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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
2008

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified
Details on test material:
- Name of test material (as cited in study report): diallyl phthalate (identity verified by GC/MS)
- Supplier: Tokyo Chemical Industry, Tokyo, Japan
- Analytical purity: > 98% (confirmed by GC)
- Stability under test conditions: for up to 14 days (when stored in a dark place, at room temperature)
- Storage condition of test material: in a dark place, at room temperature

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: IFFA CREDO Breeding Laboratories (St-Germain-sur-l'Arbresle, France)
- Acclimation period: 1-2 weeks
- Weight at study initiation: 180-200 g
- Housing: single housing in clear polycarbonate cages with stainless steel wire lids and corn cob granules as bedding
- Diet: food pellets (UAR Alimentation Villemoisson, France), ad libitum
- Water (e.g. ad libitum): filtered tap water ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21±2
- Humidity (%): 50±5
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis by gas chromatography showed that the formulations used for dosing were stable for up to 14 days (storage in the dark at room temperature).
Details on mating procedure:
- Impregnation procedure: cohoused with M/F ratio per cage = 1/1.
- Further matings after unsuccessful attempts: not specified but more than 90% of the matings were successful in all groups.
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy and in all groups more than 90% of the females became pregnant.
Duration of treatment / exposure:
Dosing occurred once daily, in the morning, on day 6 to day 20
Frequency of treatment:
Dosing occurred once daily, in the morning, on day 6 to day 20
Duration of test:
From mating to GD 21
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Remarks:
Control (olive oil)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
150 mg/kg bw/day (actual dose received)
Dose / conc.:
200 mg/kg bw/day (actual dose received)
Dose / conc.:
250 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
Group size ranged from 23 or 24 time-mated rats (20-23 pregnant).
Control animals:
yes, concurrent vehicle

Examinations

Maternal examinations:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: GD 0, 6, 9, 12, 15, 18 and 21

FOOD CONSUMPTION: Yes
- Food consumption: at three-day intervals starting on GD 6 for each individual female expressed as mean daily diet consumption in g food/rat/day

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined: uterus
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: Number of dead and live fetuses
Fetal examinations:
- External examinations: Yes [all per litter]
- Soft tissue examinations: Yes [half per litter]
- Skeletal examinations: Yes [half per litter]
Statistics:
- All data presented as mean ± SD where possible;
- Number of corpora lutea, implantation sites, live fetuses, body weights: one-by-one analysis of variance, followed by Dunnett's test if differences were found.
- Frequency of post-implantation loss, dead fetuses, resoprtions, alterations among litters: Kruskal-Wallis test followed by the Mann-Whitney test
- Rates of pregnancy and of litters with dead fetuses or resoprtions, incidences of fetal alterations per dose: Fisher's test
Historical control data:
Not specified.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Maternal weight gain and food consumption were significantly reduced at 200 and 250 mg/kg/day.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Maternal weight gain and food consumption were significantly reduced at 200 and 250 mg/kg/day.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Immunological findings:
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Maternal weight gain and food consumption were significantly reduced at 200 and 250 mg/kg/day.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Gross examination at necropsy revealed liver lesions in dams given 150 mg/kg/day or higher doses.
Neuropathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Fetal body weight was significantly reduced at 200 and 250 mg/kg/day. There was a significant increase in the incidence of fetuses with skeletal variations at 250 mg/kg/day. Retarded ossification of certain bones (i.e. fore-limb and hind limb phalanges, metatarsals, and caudal vertebrae) was also observed.
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
200 and 250 mg/kg/d: dose-related effects: decrease in food consumption and body weight gain
250 mg/kg/d: 1 dead
150 mg/kg/d: macroscopic changes in the liver

Effect levels (maternal animals)

open allclose all
Key result
Dose descriptor:
NOEL
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Key result
Dose descriptor:
NOEL
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Dose descriptor:
NOEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Fetal body weight was significantly reduced at 200 and 250 mg/kg/day.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): effects observed, treatment-related
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Fetal body weight was significantly reduced at 200 and 250 mg/kg/day.
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
effects observed, treatment-related
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
There was a significant increase in the incidence of fetuses with skeletal variations at 250 mg/kg/day. Retarded ossification of certain bones (i.e. fore-limb and hind limb phalanges, metatarsals, and caudal vertebrae) was also observed.
Visceral malformations:
no effects observed
Other effects:
not examined
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
100 to 250 mg/kg/d: dose related decrease in fetal body weight significant at and above 200 mg/kg/d (see attached graph).
100 to 250 mg/kg/d: exponential increase of skeletal variations with increasing dose (see attached graph) and incomplete skeletal ossification of some bones (forelimb and hindlimb phalanges, metatarsals)

Effect levels (fetuses)

Key result
Dose descriptor:
LOAEL
Effect level:
250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
skeletal malformations

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Applicant's summary and conclusion

Conclusions:
The present study demonstrated that oral administration of DAP to rats during the time of implantation to the term of pregnancy produced significant foetotoxicity at doses of 200 mg/kg/d or higher, which were also maternally toxic. However, no significant embryolethality or teratogenicity was recorded at any of the doses tested.
Executive summary:

The objective of this study was to evaluate the developmental toxic potential of diallyl phthalate (DAP) in rats. Pregnant Sprague-Dawley rats were given DAP at doses of 0 (olive oil), 100, 150, 200, and 250 mg/kg/day, by gavage (5 ml/kg), on Gestational Days (GD) 6 through 20. Gross examination at necropsy revealed liver lesions in dams given 150 mg/kg/day or higher doses. In addition, maternal weight gain and food consumption were significantly reduced at 200 and 250 mg/kg/day. There was no significant increase in the incidence of resorptions, or malformations, at any dose. Fetal body weight was significantly reduced at 200 and 250 mg/kg/day. There was a significant increase in the incidence of fetuses with skeletal variations at 250 mg/kg/day. Retarded ossification of certain bones (i.e. fore-limb and hindlimb phalanges, metatarsals, and caudal vertebrae) was also observed. There was no sign of developmental toxicity at 100 and 150 mg/kg/day. Thus, DAP caused fetal toxicity at doses which also produced maternal effects, but no embryolethality or teratogenicity.