Registration Dossier

Toxicological information

Developmental toxicity / teratogenicity

Currently viewing:

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:
other: GLP-study according to OECD test guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
minor deviations (typos, delay of draft report, ...) with no effect on the results and integrity of the study
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report): C.I. Pigment Red 254 (Cinilex® DPP Red SR1C)
- Substance type: pigment
- Physical state: powder (red)
- Analytical purity: > 99%, typically 99.4%
- Storage condition of test material: Room temperature

Test animals

Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: OXI-COOP ZRT. 1103 Budapest, Cserkesz u. 90.
- Age at study initiation: females 8.5-10 weeks, males at least 12 weeks
- Weight at study initiation: females 153 -218g
- Fasting period before study: no
- Housing: pre-mating period: 2-3 females /cage, 2-3 males/cage
during mating hours: 1 male with 1- 3 females
during pregnancy: 2-3 sperm positive females /cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 8 days for females, at least 27 days for males

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
0.5% Aqueous methylcellulose
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): standard vehicle
- Amount of vehicle (if gavage): treatment volume was 10 ml/kg bw
- Lot/batch no. (if required):N83746634
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The formulation was analyzed for its test item content by the Analytical Laboratory of
Toxi-Coop Zrt. Analysis of the formulation was performed twice during the study, on the first and last week of treatment.
Results were expressed in terms of percentage of the nominal concentration. The deviation from the nominal concentration was within the range of -9% and + 6%. A report of the analytical procedures and results in tabulated form are included in the study report.
The registered substance was stable in 0.5% Methylcellulose formulations at room temperature for up to 24 hours and at 2-8 ºC for up to three days
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1 male with 1-3 females
- Length of cohabitation: 2-4 hours/d until mating
- Further matings after two unsuccessful attempts: no
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
15 days (from gestational day 5 to 19)
Frequency of treatment:
daily
Duration of test:
20 days (after prood of pregancy)
Doses / concentrations
Remarks:
Doses / Concentrations:
250, 500, 1000 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
at least 20 females per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected with agreement of the Sponsor based on literature data presented by the Sponsor (MSDS).

Examinations

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

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: sperm positive females: on gestation days 0, 3, 5, 8, 11, 14, 17 and 20

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: Gross pathology observation
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 /
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter
Statistics:
Means and standard deviations and/or percentages were calculated.
Statistical analysis was performed with SPSS PC+ software.
The heterogeneity of variance between groups was checked by Bartlett's homogeneity of variance test. Where no significant heterogeneity is detected, a one-way analysis of variance was carried out. If the obtained result is positive, Duncan's Multiple Range test was used to assess the significance of inter-group differences.
Where significant heterogeneity is found, the normal distribution of data was examined by Kolmogorov-Smirnov test. In case of a none-normal distribution, the non-parametric method of Kruskal-Wallis One-Way analysis of variance was used. If there is a positive result, the inter-group comparisons are performed using the Mann-Whitney U-test.
Chi2 test was performed if feasible (8).
Indices:
Pre-implantation loss
Post-implantation mortality
Sex distribution
External abnormalities/litter
Visceral abnormalities/litter
Skeletal abnormalities/litter

Results and discussion

Results: maternal animals

Maternal developmental toxicity

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

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects

Results (fetuses)

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

Details on embryotoxic / teratogenic effects:
External examination:
One fetus was supposed to have thinned eyelids in the 250 mg/kg bw/day dose group however this was not proven by visceral examination. There were no malformed fetuses found in the experimental groups during the external examination.

Visceral examination:
Visceral malformations such as dilated lateral brain ventricles were observed in one fetus in the 500 mg/kg bw/day groups. An enlarged left ventricle of heart was recorded for one fetus in the 250 mg/kg bw/day groups. Considering the low incidence and lack of dose response, these alterations were not attributed to an effect of the test item.

Skeletal examination:
Malformations such as misshapen sternebra in one fetus in the 250 mg/kg bw/day group and a hemicentric thoracic vertebral centrum in the 1000 mg/kg bw/day dose groups were recorded at the skeletal examinations. Concidering the low incidence these abnormalities were not attributed to the administration of the test item

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
In a GLP-study according to OECD test guideline 414, the registered substance, orally applied, did not reveal any adverse effects on the pregnancy, body weight, food consumption data of the dams and on the intrauterine development of the fetuses. Therefore, both the NOAEL maternal toxicity and the NOAEL developmental toxicity were 1000mg/kg bw/day.
Executive summary:

The developmental toxicity of registered substance was investigated in a GLP-study according to OECD test guideline 414. Oral treatment of pregnant Hsd. Brl. Han: WISTAR rats from gestation day 5 up to the day before Caesarean section with C.I. Pigment Red 254 at the dose levels of 250, 500 and 1000 mg/kg bw/day did not reveal any adverse effects on the pregnancy, body weight, food consumption data of the dams and on the intrauterine development of the fetuses.
Under the conditions applied in this study and from the observations made in the dams and their fetuses the following no-observable-adverse-effect levels were derived:


NOAELmaternal toxicity:1000mg/kg bw/day

NOAELdevelopmental toxicity:1000mg/kg bw/day