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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
multi-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data from secondary source

Data source

Reference
Reference Type:
secondary source
Title:
A Multigeneration Study in Rats of D&C Red 33 in the Diet
Author:
European Commission
Year:
2007
Bibliographic source:
Scientific Committee on Consumer Products (SCCP), OPINION ON Acid Red 33, COLIPA n° C22, 2007 page no -19

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Multigeneration Study of D&C Red 33 in rats
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available

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): D&C Red 33
- Molecular formula : C16H11N3Na2O7S2
- Molecular weight: 467 g/mole
- Substance type: Organic
- Physical state: Powder
- Impurities (identity and concentrations): No data available
Specific details on test material used for the study:
- Name of test material (as cited in study report): D&C Red 33
- Molecular formula : C16H11N3Na2O7S2
- Molecular weight: 467 g/mole
- Substance type: Organic
- Physical state: Powder
- Impurities (identity and concentrations): No data available

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals and environmental conditions:
- Source: No data available
- Age at study initiation: (P) x wks; (F1) x wks: No data available
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: No data available
- Fasting period before study: No data available
- Housing: No data available
- Diet (e.g. ad libitum): Purina Rodent Chow 5002, ad libitum
- Water (e.g. ad libitum): No data available
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

Administration / exposure

Route of administration:
oral: feed
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: Purina Rodent Chow 5002
Details on mating procedure:
- M/F ratio per cage:: No data available

- Length of cohabitation: No data available

- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy No data available

- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.: No data available

- Further matings after two unsuccessful attempts: [no / yes (explain)]: No data available

- After successful mating each pregnant female was caged (how):: No data available

- Any other deviations from standard protocol:: F0 parental animals were mated twice (each group 20 males and 20 females), to produce two litters and the F1 parents were mated to produce three litters and the F2 parents were mated once to produce the F3a litters.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
110 days and more
Frequency of treatment:
Daily
Details on study schedule:
Details on study schedule
F0: 20 animals per sex per dose level
F1a: mating on day 100
F1b and F1c: mating at least 10 day later
F2a: mating of 20 F1 animals randomly selected
F3: mating of 20 F2b litters
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 0.25, 2.5, 7.5 and 25.0 mg/kg bw/day
Basis:

No. of animals per sex per dose:
Total: 600
F0 generation
0 mg/kg bw/day: 20 male, 20 female
0.25 mg/kg bw/day: 20 male, 20 female
2.5 mg/kg bw/day: 20 male, 20 female
7.5 mg/kg bw/day: 20 male, 20 female
25.0 mg/kg bw/day: 20 male, 20 female

F1 generation
0 mg/kg bw/day: 20 male, 20 female
0.25 mg/kg bw/day: 20 male, 20 female
2.5 mg/kg bw/day: 20 male, 20 female
7.5 mg/kg bw/day: 20 male, 20 female
25.0 mg/kg bw/day: 20 male, 20 female

F2 generation
0 mg/kg bw/day: 20 male, 20 female
0.25 mg/kg bw/day: 20 male, 20 female
2.5 mg/kg bw/day: 20 male, 20 female
7.5 mg/kg bw/day: 20 male, 20 female
25.0 mg/kg bw/day: 20 male, 20 female
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes

- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations:

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

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

OTHER:
Estrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
No data available
Postmortem examinations (parental animals):
Histopathology were examined.
Postmortem examinations (offspring):
Histopathology were examined.
Statistics:
No data available
Reproductive indices:
Fertility indices, gestation anomalies, viability and survival of the pups were examined.
Offspring viability indices:
No data available

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
When treated with 25 mg/kg bw/day, discoloration (pink or reddish) of the urine were observed in treated male and female rats as compared to control.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No effect on survival of treated rats were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effect on body weight of treated rats was observed as compared to control.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effect on food consumption of treated rats was observed as compared to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No histopathological changes were observed intreated rats as compared to control.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: estrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No effect on fertility indices, gestation anomalies, viability and survival of the pups were observed in treated rats as compared to control.

Details on results (P0)

Mortality: No effect on survival of treated rats were observed as compared to control.

Clinical signs: When treated with 25 mg/kg bw/day, discoloration (pink or reddish) of the urine were observed in treated male and female rats as compared to control.

Body weight: No effect on body weight of treated rats was observed as compared to control.

Food consumption:No effect on food consumption of treated rats was observed as compared to control.

Test substance intake: No data available

Reproductive function: estrous cycle: No data available

Reproductive function: sperm measures: No data available

Reproductive performance: No effect on fertility indices, gestation anomalies, viability and survival of the pups were observed in treated rats as compared to control.

Organ weights No data available

Gross pathology: No data available

Histopathology: No histopathological changes were observed intreated rats as compared to control.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on survival, clinical sign, Body weight, food consumption, histopathology and reproductive parameters
Remarks on result:
other: No effects on reproductive performance

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Description (incidence and severity):
When treated with 25 mg/kg bw/day, discoloration (pink or reddish) of the urine were observed in F1treated male and female pups as compared to control.
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No effect on survival of treated F1 pups were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effect on body weight of F1 treated male and female pups were observed as compared to control.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effect on food consumption of treated F1 rats were observed as compared to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
no effects observed
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on survival, clinical sign, Body weight, food consumption, histopathology and reproductive parameters
Remarks on result:
other: No toxic effects were observed

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F2 generation

General toxicity (F2)

Clinical signs:
no effects observed
Description (incidence and severity):
When treated with 25 mg/kg bw/day, discoloration (pink or reddish) of the urine were observed in F2 treated male and female pups as compared to control.
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No effect on survival of treated F2 pups were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effect on body weight of F2 treated male and female pups were observed as compared to control.
Food consumption and compound intake (if feeding study):
not specified
Description (incidence and severity):
No effect on food consumption of treated F2 rats were observed as compared to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Description (incidence and severity):
In F2a generation, 1 pup had exencephaly, spina bifida and great vessel anomalies which was considered an incidental finding.
Other effects:
no effects observed
Description (incidence and severity):
No effect on viability and survival of the pups were observed in treated rats as compared to control.

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not specified

Effect levels (F2)

Dose descriptor:
NOAEL
Generation:
F2
Effect level:
25 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on survival, clinical sign, Body weight, food consumption, histopathology and reproductive parameters
Remarks on result:
other: No toxic effects were observed

Target system / organ toxicity (F2)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 25 mg/kg bw/day when Charles River COBS CD (Sprague Dawley) male and female rats were treated wtih D&C Red 33.
Executive summary:

In a Multigeneration Study, Charles River COBS CD (Sprague Dawley) male and female rats were treated wtih D&C Red 33 in the concentration of 0, 0.25, 2.5, 7.5 and 25.0 mg/kg bw/day orally in diet. One control groups and four test groups (100 males and 100 females) were used. The control and treated rats were maintained on their respective diets for the duration of this generation. F0 parental animals were mated twice (each group 20 males and 20 females), to produce two litters and the F1 parents were mated to produce three litters and the F2 parents were mated once to produce the F3a litters. All the animals were observed for toxicity signs, changes in behaviour and mortality. Body weight and parental food consumption were noted. Reproductive parameters were evaluated to determine male and female fertility indices, gestation anomalies, viability and survival of the pups. Histopathology (14 representative tissues) was performed from control and high dose group from F1 and F3a generation

 No effect on survival of F0, F1 and F2 generation were observed. Discoloration (pink or reddish) of the urine were observed in F0, F1 and F2 treated male and female rats as compared to control. Similarly, no effect on body weight and food consumption of treated F0, F1 and F2 rats were observed as compared to control. No effect on fertility indices, gestation anomalies, viability and survival of the pups were observed in F0, F1 and F2treated rats as compared to control. In addition no effect on histopathology of F0 and F1 treated rats were observed. In F2a generation, 1 pup had exencephaly, spina bifida and great vessel anomalies which were considered an incidental finding. Therefore, NOAEL was considered to be 25 mg/kg bw/day when Charles River COBS CD (Sprague Dawley) male and female rats were treated with D&C Red 33.