and treated dams.

Food efficiency:

not specified

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

no effects observed

Description (incidence and severity):

2.The water consumption was similar in control and treated dams.

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

1.When treated with 1000 mg/kg bw, slight increase in resorptions were observed as compared to control. But, the difference was not statistically significant. No statistically significant effect was observed on Corpora lutea/dam, Implants/litter, Resorptions/litter and Live fetuses/litter as compared to control.
2.No significant differences were found between the control group and the treated groups as regards corpora lutea and total implantation sites, alive and dead foetuses, resorptions, preimplantation and postimplantation losses, or male/female ratios.

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

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

Remarks on result:

other: No toxic effects were observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

1&2. The mean fetal weights were significantly increased compared to control values in the 50,125 and 250 mg/kg/day groups.

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:

no effects observed

Description (incidence and severity):

2. A low incidence of gastroschisis was noted during gross external examination in the 125 and 500 mg/kg/day groups. This was not significantly different from the controls. Examination of the fetuses for skeletal alterations revealed a slight decrease in the incidence of bipartite centra of the thoracic vertebrae in the 500 mg/kg/day group and a slight increase in the frequency of unossified hyoid cartilage in the 10 and 125 mg/kg/day groups. The skeletal alterations observed seemed not to appear in a systematic manner and are thus not considered to be due to test chemical exposure. At internal examination of the soft tissues in the dissection microscope, no abnormalities were observed in any fetus.

Histopathological findings:

no effects observed

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Moraltity:
No significant effect was observed on Live fetuses/litter s compared to control.

Body weight:
No significant effect was observed on fetal body weight as compared to control.

Gross pathology:
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.

Histopathology:
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.

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.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

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

Remarks on result:

other: No developmental toxic effects were observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Maternal Body Weight And Body Weight Changes^a

	Dose group
Gestational day	0 g/kg (N=20)	0.1 g/kg (N=20)	0.5 g/kg (N=20)	1. 0 g/kg (N=20)
	Body weight (g)
0	216.7 ±22.5	214.8 ± 16.0	213.7 ± 18.3	212.0± 18.5
3	233.3 ±22.4	232.0 ±17.2	230.1 ± 17.8	227.9 ±19.2
6	248.8 ± 23.0	246.1 ±17.8	244.6 ± 20.0	242.9 ± 20.2
9	261.0 ±24.9	256.6 ±18.4	247.3 ±21.2	234.7 ±24.5**
12	279.9 ± 24.3	273.7 ±19.9	261.1 ±23.4*	247.0 ±25.4"
16	306.5 ± 26.0	300.0 ±23.5	288.0 ±27.1	275.3 ± 24.7**
20	368.5 ±28.0	361.3 ±27.7	352.4 ±34.9	338.1 ±26.7*
20^c	295.3 ±23.0	285.6 ±21.1	272.1 ±26.0*	265.1 ±24.3**
	Body weight change (g)
0-6	32.0 ± 3.8	31.3 ± 4.8	30.9 ± 6.8	30.9 ± 6.0
6-9	12.2 ± 4.1	10.6 ± 5.1	2.7 ± 11.3**	-8.1 ± 12.1**
9-12	18.9 ± 4.2	17.1 ± 4.1	13.8 ± 9.4*	13.5 ± 7.9*
12-16	26.6 ± 6.9	26.3 ± 5.6	26.9 ± 8.6	27.3 ± 8.5
16-20	62.0 ± 6.6	61.3 ± 9.0	64.4 ± 10.8	62.8 ± 5.5
0-20	151.8 ± 13.9	146.5 ± 16.4	138.6 ±25.9	126.3 ± 15.5**
6-20^c	46.6 ± 8.5	39.6 ± 9.6	28.8 ±15.0**	22.2 ± 13.3**

^aValues are means ± SD. N = number of dams.

^b From Day 12, W =20; from Day 16, N= 19.

c Body weight corrected for gravid uterine weight.

* Significantly different from control, p≤0.05.

** Significantly different from control, p≤0.01.

Mean food consumption of pregnant rats^a

	Dose group
Gestational day	0 g/kg	0.1 g/kg	0.5 g/kg	1.0 g/kg
0-3	69.2+ 5.9	69.8 ± 5.6	69.2+ 5.7	67.6 ± 5.7
	(21)*	(19)	(22)	(21)
3-6	76.4+ 6.8	72.3 ± 7.8	73.3 ± 6.2	72.5 ± 5.7
	(20)	(18)	(22)	(21)
6-9	76.7 ± 8.1	72.8 ± 7.2	63.7 ± 11.1"	49.3 ±12.3**
	(22)	(19)	(22)	(21)
9-12	80.7 ± 7.5	75.5 ± 8.5	66.9 ± 9.4"	60.5 ± 19.2"
	(21)	(H)	(21)	(18)
12-16	104.3 ± 10.5	101.8 ± 11.9	92.4+ 13.2*	88.8 ± 9.4"
	(22)	(19)	(19)	(19)
16-20	112.7 db 8.5	111.9± 11.6	109.8 ± 14.0	110.0 ± 10.9
	(22)	(20)	(22)	(19)

^aValues are the means ± SD of the amount of diet consumed expressed in grams per rat.

* N is given in parentheses.

* Significantly different from control, p≤ 0.05.

** Significantly different from control, p≤0.01.

Uterine Implantation Data^a

	Dose group
	0 g/kg (N=20)	0.1 g/Kg (N = 20)	0.5 g/kg (N=20)	1.0 g/kg (N =20)
Corpora lutea/dam	16.0 ± 1.6	15.9 ± 2.2	16.1 ± 2.5	16.5 ±2.6
Implants/litter	14.7 ± 1.5	14.7 ± 2.0	14.9 ± 2.1	14.8 ± 1.9
Resorptions/litter	0.6 ±0.7	0.9 ± 1.2	0.6 ± 0.8	1.3± 1.4
Uterine weight (g)	74.9 ± 9.0	75.6 ±12.1	77.8± 13.1	73.0 ±8.8
Live fetuses/litter	7.9 ± 1.9	7.4 ± 1.8	7.0 ± 1.8	6.7 ±2.2
Male	6.2 ±1.6	6.3 ± 1.8	7.2 ± 1.6	6.8 ± 1.9
Female	14.1 ± 1.6	13.7 ± 2.2	14.2 ± 2.0	13.5 ± 1.7
Total	16.0 ± 1.6	15.9 ± 2.2	16.1 ± 2.5	16.5 ±2.6

^aValues are means ± SD. N = number of litters.

Conclusions:

NOAEL was considered to be 500 mg/kg bw/day for the F0 and F1 generation when pregnant female Sprague-Dawley rats were treated with test chemical on gestation days 6 though 15.

Executive summary:

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

Study 1.

In a Teratogenicity study, Sprague-Dawley female rats were treated with test chemical in 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 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 decrase 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 bwas 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 test chemical orally by gavage for 10 days of gestation.

Study 2.

In a teratogenicity study, female Sprague-Dawley rats were exposed to test chemical by oral gavage for 10 days, i.e. on gestation day 6 through 15. As seen by the results, the mean maternal body weight gain and food consumption of the treated groups was compared to control group. The placental weights were significantly reduced for treated animals, and no significant differences were found between the control group and the treated groups as regards to corpora lutea and total implantation sites, alive and dead fetuses, resorptions, preimplantation and postimplantation losses, or male/female ratios. Test chemical exposure resulted in significantly increased mean fetal weights as compared to control values in the 50, 125 and 250 mg/kg/day-exposed animals. A slight decrease in the incidence of bipartite centra of the thoracic vertebrae were noted in the 500 mg/kg/day group and a slight increase in the frequency of unossified hyoid cartilage in the 10 and 125 mg/kg/day groups. However, the skeletal alterations observed seemed not to appear in a systematic manner and is thus not considered to be due to test chemical exposure. Therefore, NOAEL was considered to be 500 mg/kg bw/day for the F0 and F1 generation when pregnant female Sprague-Dawley rats were treated with test chemical on gestation days 6 though 15.

Based on the data available from different studies test chemical did not showedreproductive toxicityat dose concentration 5000mg/kg bw/day by oral route. Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

 

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from authoritative database.

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:

Study 1.

In a teratogenicity study, Sprague-Dawley female rats were treated with test chemical o fOctyl acetate (Cas 112-14-1)in the concentration of 0,100, 500 and 1000 mg/kg/bw/day orally by gavage. Two female rats died in the 1000 mg/kg one each on GDs 10 and 12, respectively. Also, maternal animals in the high-dose group exhibited elevated incidences of alopecia, rales, red nasal discharge and anal-genital staining. Statistically significant and dose-related decrease in mean body weight, and body weight changes were detected in the 1000 mg/kg groups on GDs 9, 12, 16, and 20 were observed at 1000 mg/kg. 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 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 on GD 6-9, 9-12, and 12- 16 intervals were observed at 500 and 1000 mg/kg as compared with control. The uterine implantation parameters including corpora lutea/dam, implants/litter, resorptions/litter, uterine weight or live fetuses/litter showed no statistically significant or dose-dependent differences between control and any of the treated-groups. Although, a slight increase in the number of resorptions was detected in the 1000 mg/kg group as compared to control, the difference was not significant.In the F1 generation, there was no significant or dose-dependent effects on embryo-fetal lethality or fetal growth for any treatment group.No significant differences or dose-related change were observed on crown rump distance of fetuses were observed as compared to control.The number of litters of at least one malformed fetus and the mean percentage of the litter malformed were significantly elevated in the high-dose group only. External observation revealed only two malformations: one in the 1000 mg/kg group (tail absent) and one in the 500 mg/kg group (micrognathia). Visceral examinations revealed two fetuses with abnormalities only in two fetuses at 1000 mg/kg. All groups exhibited comparably low incidences of malformations (dilated ureters/distended renal pelvises), which are anatomical variations previously observed in historical control fetuses. Fetuses with skeletal malformations were observed in the control, low-dose and high-dose groups at low incidence. In addition, different types of vertebral malformations were observed in four fetuses (four litters) from the high-dose group,but, were not statistically significant as compared to control.In conclusion, the test chemicalduring GD 6-15 at the dose level of 500 mg/ml and 1000 mg/kg resulted in significant maternal toxicity as evidenced by reduction in body weight, body weight gain and food consumption of pregnant female rats. However, the administration of the test chemical at maternally toxic doses of 500 and 1000 mg/kg had no adverse effect on sexual function, fertility and fetus development. The administration of the test chemicalslightly increased the incidence of fetal malformations in rats at 1000 mg/kg, which was seriously toxic for mothers. However, no developmental toxicity was observed at the maternally toxic dose of 500 mg/kg or the maternally non-toxic dose of 100 mg/kg. Some developmental effects produced only at maternally toxic exposure levels which indicates that Octyl acetateis not a selective developmental toxicant in rat and it does not disrupt development without obvious signs of adult toxicity. Hence, the NOAELfor reproductive and developmental toxicity is estimated as500 mg/kg/bw/dayfor P0 (parental) generations and 1000 mg/kg/bw/day for F1 generationwhen Sprague-Dawley female rats were orally treated with Octyl acetate for 10 days of gestation under the certain conditions. Maternal toxicity level is 100 mg/kg/bw/day when pregnant female SD rats are orally dosed with the test chemical during gestation.

Study 2.

In a teratogenicity study, female Sprague-Dawley rats were exposed to test chemical of γ-Butyrolactone (96-48-1) by oral gavage at a dose of 0,10,50,125,250 or 500 mg/kg/bw/day for 10 days, i.e. on gestation day 6 through 15). Each female was observed daily throughout the experimental period for physical condition and for signs of toxicity. Body weight, food and water consumption were monitored for each rat at approximately 3-day intervals beginning on day 0. The position and number of fetuses in utero, number of alive and dead fetuses, resorptions and corpora lutea, the sex of each fetus and individual fetal weight, and gross external anomalies and placental weight were recorded.Every third alive fetus were examined for soft tissue anomalies under a dissection microscope, while the remaining fetuses were fixed and stained for examination for skeletal defects. Total of two dams died during the treatment, one in the 125 mg/kg /day and one in the 500 mg/kg/day group due to lung problems.The mean maternal body weight gain and food consumption of all the treated groups was comparable to control group. Although the placental weights were significantly reduced for treated animals, no significant and dose-dependent differences were found between the control and the treated groups regarding the number of corpora lutea and total implantation sites, alive and dead fetuses, resorptions, preimplantation and post-implantation losses, or male/female ratios. Test chemical exposure resulted in significantly increased mean fetal weights as compared to control values in the 50, 125 and 250 mg/kg/day-exposed animals. A slight decrease in the incidence of bipartite centra of the thoracic vertebrae were noted in the 500 mg/kg/day group and a slight increase in the frequency of unossified hyoid cartilage in the 10 and 125 mg/kg/day groups. However, the skeletal alterations observed seemed not to appear in a systematic manner and is thus not considered to be due to test chemical exposure. Therefore, NOAEL was considered to be 500 mg/kg bw/day for the F0 and F1 generation when pregnant female Sprague-Dawley rats were treated with test chemical on gestation days 6- 15.

Based on the data available from different studies test chemical did not showed reproductive toxicity at dose concentration 5000mg/kg bw/day by oral route.Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

 

Effects on developmental toxicity

Description of key information

Developmental toxicity study

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAEL for test chemical was considered to be 500mg /kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

 

Link to relevant study records

Reference

Endpoint:

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

Justification for type of information:

Weight of evidence approach based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Principles of method if other than guideline:

WoE report is based on two developmental toxicity studies on rats.
Teratogenic toxicity study of test material was performed in rats

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Molecular formula : C6H12O2
- Molecular weight :116.16 g/mol
- Smiles notation : C(=O)(CCC)OCC
- InChl : 1S/C6H12O2/c1356(7)842/h35H2,12H3
- Substance Type: Organic
- Physical State: Liquid

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

1.TEST ANIMALS
- Sex: Female
- 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
2.TEST ANIMALS
- Source: Anticimex AB, Sollentuna, Sweden
- Age at study initiation: No data available
- Weight at study initiation: 240-330 g
- Fasting period before study: No data available
- Housing: The females were housed individually and had pentachloro-phenol-free wood shavings as bedding material.
- Diet (e.g. ad libitum): R3-pellets, ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimatization period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%): No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available
IN-LIFE DATES: From: To: No data available

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

water

Remarks:

Distilled

Details on exposure:

1.PREPARATION OF DOSING SOLUTIONS: Test chemical was dissolved in distilled water to give as dose of 0, 100, 500 or 1000 mg/Kg
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

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
- Lot/batch no. (if required): No data
- Purity: No data
2.PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved in soybean oil prior to treatment.
DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Soybean oil
- Concentration in vehicle: 0, 10, 50, 125, 250 or 500 mg/kg bw/day
- Amount of vehicle (if gavage): 5 ml/kg bw
- Lot/batch no. (if required): No data available
- Purity: No data available

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No Data Available

Details on mating procedure:

1.- 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
or by observation of sperm in a vaginal rinse were referred to as day 0 of pregnancy: Copulatory plug in the vagina
- 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 available

2.- M/F ratio per cage: 1:5
- Length of cohabitation: No data available
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: No data available
- Further matings after two unsuccessful attempts: No data available
- Verification of same strain and source of both sexes: No data available
- Proof of pregnancy: The day sperm was found in the vaginal smear was considered to be day 0 of gestation
- Any other deviations from standard protocol: No data available

Duration of treatment / exposure:

1.&2.
10 days

Frequency of treatment:

Daily

Duration of test:

1.
Gestation days-20
2.
21 days

Remarks:

1.
Doses / Concentrations:
0, 100, 500 and 1000 mg/kg bw
Basis:
2.
0, 10, 50, 125, 250 or 500 mg/kg bw/day

No. of animals per sex per dose:

1.
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
2.Total: 59 female rats
Control: 9 females
10 mg/kg: 10 females
50 mg/kg: 10 females
125 mg/kg: 10 females
250 mg/kg: 10 females
500 mg/kg: 10 females

Control animals:

yes, concurrent vehicle

Details on study design:

No data avaialble

Maternal examinations:

1.Survival, clinical sign, body weight and body weight change, food consumption, Organ weight and gross pathology were examined.

2.Each female was observed daily throughout the experimental period for physical condition and for signs of toxicity. Body weights were recorded daily from day 0 through day 21 of gestation. Food and water consumption was recorded for each rat at approximately 3-day intervals beginning on day 0.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes

Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
- Head examinations:No data

Statistics:

1.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.

2.The differences between exposed groups and control groups with respect to numbers of corpora lutea, implantation sites, alive foetuses per litter, preimplantation loss, postimplantation loss as well as mean foetal weight, maternal body weight, placental weight, water and food consumption, was calculated using Student’s t-test. The number of malformations was analyzed using Chi-square test with Yate’s correction.

Indices:

No data avaialble

Historical control data:

No data avaialble

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

2.A total of four dams died during the treatment, one in the 125 mg/kg/day and one in the 500 mg/kg/day group.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The mean maternal body weight gain of the treated groups was compared with that of the control group and was not found to differ significantly.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

The food consumption was similar in control and treated dams.

Food efficiency:

not specified

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

no effects observed

Description (incidence and severity):

The water consumption was similar in control and treated dams.

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

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

Description (incidence and severity):

Placental weights were significantly reduced for treated animals.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

At necropsy, voluminous, oedematous, hyperaemic and emphysematous lungs were seen in the four dams that died during the treatment . There were no signs of aspiration.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Number of abortions:

not specified

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

2.No significant differences were found between the control group and the treated groups as regards to preimplantation and postimplantation losses.

Total litter losses by resorption:

not specified

Early or late resorptions:

no effects observed

Description (incidence and severity):

No significant differences were found between the control group and the treated groups as regards to resorptions.

Dead fetuses:

no effects observed

Description (incidence and severity):

No significant differences were found between the control group and the treated groups as regards to alive and dead foetuses.

Changes in pregnancy duration:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified

Changes in number of pregnant:

not specified

Other effects:

not specified

Description (incidence and severity):

2.No significant differences were found between the control group and the treated groups as regards corpora lutea and total implantation sites, or male/female ratios.

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
Moraltity:
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.

Clinical signs:
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.

Body weight:
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:
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.

Reproductive function: estrous cycle: When treated with 1000 mg/kg bw, slight increas in resorptions were observed as compared to control. But, the difference was not statistically significant.

Reproductive performance: No statistically significant effect were observed on Corpora lutea/dam, Implants/litter, Resorptions/litter and Live fetuses/litter as compared to control.

Organ weights: No statistically significant effect were observed Uterine weight of treated female rats as compared to control.

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Remarks on result:

other: no toxic effects observed

Abnormalities:

not specified

Localisation:

not specified

Description (incidence and severity):

not specified

Fetal body weight changes:

effects observed, treatment-related

Description (incidence and severity):

The mean fetal weights were significantly increased compared to control values in the 50,125 and 250 mg/kg/day groups.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified

Reduction in number of live offspring:

not specified

Changes in sex ratio:

no effects observed

Description (incidence and severity):

No significant differences were observed between the male/female ratios.

Changes in litter size and weights:

not specified

Changes in postnatal survival:

not specified

External malformations:

effects observed, treatment-related

Description (incidence and severity):

1.A low incidence of gastroschisis was noted during gross external examination in the 125 and 500 mg/kg/day groups. This was not significantly different from the controls.
2.A low incidence of gastroschisis was noted during gross external examination in the 125 and 500 mg/kg/day groups. This was not significantly different from the controls.

Skeletal malformations:

effects observed, treatment-related

Description (incidence and severity):

Examination of the fetuses for skeletal alterations revealed a slight decrease in the incidence of bipartite centra of the thoracic vertebrae in the 500 mg/kg/day group and a slight increase in the frequency of unossified hyoid cartilage in the 10 and 125 mg/kg/day groups. The skeletal alterations observed seemed not to appear in a systematic manner and are thus not considered to be due to test chemical exposure.

Visceral malformations:

no effects observed

Description (incidence and severity):

2.At internal examination of the soft tissues in the dissection microscope, no abnormalities were observed in any fetus.

Other effects:

not specified

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Moraltity:
No significant effect was observed on Live fetuses/litter s compared to control.

Body weight:
No significant effect was observed on fetal body weight as compared to control.

Gross pathology:
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.

Histopathology:
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.

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.

Dose descriptor:

NOAEL

Effect level:

> 500 - <= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Remarks on result:

other: No developmental toxic effects observed

Abnormalities:

not specified

Localisation:

other: not specified

Developmental effects observed:

not specified

Treatment related:

not specified

Relation to maternal toxicity:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Conclusions:

NOAEL was considered to be 500 mg/kg bw/day for the F0 and F1 generation when pregnant female Sprague-Dawley rats were treated with test chemical on gestation days 6 though 15.

Executive summary:

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

Study 1:

In a Teratogenicity study, Sprague-Dawley female rats were treated with test chemical in 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 ano-genital staining were observed in treated female rats as compared to control. A statistically significant decreased in mean body weight on 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 decrase 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 bwas 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 test chemical orally by gavage for 10 days of gestation.

Study 2:

In a teratogenicity study, female Sprague-Dawley rats were exposed to test material by oral gavage for 10 days, i.e. on gestation day 6 through 15. As seen by the results, the mean maternal body weight gain and food consumption of the treated groups was compared to control group. The placental weights were significantly reduced for treated animals, and no significant differences were found between the control group and the treated groups as regards to corpora lutea and total implantation sites, alive and dead fetuses, resorptions, preimplantation and postimplantation losses, or male/female ratios. Test chemical exposure resulted in significantly increased mean fetal weights as compared to control values in the 50, 125 and 250 mg/kg/day-exposed animals. A slight decrease in the incidence of bipartite centra of the thoracic vertebrae were noted in the 500 mg/kg/day group and a slight increase in the frequency of unossified hyoid cartilage in the 10 and 125 mg/kg/day groups. However, the skeletal alterations observed seemed not to appear in a systematic manner and is thus not considered to be due to test chemical exposure. Therefore, NOAEL was considered to be 500 mg/kg bw/day for the F0 and F1 generation when pregnant female Sprague-Dawley rats were treated with test material on gestation days 6 though 15.

Thus, Based on the data available fortest chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 500mg /kg bw . Thus, comparing this value with the criteria of CLP regulationtest chemicalis not likely to classify as reproductive and developmental toxicant.

 

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from authoritative database

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Developmental toxicity study

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

Study 1

In a Teratogenicity study, Sprague-Dawley female rats were treated with test chemical in the concentration of0,100, 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 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 decrase 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 was 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 test chemical orally by gavage for 10 days of gestation.

Study 2.

In a teratogenicity study, female Sprague-Dawley rats were exposed to test chemical by oral gavage for 10 days, i.e. on gestation day 6 through 15. As seen by the results, the mean maternal body weight gain and food consumption of the treated groups was compared to control group. The placental weights were significantly reduced for treated animals, and no significant differences were found between the control group and the treated groups as regards to corpora lutea and total implantation sites, alive and dead fetuses, resorptions, preimplantation and postimplantation losses, or male/female ratios. Test chemical exposure resulted in significantly increased mean fetal weights as compared to control values in the 50, 125 and 250 mg/kg/day-exposed animals. A slight decrease in the incidence of bipartite centra of the thoracic vertebrae were noted in the 500 mg/kg/day group and a slight increase in the frequency of unossified hyoid cartilage in the 10 and 125 mg/kg/day groups. However, the skeletal alterations observed seemed not to appear in a systematic manner and is thus not considered to be due to test chemical exposure. Therefore, NOAEL was considered to be 500 mg/kg bw/day for the F0 and F1 generation when pregnant female Sprague-Dawley rats were treated with test chemical on gestation days 6 though 15.

Thus, Based on the data available for test chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 500mg /kg bw . Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

 

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 and developmental toxicant.