Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
screening for reproductive / 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
Justification for type of information:
Data is from publication .

Data source

Reference
Reference Type:
publication
Title:
CHRONIC TOXICITY STUDIES ON FOOD COLOURS. PART II. OBSERVATIONS ON THE TOXICITY OF FD & C GREEN NO. 2 (LIGHT GREEN SF YELLOWISH), FD & C ORANGE NO. 2 (ORANGE SS) AND FD & C RED NO. 32 (OIL RED XO) IN RATS.
Author:
M. G. ALLMARK, H. C. GRICE AND W. A. MANNELL
Year:
1956
Bibliographic source:
J. Pharma. Pharmacology; Vol. 8, Pg. no. 417-424, 1956

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: As mention below
Principles of method if other than guideline:
Reproductive toxicity study was conducted to evaluate the reproductive effects of test substance Sudan II in rats for 20 weeks.
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Sudan II
- Molecular formula (if other than submission substance): C18H16N2O
- Molecular weight (if other than submission substance): 276.337 g/mol
- Substance type: Organic
- Physical state: Solid
- Purity: No data available
- 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): C.I. Solvent orange 7 (Sudan II) ,1- (2, 4-dimethylphenylazo)-2-naphthol; 1-[(2,4-dimethylphenyl)diazenyl]-2-naphthol
- Molecular formula: C18H16N2O
- Molecular weight: 276.337 g/mol
- Substance type: Organic
- Physical state: Solid

Test animals

Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: No data available
- Age at study initiation: 5 to 6 weeks
- Weight at study initiation: No data available - Fasting period before study: No data available
- Housing: Rats were kept in groups of two in a cage.
- Use of restrainers for preventing ingestion (if dermal): No data available
- Diet (e.g. ad libitum): Diet ad libitum
- Water (e.g. ad libitum): Water ad libitum
- 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

IN-LIFE DATES: No data available

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
not specified
Details on exposure:
Not specified.
Details on mating procedure:
Not specified.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
Not specified.
Duration of treatment / exposure:
20 weeks (More than 4 months)
Frequency of treatment:
Daily
Details on study schedule:
Not specified.
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
400 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10 animals/sex/dosein treated group
10 control animals were used for study.
Control animals:
yes, concurrent vehicle
Details on study design:
Not specified.
Positive control:
Not specified.

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data available
- Cage side observations checked in table [No.?] were included: Mortality was observed.


DETAILED CLINICAL OBSERVATIONS: No data available
- Time schedule: No data available

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight of the test animal was recorded weekly.

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


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


OTHER: Food efficiency of the test animal was also recorded weekly. For more accurate evaluation of food consumption it would have been preferable to place only one rat in a cage, but this was not possible.

Haematology determinations were made for 20 weeks on groups of male and female rats.
A slight modification of the pyridine-haemochromogen method of Rimington was used and the mean values of the final determinations were made. Blood haemoglobin parameter was examined.

Organs of surviving rats were weighed at the termination of the test. The mean weight in mg/g of body weight was calculated.

Estrous cyclicity (parental animals):
Not specified
Sperm parameters (parental animals):
Sperm parameters (parent animal)
Parameters examined in [all/P] male parental generations: A detailed examination was made of the haematoxylin-eosin stained paraffin sections of testes
Litter observations:
Not specified
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: No data available
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]: No data available
- Other: Post-mortem examinations were made where possible on rats which died on test. All surviving rats were killed at the end of the experiments and post-mortem examinations were made.

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]: No data available

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively: Yes, many of the organs were weighed and prepared for histological examination.
A detailed examination was made of the haematoxylin-eosin stained paraffin sections of a number of organs including lung, heart, liver, spleen, thyroid, pancreas, stomach, small intestine, kidney, urinary bladder, adrenal, testes, ovaries and thymus.
Postmortem examinations (offspring):
Not applicable
Statistics:
Standard deviation was observed.
Reproductive indices:
Not specified
Offspring viability indices:
Not specified

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
By the end of 20 weeks, 6 male rats receiving a dose of 200 mg/kg bw/day had died whereas all 10 female rats has survived for 20 weeks.

However, 5 male and 3 female rats receiving a dose of 400 mg/kg bw/day had died by the end of the experiment.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In a number of cases the body weights of the test animals were less than the controls. In other cases the body weights of test and control animals were about the same.


Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumption was affected by the test substance at the dose level of 400 mg/kg bw/day.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Food efficiencey was affected by the test substance at the dose level of 400 mg/kg bw/day.
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
A significant decline in blood haemoglobin values was found in all groups of both sexes at the dose level of both 200 and 400 mg/kg bw/day. This same result was also evident from an examination of the data obtained on each sex.
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:
effects observed, treatment-related
Description (incidence and severity):
When male rats was treated with 200mg/kgbw/day ,there were significant change in the mean organ weight of heart and adrenal.
When male rats was treated with 400mg/kgbw/day ,there were significant change in the mean organ weight of Lung heart ,liver, spleen, kidney, adrenal and testes.

When female rats was treated with 200mg/kgbw/day ,there were no significant change in the mean organ weight of Lung heart ,liver, spleen, kidney and adrenal.

When female rats were treated with 200mg/kgbw/day, there were significant change in the mean organ weight of liver and adrenal.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
No consistent histopathological changes were observed in the tissues or organs of the female test animal at dose level of 200and 400 mg/kgbw/day.

Altered spermatogenesis are observed in male test group receiving the dose 400 mg/kgbw/day compare 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:
effects observed, treatment-related
Description (incidence and severity):
No consistent histopathological changes were observed in the tissues or organs of the female test animal at dose level of 200and 400 mg/kgbw/day.

Altered spermatogenesis are observed in male test group receiving the dose 400 mg/kgbw/day compare to control.
Reproductive performance:
not specified

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: No significant effect were observed in histopathology and reproductive fuction

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
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:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P1)

Reproductive function: estrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
not specified

Results: F1 generation

Effect levels (F1)

Dose descriptor:
other: Not specified
Generation:
other: Not specified
Based on:
not specified
Sex:
not specified
Basis for effect level:
other: Not specified
Remarks on result:
other: Not specified

Target system / organ toxicity (F1)

Critical effects observed:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified

Any other information on results incl. tables

Observation on male rat

Product dosage

NO. of rats surviving/

NO. of rats on test

Sex

Mean body weight final

 

Mean hg

Mean organ weight mg/g

Histopathology

 

Testicles

Altered spermatogenesis

 

 

 

 

 

0 mg/kgbw/day

4/10

M

251.0±11.0

17.3±0.40

9.7±0.38

0

200 mg/kgbw/day

4/10

M

155.0±18.9*

14.1±0.81*

9.6±1.32

0

400mg/kgbw/day

5/10

M

160±14.0*

12.8±1.50*

9.6±1.65

3

Applicant's summary and conclusion

Conclusions:
When the test animal is exposed orally to the test substance Sudan II, No adverse effects were observed at a dose level of 200 mg/kg bw/day.
Executive summary:

Reproductive toxicity study was performed to evaluate the reproductive effects of the test substance on the test animals.

 

When the test animal is exposed orally (gavage)to the test substance Sudan II for 20 weeks at a dose conc. of 200 and400 mg/kg bw/day,No adverse effects were observed at a dose level of200 mg/kg bw/day.

 

Testicular changes are observed in animals receiving the higher concentrations i.e; 400 mg/kg bw/day.

 

A significant decline in blood haemoglobin values was found in all groups of both sexesat the dose level of both 200 and 400 mg/kg bw/day.

 

Food consumption and food efficiency was affected by the test substance at the dose level of 400 mg/kg bw/day.In a number of cases the organ weights were about the same as the controls but the body weights of the test animals were less than the controls, suggesting the possible utilization of muscle protein. In other cases the body weights of test and control animals were about the same but the organ weights differed significantly.

 

By the end of 20 weeks, 6 male rats receiving a dose of 200 mg/kg bw/day had died whereas all 10 female rats has survived for 20 weeks. However, 5 male and 3 female rats receiving a dose of 400 mg/kg bw/day had died by the end of the experiment.

 

Adetailed examination was made of the haematoxylin-eosin stained paraffin sections of a number of organs including lung, heart, liver, spleen, thyroid, pancreas, stomach, small intestine, kidney, urinary bladder, adrenal, testes, ovaries and thymus. Heart, liver, spleen, kidneys and testes were the organs chiefly affected.No consistent histopathological changes were observed in the tissues or organs of the test animal. Testicular changes are observed in animals receiving the higher concentrations. ThereforeSudan II was not considered to be a reprotox for male and female rats at a dose level of200 mg/kg bw/day.Hence it is not likely to be classiffied as reprotox.