Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
A study of the embyotoxixity of the food colour Ponceau 4R in rats
Author:
Meyer O. et al.
Year:
1975
Bibliographic source:
Toxicology 5, pp. 201-207

Materials and methods

Principles of method if other than guideline:
Embryotoxicity study in rats by gavage from day 1 to 20 of the gestation period. Foetuses were removed on day 21.
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Wistar
Remarks:
SPF
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 11 weeks
- Housing: steelwire cages


ENVIRONMENTAL CONDITIONS
- Temperature: 24 ± 1 °C
- Humidity: 60 ± 5 %
- Air changes: 8 per hr
- Photoperiod: electric light from 4 p.m. to 4 a.m.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: test substance dissolved in distilled water
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1:1
- Length of cohabitation: 5 days
- Proof of pregnancy: the presence of plug was checked every evening (day 0) and morning (day 1)
Duration of treatment / exposure:
From day 1 to 20.
Frequency of treatment:
Once per day.
Duration of test:
On day 21, rats were sacrificed.
Doses / concentrationsopen allclose all
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Dose / conc.:
2 000 mg/kg bw/day (actual dose received)
Dose / conc.:
4 000 mg/kg bw/day (actual dose received)
Control animals:
yes, concurrent no treatment
other: positive control (250 mg aspirin)
Details on study design:
- Dose selection rationale: based on range-finding test in which even the highest dose was tolerated without any deleterious effect.

Examinations

Maternal examinations:
BODY WEIGHT: Yes
- Time schedule for examinations: on day 21

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined: weight of intact uterus, number of corpora lutea, of implantations and of foetuses, alive or dead. Weight gain of the damns, sex ratio of foetuses and preimplantation loss were calculated.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: yes
- Number of corpora lutea (CL): yes
- Number of implantations: yes
- Preimplantation loss: yes
- Postimplantation loss: yes
- Number of foetuses, dead or alive, and sex ratio: yes
Fetal examinations:
- External examinations: yes, all per litter
- Soft tissue examinations: yes, half per litter
- Skeletal examinations: yes: half per litter
Statistics:
Student's t-test was performed on weight gain of the dams, mean weight of the intact uteri, preimplantation loss, and postimplantation loss. Critical values for testing two-by-two tables were performed on number of young with fourteenth rib(s) and Wilcoxon Ranks test on litter size.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Weight gain indicated a slight tendency to decrease with increasing doses.

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:
no effects observed

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
4 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
maternal abnormalities
number of abortions
pre and post implantation loss
total litter losses by resorption
effects on pregnancy duration
early or late resorptions
dead fetuses
changes in pregnancy duration
changes in number of pregnant
necropsy findings

Results (fetuses)

Fetal body weight changes:
not examined
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 litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
Skeletal and visceral malformations were sporadic and not dose-related. The incidence of retarded ossification and subcutaneous hemorrhage was distributed equally among the groups.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
4 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
changes in sex ratio
fetal/pup body weight changes
changes in litter size and weights
external malformations
skeletal malformations
visceral malformations

Overall developmental toxicity

Developmental effects observed:
no

Any other information on results incl. tables

Reproduction data

substance dose mg/kg bw/d pregnancy rate mean values per pregnant dam % loss dams with resorptions
corpora lutea implantations resorptions viable fetuses preimplantations ± s.d. post-implantations ± s.d. no.  %
test subs. 0 14/15 11.5 9.2 1.0 8.2 22.1 ± 27.0 27.1 ± 28.6 5 35.7
1000 13/15 12.3 10.3 0.7 9.6 17.2 ± 21.6 21.4 ± 20.4 6 46.2
2000 13/15 11.0 9.2 0.5 8.6 15.4 ± 22.9 27.1 ± 30.8 5 38.5
4000 12/15 11.5 9.3 1.3 8.1 20.6 ± 33.3 37.0 ± 38.4 7 58.3
aspirin 250 7/10 10.7 9.7 7.6 2.1 12.3 ± 35.7 81.0 ± 30.5 7 100

Litter data

substance

dose mg/kg

bw/d

viable young male/female mean pup weight g ± s.d.
total mean
test subs. 0 115 8.2 1.158 3.25 ± 1.08
1000 125 9.6 1.182 3.01 ± 0.61
2000 112 9.3 0.933 3.40 ± 0.86
4000 97 8.1 1.077 3.04 ± 0.67
aspirin 250 15 2.1 0.909 2.27 ± 0.58

Skeletal and internal malformations

substance dose mg/kg bw/d extra ribs foetuses with other skeletal malformations number examined findings (number)
litter foetus litters foetuses
test subs. 0 3/14 4/57 1 scoliosis, 1 kyphosis 13 58 hypoplasia of diaphragma (1), aplasia of left internal genital organ (1)
1000 6/13 10/65 0 13 60
2000 4/12 5/58 0 12 54 bilateral anophthalmia (1), unilateral anophthalmia (1), diaphragmatic hernia (1)
4000 5/12 5/52 0 10 45
aspirin 250 2/3 5/8 0 3 7

Applicant's summary and conclusion

Conclusions:
Under test condtions, no maternal toxicity or embryotoxicity was noted.
Executive summary:

Method

Embryotoxicity study in SPF Wistar rats, dosed by gavage from day 1 to 20 of gestation in doses of 0, 1000, 2000 and 4000 mg/kg bw/day, dissolved in distilled water. Foetus were removed on day 21 day. Aspirin at dose of 250 mg/kg bw/day was used as control.

Results

No effect was seen on: number of corpora lutea (CL), implantations, foetuses dead or alive, gross malformations, skeletal and internal malformations, weight of foetuses. The highest dose of 4000 mg/kg bw/d was identified as NOAEL.