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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Justification for type of information:
ANALOGUE APPROACH JUSTIFICATION
Please refer to attached document.

Data source

Reference
Reference Type:
review article or handbook
Title:
Unnamed
Year:
2007
Report date:
2007

Materials and methods

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

Test material

Constituent 1
Chemical structure
Reference substance name:
Diallyl phthalate
EC Number:
205-016-3
EC Name:
Diallyl phthalate
Cas Number:
131-17-9
Molecular formula:
C14H14O4
IUPAC Name:
1,2-Benzenedicarboxylic acid, di-2-propenyl ester
Test material form:
liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Commercial laboratory animal supplier
- Age at study initiation:
- Weight at study initiation:
- Fasting period before study:
- Housing: clear polycarbonate cages with stainless steel wire lids and corn cob granules for bedding.
- Diet; food pellets ad libitum
- Water; ad libitum
- Acclimation period: 1-2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2°C
- Humidity (%): 50 ± 5%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12h light-dark photocycle

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food: ambient

Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: no data
- Length of cohabitation:
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- Any other deviations from standard protocol:
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
150 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
No. of animals per sex per dose:
12
Control animals:
yes
Details on study design:
- Dose selection rationale: Based on study in 2004 giving LD50 of 891 mg/kg in males and 656 mg/kg in females
- Rationale for animal assignment (if not random):
- Other:

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Maternal bodyweights, first weight after GD=0 was GD = 6. After that every 3 days until GD = 21.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations:

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined: uterus, uterine contents, ovaries, fetuses,

OTHER:
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
- Soft tissue examinations: Yes
- Skeletal examinations: Yes:
- Head examinations: No data
Statistics:
Whenever possible, the data were presented as mean +/- SD. The number of corpora lutea, implantaion sites and live foetuses, and various body weights were analysed by one-way analysis of variance, followed by Dunnett's test if differences were found. The frequency of post-implantation loss, dead foetuses, resorptions and alterations among litterswas evaluated by using Kruskal-Wallis test followed by the Mann-Whitney test where appropriate. Rates of pregnancy and of litters with dead foetuses or resorptions and incidences of foetal alterations per dose were analysed by using Fishers's test. Where applicable, least-squares analysis was caried out. The reported level of statitsical significance was p < 0.05. The litter was used as the basis for the analysis of foetal variables.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Macroscopic changes in the liver. This mainly consisted of a pale and mottled organ.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
one dam from the 250 mg/kg dose group was found dead on GD 21. No obvious cause for death was established.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Reduced body weight gain.
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:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Foetal skeletal variations and delayed ossification in fore and hinf limbs and caudal vertebra centra.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed

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
Details on maternal toxic effects:
Macroscopic changes were noted in most of the dams at 150 mg/kg dose and higher.

Effect levels (maternal animals)

Key result
Dose descriptor:
dose level:
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
necropsy findings
Remarks on result:
other: Liver pale and mottled.

Maternal abnormalities

Key result
Abnormalities:
effects observed, treatment-related
Localisation:
other:
Description (incidence and severity):
Liver pale and mottled.

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
effects observed, treatment-related
Visceral malformations:
effects observed, treatment-related

Effect levels (fetuses)

Key result
Dose descriptor:
dose level:
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Reduced foetal body weight.
Remarks on result:
other: Dose related

Fetal abnormalities

Key result
Abnormalities:
effects observed, treatment-related
Localisation:
skeletal: skull
skeletal: forelimb
skeletal: sternum
skeletal: rib
skeletal: vertebra
skeletal: hindlimb
Description (incidence and severity):
Significantly delayed ossification at 250 mg/kg/day as evidenced by the reduced numbers of osified phalanged in fore and hind limbs, metatarsals in hindlimbs and caudal vertebral centra.

Overall developmental toxicity

Key result
Developmental effects observed:
yes
Lowest effective dose / conc.:
200 mg/kg bw/day (nominal)
Treatment related:
yes
Relation to maternal toxicity:
not specified
Dose response relationship:
yes
Relevant for humans:
yes

Applicant's summary and conclusion

Conclusions:
At 150 mg/kg/day, maternal response to treatment was indicated by macroscopic changes in the liver.