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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)

Data source

Reference
Reference Type:
publication
Title:
THE TERATOGENIC POTENTIAL IN RATS AND RABBITS OF D&C YELLOW NO. 8
Author:
C. M. BURNETI, E. I. GOLDENTHAL
Year:
1986
Bibliographic source:
Fd Chem. Toxic, Vol. 24, No. 8, pp. 819 823, 1986

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
Teratogenic toxicity study of D&C YELLOW NO. 8 in rats
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): D & C Yellow No. 8, sodium fluorescein (C.I. No. 45350)
- Molecular formula (if other than submission substance): C20H12O5.2Na
- Molecular weight (if other than submission substance): 376.274 g/mole
- Substance type: Organic
- Physical state: No data available
- Impurities (identity and concentrations): 13 %

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 18 weeks
- Weight at study initiation: 246 to 379 g
- Fasting period before study: No data available
- Housing: Animals were housed individually and identified by ear tag.
- Diet (e.g. ad libitum): Purina Certified Rodent Chow No. 5002, ad libitum
- Water (e.g. ad libitum): water, ad lib.
- Acclimation period: 4 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.22 ± 15
- Humidity (%):50 ± 15%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle.

IN-LIFE DATES: From: To: No data available

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: Doses were prepared daily in an aqueous solution at 100, 500 or 1500 mg/kg body weight.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Water
- Concentration in vehicle: 0, 100, 500 or 1500 mg/kg body weight.
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
14 days
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 500 or 1500 mg/kg body weight.
Basis:
nominal in water
No. of animals per sex per dose:
Total: 100
0 mg/kgbw/day:25 female
100 mg/kgbw/day:25 female
500 mg/kgbw/day:25 female
1500 mg/kgbw/day:25 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: No data available
- Rationale for animal assignment (if not random): Mated females were assigned to the control group or to one of the three treatment groups of 25 animals each.
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked in table [No.?] were included: Survival was observed.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: On days 6, 9, 12 and 16 of gestation

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

FOOD EFFICIENCY: No data available
- Body weight gain in kg/food consumption in kg per unit time X 100 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

OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: No data available
- Time schedule for collection of blood: No data available
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

CLINICAL CHEMISTRY: No data available
- Time schedule for collection of blood: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined. No data available

NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data available

OTHER: No data available
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
The thoracic and abdominal cavities and the internal organs of the dams were examined for gross morphological changes.

HISTOPATHOLOGY: No data available
Other examinations:
The number and location of viable and non-viable foetuses, early and late resorptions, total implantations and corpora lutea were recorded.
Statistics:
Statistical analysis were performed by using Chi-square test criterion with Yates" correction for 2 x 2 contingency lables and, or Fisher's exact probability test as described by Sicgel (1956) to Differences in the foetal sex distribution and the number of litters with malformations between control and treated groups. The numbers of early and late resorptions, dead foetuses and post-implantation losses were compared between groups by the Mann Whitney U test as described by Siegel (1956) and Well (1970). The mean numbers of viable foetuses, total implantations, corpora lutca and mean foetal weights were compared between groups by analysis of variance (one way classification), Bartlett's test for homogeneity of variances, and the appropriate t- test (for equal or unequal variances) as described by Steel & Torric (1960) using Dunnctt's multiple comparison tables (1964).

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
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
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
not specified
Details on results:
Mortality :
When treated with 1500 mg/kg bw/day, Six rats died during the dosing period as compared to control.

Clinical signs:
Orange discoloration of the urine was observed in all the treated rats as compared to control.

Body weight and weight gain When treated with 1500 mg/kg bw/day, decrease in body weight gain was observed in treated rats as compared to control.

Food consumption and compound intake No data available

Food efficiency No data available

Water consumption and compound intake: No data available

Opthalmoscopic examination No data available

Haematology: No data available

Clinical chemistry: No data available

Urinanalysis: No data available

Neurobehaviour: No data available

Organ weights No data available

Gross pathology: When treated with 1500 mg/kg bw/day, green discoloration of the amniotic fluid and green colored small intestines were observed in treated rats as compared to control.

When treated with 100 and 500 mg/kg bw/day, green discoloration of the amniotic fluid was observed in treated rats as compared to control.

Histopathology No data available

Details on results:
There were no biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations, number of foetuses or litters with developmental variations in any of the treated groups.

At 1500 mg/kg be/day, slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed as compared to control. However, these values fell within the ranges of historical control data.

Effect levels

Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: No effect on survival, clinical sign, body weight and body weight gain, gross pathology and reproductive performances

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Mean maternal body weights for rats given D & C Yellow No. 8 by gavage on days 6-19 of gestation

 

 

Mean maternal body weights (g)

Day of gestation

Dose (mg kg / day)

0 (control)

100

500

1500

0

 

246± 145

249 ± I3.7

249± 14.7

241± 12.4

6

 

272 ± 16.8

271 ± 14.4

276± 14.5 

263 ±12.2

9

 

279+ 18,1

278 ± 15.9

283 ± 16.7 

269 ± 16.5

12

 

294 ± 20.1

292 ± 17.5

296 ± 16.1

282 ± 16.9

16

 

321 ± 23.4

320 ± 18.9

323 ± 20.4

300 ± 30.7

20

 

379 ± 30.0

378 ± 21.2

374 + 32.2

359 ± 33.3

 

Mean maternal body-weight changes in rats given D & C Yellow No. 8 on days 6 -19 of gestation

 

 

Mean maternal body weights (g)

Day of gestation

Dose (mg kg / day)

0 (control)

100

500

1500

0 -6

 

26

22

27

22

6 -9

 

7

7

7

6

9 -12

 

15

14

13

13

12 -16

 

27

28

27

18

16 -20

 

58

58

51

59

6-20

 

107

107

98

06

0-20

 

133

129

125

118

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 500 mg/kg body weight /day when CD Sprague-Dawley female rats were treated with D&C YELLOW NO. 8.
Executive summary:

In a Teratogenic toxicity study, CD Sprague-Dawley female rats were treated with D&C YELLOW NO. 8 in the concentration of 0, 100, 500 and 1500 mg/kg body weight/ day by oral gavage.Six rats died during the dosing period at 1500 mg/kg bw/day and Orange discoloration of the urine was observed in all the treated rats as compared to control. Similarly, green discoloration of the amniotic fluid and green colored small intestines were observed at 1500 mg/kg bw/day and green discoloration of the amniotic fluid was observed at 100 and 500 mg/kg bw/day treated rats as compared to control. In addition, slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) were observed as compared to control. However, these values fell within the ranges of historical control data. There were no biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations,number of foetuses or litters with developmental variations in any of the treated groups. Therefore, NOAEL was considered to be 500 mg/kg body weight /day when CD Sprague-Dawley female rats were treated with D&C YELLOW NO. 8 orally by gavage from day 6 to 19 of gestation.