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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Conducted according to sound scientific principles, similar to OECD TG 421. GLP.

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Unnamed
Year:
1998
Reference Type:
secondary source
Title:
Tris(2-ethylhexyl)benzene-1,2,4-tricarboxylate
Author:
ACC Phthalate Esters Panel HPV Testing Group
Year:
2006
Bibliographic source:
IUCLID 4 Data Set, HPV Chemical

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
other: OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Tris(2-ethylhexyl)benzene-1,2,4-tricarboxylate
Source: Daihachi Kagaku Kogyo Co., Ltd. Lot. No. N-80301
Purity: >99.0% Kept at room temperature in a dark place until use.

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Details on test animals and environmental conditions:
Age at study initiation: 10 week old for both sexes.
Weight at study initiation: 373-435 g for males, 217-257 g for females
No. of animals per sex per dose: 12 per sex per dose group

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
Administration period: Males for 46 days from 2 weeks prior to mating; Females from 2 weeks prior to mating to day 3 of lactation
Post exposure observation period: None
Terminal kill: Males day 47; Females day 4 of lactation
Analytical verification of doses or concentrations:
no
Details on mating procedure:
Male/female per cage; 1/1, length of cohabitation; with in the limit of 14 days until proof of pregnancy (formation sperm detection in vagina) was observed.
Duration of treatment / exposure:
Administration period: Males for 46 days from 2 weeks prior to mating;
Females from 2 weeks prior to mating to day 3 of lactation
Frequency of treatment:
Once a day
Duration of test:
Administration period: Males for 46 days from 2 weeks prior to mating;
Females from 2 weeks prior to mating to day 3 of lactation
No. of animals per sex per dose:
12 per sex per dose group
Control animals:
yes, concurrent vehicle
Details on study design:
Administration period: Males for 46 days from 2 weeks prior to mating;
Females from 2 weeks prior to mating to day 3 of lactation
Post exposure observation period: None
Terminal kill: Males day 47; Females day 4 of lactation

The animals were sacrificed on the day 4 of lactation for females.
Males and females with no mated were killed 1 day after the mating period.
Females with no delivery killed 26th day of gestation period.
Vehicle: Corn oil
Mating procedures: Male/female per cage; 1/1, length of cohabitation; with in the limit of 14 days until proof of pregnancy (formation sperm detection in vagina) was observed.

Examinations

Maternal examinations:
Clinical observations performed and frequency:
Parent: General appearance once a day

Parameters assessed during study:
Body weight.
Males: Prior to the first dosing and 2, 5, 7, 10, 14 day. After that once a week, the day sacrificed.
Females: Prior to the first dosing and 2, 5, 7, 10, 14 day. During gestation period, 0, 1, 3, 5, 7, 10, 17 and 20 day.
During lactation period, 0, 1, and 4.
During cohabitation period, the same day with male.

Food/water consumption.
The same day when body wt. determined, except lactation period and the day sacrificed for males, also, 0 day of gestationand lactation for female.
Ovaries and uterine content:
Organs examined at necropsy:
Parent: Males and females: Gross pathlogy of all organs were tested.
Males: Organ weight: Testis and epididymis of all animals.
Female: Organ weight: Ovary of all animals.

Count: Implantation sites and corpus luteum of ovary of all animals.

Microscopic:
Males: Testis and epididymis. Count of sertoli sells, spermatocytes, round spermatids and elongate spermatids in seminiferous tubules of 5 animals of all dosing groups.(Stage I-VI, VII-VIII, IX-XI, XII-XIV of spermatozoon formative cycle.)
Females: Ovary
Fetal examinations:
Clinical observations performed and frequency:
Pups: General appearance once a day after birth

Parameters assessed during study:
Body weight.
Pups: Day 0 and 4

Postmortem: Gross pathlogy of all organs were tested. Dead pups and abnormal organs were tested histopathogy.
Statistics:
Chi square test for 1 grade positive data and Fisher's test for another. Bartlett's test or Kruskal-Wallis' test for 2 or more grade positive data. And used Dunnett's test or Mann-Whitney's U-test for examination.
Indices:
Reproductive indices
No. of pairs with successful copulation;
Copulation index (No. of pairs with successful copulation/No. of pairs mated) x 100, duration of mating;
No. of pregnant females;
Fertility index = (No. of pregnant animals/No. of pairs with successful copulation) x 100

Offspring viability indices
No. of corpora lutea;
No. of implantation sites;
Implantation index (No. of implantation sites/No. of corpora lutea) x 100;
No. of pups born, delivery index (No. of pups born/No. of implantation sates) x 100;
No. of live pups born;
Live birth index (No. of live pups born/No. of pups born) x 100, sex ratio of pups;
No. of dead pups born;
Gestation length;
Gestation index (No. of females with live pops delivered/ No. of pregnant females) x 100;
Nursing index (No. of females nursing live pups/No. of females with normal delivery) x 100;
No. of live pups on day 4, viability index (No. of live pups on day 4/No. of live pups born) x 100.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
Mortality and day of death: None

Body weight: No statistical significant difference from controls.

Food/water consumption: No statistical significant difference from controls.

Reproductive data: No statistical significant difference from controls.

Grossly visible abnormalities, external, soft tissue and skeletal abnormalities (females):
Cyst of corpus luteum of ovary was observed 2 animals of 300 mg/kg dosing group. No abnormal ovaries were observed in the control females 100 mg/kg dosing without successful copulation, or in the 100 mg/kg dosing group without pregnant.

There were no effects observed in any of the following parameters:
1) No. of pairs with successful copulation;
2) Duration of mating ;
3) Copulation index(%);
4) No. of pregnant animals,
5) Fertility index(%);
6) No. of corpora lutea;
7) No. of implantation sites;
8) Implantation index(%);
9) No. of pups born(%);
10) Delivery index(%);
11) Live pups born;
12) Live birth index(%);
13) Sex ratio(M/F);
14) Dead pups born;
15) Gestation length(day);
16) Gestation index(%);
17) Nursing index(%)

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Body weight and weight gain of 300 mg/kg dosing group for both sexes were slightly low. But all pups of 100 and 1000 mg/kg dosing group were not statistical significant difference from controls. At the other tests, no statistical significant difference from controls. No gross pathological changes were observed.

1) No. of pups born (%) – no effects observed;
2) Delivery index (%) – no effects observed;
3) Live pups born – no effects observed;
4) Live birth index(%) – no effects observed;
5) Sex ratio (M/F) – no effects observed;
6) Dead pups born – no effects observed;
7) Live pups on day 4 – no effects observed;
8) Viability index (%)– no effects observed;
9) Body weight of pups – only the 300 mg/kg dosing group for both sexes were statistically significantly decreased;
10) Day 0-4, weight-gain (M and F, in g) – only the 300 mg/kg dosing group for both sexes were statistically significantly decreased;
11) Body weight gain (%) (M and F) – no effects observed..

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: fetotoxicity

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The reproductive NOAEL for the test material was determined to be >1000 mg/kg bw/day, the highest dose tested, for male and female rats (P generation). The developmental NOAEL was determined to be >1000 mg/kg bw/day, the highest dose tested, for the F1 generation as no fetotoxic, developmental, or teratogenic effects were observed.

Classification as a reproductive or developmental toxicant is not warranted under the new Regulation (EC) 1272/2008 on classification, labeling and packaging of substances and mixtures (CLP) or under Directive 67/548/EEC for dangerous substances and Directive 1999/45/EC for preparations.
Executive summary:

Male Crj: CD(SD) rats were treated with the test material for 46 days starting from 2 weeks prior to mating; female Crj: CD(SD) rats were treated with the test material starting from 2 weeks prior to mating to day 3 of lactation. Male amd female rats were cohabitated for up to 14 days or until proof of pregnancy was observed. Clinical observations, body weight, and food/water consumption were observed during the experimental period. Several indices for reproductive and for developmental performance were evaluated. Both male and female rats as well as the resulting pups were sacrificed and examined for gross pathology and for histopathological changes to the reproductive organs.

 

The reproductive NOAEL for the test material was determined to be >1000 mg/kg bw/day, the highest dose tested, for male and female rats (P generation). The developmental NOAEL was determined to be >1000 mg/kg bw/day, the highest dose tested, for the F1 generation as no fetotoxic, developmental, or teratogenic effects were observed.

 

Classification as a reproductive or developmental toxicant is not warranted under the new Regulation (EC) 1272/2008 on classification, labeling and packaging of substances and mixtures (CLP) or under Directive 67/548/EEC for dangerous substances and Directive 1999/45/EC for preparations.