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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
Study period:
1971/1972
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented study report which meets basic scientific principles

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1972
Report Date:
1972
Reference Type:
publication
Title:
Toxicology of Indigo - a review
Author:
Ferber KH
Year:
1987
Bibliographic source:
J. Environ. Pathol. Toxicol. Oncol. 7, 73 -84 (1987)
Reference Type:
publication
Title:
Teratogenic Studies with Certified Colors in Rats and Rabbits.
Author:
Burnett CM et al.
Year:
1974
Bibliographic source:
Toxicol  Appl Pharmacol. 1974;29:121-122

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): D&C Blue No. 6
- Substance type: active compound
- Physical state: solid
- Analytical purity: 96%
- Lot/batch No.: Y-2206 and Y-2678
- FDA certification no.: Y-PMA-1

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (obtained as weanlings)
- Age at study initiation: satisfactory age for mating (about 8 weeks)
- Weight at study initiation: -
- Fasting period before study: -
- Housing: single
- Diet: Purina Laboratory Chow ad libitum
- Water: tap ad libitum
- Acclimation period: ca. 5 weeks

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: methylcellulose 0.5%
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
suspension in 0.5% methylcellulose


VEHICLE
- Dow Methocel 90 HG, 15000 CPS
- Justification for use and choice of vehicle (if other than water): TS is insoluble in water
- Concentration in vehicle: 50 mg/mL
- Amount of vehicle: 10 mL/kg (control and high dose), 3.2 mL/kg, 1 mL/kg
- Lot/batch no.: MM 101012K

POSITIVE CONTROL
- Acetylsalicylic acid (Aspirin)
- Supplier: Mallinckrodt
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- 1 M/1 F per cage
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
Duration of treatment / exposure:
Day 6 through 15 of gestation
Frequency of treatment:
daily
Duration of test:
21 days
Doses / concentrations
Remarks:
Doses / Concentrations:
50, 160, 500 mg/kg/day
Basis:
other: nominal in vehicle
No. of animals per sex per dose:
20 pregnant females
Control animals:
yes, concurrent vehicle
other: positive control: aspirin
Details on study design:
3 dosage levels were run concurrently with 2 negative (vehicle) and one positive (Aspirin) control groups

Examinations

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

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: -

BODY WEIGHT: Yes
- Time schedule for examinations: Days 0, 6, 15, 20 of gestation

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of resorptions: Yes
- Number viable and dead fetuses: Yes
- Distribution by sex: Yes
- Mean litter size: Yes
Fetal examinations:
- Mean fetal weight
- Determination of sex
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: one-third per litter
- Skeletal examinations: Yes: two-third per litter
Statistics:
Frequency of occurrence and seriousness of anomaly compared to control groups
Indices:
no data
Historical control data:
no data

Results and discussion

Results: maternal animals

Maternal developmental toxicity

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

Details on maternal toxic effects:
-

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOEL
Effect level:
500 mg/kg bw/day
Based on:
act. ingr.
Basis for effect level:
other: developmental toxicity
Dose descriptor:
NOEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
act. ingr.
Basis for effect level:
other: maternal toxicity

Results (fetuses)

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

Details on embryotoxic / teratogenic effects:
-

Effect levels (fetuses)

Dose descriptor:
NOEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
act. ingr.
Basis for effect level:
other: teratogenicity

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
D&C Blue No. 6 administered by gavage to groups of 20 female rats each during days 6 through 15 of gestation at dose levels of 50, 160, and 500 mg/kg/day was without effect on reproductive performance, maternal weight gain, and fetal development.
Executive summary:

Groups of 20 pregnant female Charles River CD rats received suspensions of D&C Blue No. 6 by stomach tube at doses of 500, 160 and 50 mg/kg/day from day 6 through day 15 of gestation. Comparable groups received either the methylcellulose suspending vehicle as negative controls or aspirin, 250 mg/kg/day as positive controls. On day 20 of gestation Caesarean section was performed, the uterus examined and fetuses processed for subsequent examination for skeletal or visceral anomalies.

On the basis of numbers of viable and dead fetuses, resorption sites, mean fetal weight, distribution by sex, mean litter size, frequency of skeletal and visceral or structural anomalies; weight gain of pregnant females: Indigo was without effect on reproductive performance, maternal weight gain and fetal development.