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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
reproductive toxicity, other
Remarks:
Teratogenicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from publication

Data source

Reference
Reference Type:
publication
Title:
The teratogenic potential in rats and rabbits of test chemical
Author:
C. M. BURNETI.et.al
Year:
1986
Bibliographic source:
Food chemical toxicology,1986

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: as below
Principles of method if other than guideline:
Teratogenic Potential study of test material in rats by oral administration.
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
Details on test material:
Name of test material : D & C Yellow No. 8, sodium fluorescein
- Molecular formula : C20H10Na2O5
- Molecular weight : 376.274 g/mol
- Substance type: Organic
- Physical state: No data

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
CD
Details on species / strain selection:
No data available
Sex:
female
Details on test animals and environmental conditions:
Details on test animal
TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: (P) x wks; (F1) x wks: P- 18 weeks
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: P-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): Animals were housed individually
- Water (e.g. ad libitum): water, ad lib.
- Acclimation period: 4 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.22 ± 15 °C
- 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
Type of inhalation exposure (if applicable):
not specified
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with vehicle at dose levels of 0, 100, 500 or 1500 mg/kg body weight and prepared daily

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

VEHICLE
- Justification for use and choice of vehicle (if other than water):No data
- Concentration in vehicle:0, 100, 500 or 1500 mg/kgbw
- Amount of vehicle (if gavage):10 mL/Kg
- Lot/batch no. (if required):No data
- Purity:No data
Details on mating procedure:
- M/F ratio per cage: 1: 1.
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Copulatory plug or presence of sperm in vaginal washings was designated as day 0 of gestation
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
14 days (days 6-19 of gestation)
Frequency of treatment:
Daily
Details on study schedule:
not specified
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
500 mg/kg bw/day
Dose / conc.:
1 500 mg/kg bw/day
No. of animals per sex per dose:
Total: 100 females
0 mg/Kg bw: 25 females
100 mg/Kg bw: 25 females
500 mg/Kg bw: 25 females
1500 mg/Kg bw: 25 females
Control animals:
yes, concurrent vehicle
Details on study design:
not specified
Positive control:
not specified

Examinations

Parental animals: Observations and examinations:
Survival, clinical sign and body weights were examined.
Oestrous cyclicity (parental animals):
not specified
Sperm parameters (parental animals):
not specified
Litter observations:
sex and body weight were examined.
Postmortem examinations (parental animals):
Gross pathology were examined.
Postmortem examinations (offspring):
Gross external malformations and variations were examined.
Statistics:
Differences in the fetal sex distribution and the number of litters with malformations between control and treated groups wcre compared using the 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). 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 t\)etal weights were compared between groups by analysis of variance (oneway classification), Bartlett's test l\~r 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).
Reproductive indices:
Viable and non-viable foetuses, early and late resorptions, total implantations and corpora lutea were examined.
Offspring viability indices:
not specified

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Orange discoloration of the urine was observed in all the treated rats as compared to control.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
When treated with 1500 mg/kg bw, Six rats died during the dosing period as compared to control.
Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
When treated with 1500 mg/kg bw, slight reductions in body-weight gains as compared to controls, throughout the dosing period
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:
effects observed, treatment-related
Description (incidence and severity):
Orange discoloration of the urine was noted in all treated rats during the treatment period.

Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Green discoloration of the amniotic fluid was observed in 1, 10 and 16 rats at 100, 500 and 1500 mg/kg/day groups, respectively, and the small intestines were green in colour in many rats at 500 mg/Kg group.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No effect on Viable and non-viable foetuses, early and late resorptions, total implantations and corpora lutea and sex ratio of fetuses were observed as compared to control.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
1 500 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
mortality
body weight and weight gain
urinalysis
gross pathology
reproductive performance
other: No effect observed
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:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No effect on Viable and non-viable fetuses were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effect on Body weight of fetuses were observed as compared to control.
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
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
When treated with 1500 mg/kg bw, A slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed however, these values fell within the ranges of historical control data.

No biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations and number of fetuses or litters with developmental variations were observed in treated rats as compared to control.
Histopathological findings:
not specified
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:
1 500 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
body weight and weight gain
gross pathology
other: No effect 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

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

Any other information on results incl. tables

Summary of maternal and foetal observations in study of rats given test material by gavage on days 6-19 of gestation

 

  Parameter                  

 

 

                                        Observation

 

 

0(control)

           100

  500

1500

 

No.

 %

SD

No.

%

SD

No.

%

SD

No.

%

SD

 

Animals on study

 

 

25

 

-

 

-

 

25

 

84.0

 

-

 

25

-

 

25

25

-

-

Animals that were gravid

25

100.0

-

21

0.0

-

21

84.0

24

24

96.0

-

Animals that died gravid

0

0.0

-

0

100

-

0

0.0

6

6

24.0

-

Animals examined at caesarean section

25

100.0

-

25

16.0

-

25

100

19

19

76.0

-

Non-gravid

0

0.0

-

4

84.0

-

4

16

1

1

5.3

-

Gravid

25

100.0

-

21

-

2.55

21

84

18

18

94.7

-

Dams with viable foetuses

25

100.0

-

21

-

1.06

21

84

18

18

94.7

-

Viable foetuses/dam

13.3

-

2.92

13.1

-

2.16

12.6

-

13.9

13.9

-

1.30

Post-implantation loss/dam

0.9

-[

1.78

0.9

-

1.88

0.6

-

2.89

0.5

-

0.79

Total implantations/dam

14.2

-

2.24

14.0

-

-

13.1

-

2.13

14.4

-

1.10

Corpora lutea/dam

15.6

-

2.04

15.1

-

-

15.3

-

-

15.8

-

1.56

Foetal sex: male

                    Female

178

154

53.6

46.4

-

148

132

52.9

47.1

-

130

134

49.2

50.8

-

118

133

47.0

53.0

-

Mean foetal body weight (g)

3.4

-

0.29

3.5

-

0.36

3.5

-

0.36

3.5

-

0.45

Summary of the incidence of malformations and developmental and genetic variations

observed among foetuses from rats given test material by gavage on days 6-19 of gestation

Observations

Dose (mg/kg/day)…

No. of foetuses (no. of litters)

0 (control)

100

500

1500

No. of litters examined

No. of foetuses examined externally

No. of foetuses examined viscerally

No. of foetuses examined skeletally

 

25

332

99

21

275

81

21

264

93

18

251

70

Malformations observed

Skull anomaly

Thoracoschisis

Gastroschisis

Sternoschisis

Bent ribs

Radius bent

Carpal flexure

Foetal anasarca

Total foetuses (litters) with malformations:

 

 

2 (1)

1(1)

2 (1)

1 ( 1 )

1 (1)

2 (1)

 

2 (1)

3 (2)

 

 

 

 

 

 

 

6 (3)

 

 

 

6 (3)

 

 

 

 

 

 

 

1 (1)

 

1 (1)

 

 

 

 

 

 

 

 

 

0 (0)

Variations observed

27 presacral vertebrae 14th rudimentary rib(s)

14th lull rib(s)

12 full pairs of ribs with bilateral 13th

rudimentary ribs

 

 

 

3 (2)

27 (11)

2 (1)

 

 

1 (1)

 

 

23 (9)

 

1(1)

17 (8)

 

2 (1)

49 (14)

2 (1)

 

 

Skull reduced in ossification

Hyoid unossified

Vertebrae reduced in ossification

Femur reduced in ossification

Pubis unossified

lschium unossified

Femur unossified

Metatarsals unossified

Sternebrae no. 5 and/or no. 6 unossified

Other sternebrae unossified

Sternebrae misaligned

Major vessel variation

Renal papillae not developed and/or

distended ureter

 

 

4 (3)

7 (5)

2 (1)

1 (1)

2 (1)

2 (1)

1 (1)

1 (1)

 

33 (9)

 

 

 

 

 

8 (6)

 

4 (2)

4 (3)

 

 

 

 

 

 

 

18 (8)

 

 

 

 

 

2 (2)

 

5 (3)

2 (1)

 

 

 

 

 

 

 

33 (12)

1 (1)

1 (1)

1 (1)

 

 

1 (1)

 

4 (2)

9 (3)

 

 

 

 

 

 

 

29 (11)

9 (4)

1 (1)

 

 

 

2 (2)

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 1500 mg/kg/day for P and F1 generation when CD Sprague-Dawley female rats treated with test material orally by gavage for 14 days.
Executive summary:

In a Teratogenic Potential Test, CD Sprague-Dawley female rats treated with test material in the concentration of 0, 100, 500 or 1500 mg/kg bw orally by gavage in water for 14 days (days 6-19 of gestation). Six rats died during the dosing period at 1500 mg/kg bw as compared to control. Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye. Slight reductions in body-weight gains at 1500 mg/kg bw and Orange discoloration of the urine was noted in all treated rats during the treatment period. No effect on reproductive parameters such as Viable and non-viable fetuses, early and late resorptions, total implantations and corpora lutea and sex ratio of fetuses were observed in treated female rats as compared to control. Green discoloration of the amniotic fluid was observed in 1, 10 and 16 rats at 100, 500 and 1500 mg/kg/day groups, respectively, and the small intestines were green in colour in many rats at 500 mg/Kg group. In addition, No effect on Body weight of fetuses were observed as compared to control. A slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed at 1500 mg/kg bw however, these values fell within the ranges of historical control data. No biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations and number of fetuses or litters with developmental variations were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1500 mg/kg/day for P and F1 generation when CD Sprague-Dawley female rats treated with test material orally by gavage for 14 days.