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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study is well documented and comparabale to a guideline but not performed to GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1974
Report Date:
1974

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
GLP compliance:
no
Remarks:
Predates GLP
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Calcium sulfate
- Physical state: Solid (white powder)

Test animals

Species:
other: mice, rats and rabbits
Strain:
other: Albino CD-1 outbred mice; Wistar derived albino rats; Dutch-belted rabbits
Details on test animals and environmental conditions:
Mice:
Adult virgin mice were gang-housed in disposable plastic cages in temperature and humidty controlled quarters with free access to food and fresh tap water.

Rats:
Adult virgin rats were individually housed in mesh bottom cages in temperature and humidity-controlled quarters with free access to food and fresh tap water.

Rabbits:
Adult virgin rabbits were individually housed in mesh bottom cages in temperature and humidity-controlled quarters with free access to food and fresh tap water.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
not specified
Details on exposure:
Mice: beginning on Day 6 and continuing daily through Day 15 of gestation the females were dosed with the indicated doses.

Rats: beginning on Day 6 and continuing daily through Day 15 of gestation the females were dosed with the indicated doses.

Rabbits: beginning on Day 6 and continuing daily through Day 18 the females were dosed.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
None specified
Details on mating procedure:
Mice: mated with young adult males, and observation of the vaginal sperm plug was considered Day 0 of gestation.

Rats: mated with young adult males, and observation of the vaginal sperm plug was considered Day 0 of gestation.

Rabbits: On Day 0, each doe was given an injection of 0.4 mL of human chorionic gonadotrophin (400 IU) via the marginal ear vein. Three hours later, each doe was inseminated artificially with 0.3 mL of diluted semen from a proven donor buck using approximately 20E+06 motile sperm according to the procedure described by Vogin et al.
Duration of treatment / exposure:
Mice: 10 days
Rats: 10 days
Rabbits: 13 days
Frequency of treatment:
Daily
Duration of test:
Mice: 17 days
Rats: 20 days
Rabbits: 29 days
Doses / concentrationsopen allclose all
Dose / conc.:
16 mg/kg bw/day
Remarks:
Species: mice, rats, rabbits
Dose / conc.:
74.3 mg/kg bw/day
Remarks:
Species: mice, rats, rabbits
Dose / conc.:
345 mg/kg bw/day
Remarks:
Species: mice, rats, rabbits
Dose / conc.:
1 600 mg/kg bw/day
Remarks:
Species: mice, rats, rabbits
No. of animals per sex per dose:
Mice: Mated animals 24-30
Rats: Mated animals 25
Rabbits: Mated animals 20-22
Refer to tables 3 (mice) table 4 (rats) and table 5 (rabbits)
Control animals:
yes, sham-exposed
other: Asprin was used as the positive control in mice at a dose of 150 mg/kg and in rats at a dose of 250 mg/kg ... (see attached file)
Details on study design:
None specified

Examinations

Maternal examinations:
Mice:
Body weights were recorded on Days 0, 6, 11, 15 and 17 of gestation.
All animals were observed daily for appearance and behaviour with particular attention to food consumption and weight in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. The urogenital tract of each dam was examined in detail for anatomical abnormality.

Rats:
Body weights were recorded on Days 0, 6, 11, 15 and 20 of gestation.
All animals were observed daily for appearance and behaviour with particular attention to food consumption and weight in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. The urogenital tract of each dam was examined in detail for anatomical abnormality.

Rabbits:
Body weights were recorded on Days 0, 6, 12, 18 and 29 of gestation.
All animals were observed daily for appearance and behaviour with particular attention to food consumption and weight in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. The urogenital tract of each dam was examined in detail for anatomical abnormality.
Ovaries and uterine content:
Mice:
Number of implanation sites, resorption sites and live and dead foetuses were recorded.

Rats:
Number of implanation sites, resorption sites and live and dead foetuses were recorded.

Rabbits:
The number of corpora lutea, implanation sites, resorption sites and live and dead foetuses were recorded.
Fetal examinations:
Mice:
Body weights of live pups were recorded. All foetuses were examined for the presence of external congenital abnormalities. One third of the foetuses for each litter underwent detailed visceral examinations employing the Wilson technique. The remaining two-thirds were cleared in potassium hydroxide (KOH), stained with alizarin red S dye and examined for skeletal defects.

Rats:
Body weights of live pups were recorded. All foetuses were examined for the presence of external congenital abnormalities. One third of the foetuses for each litter underwent detailed visceral examinations employing the Wilson technique. The remaining two-thirds were cleared in potassium hydroxide (KOH), stained with alizarin red S dye and examined for skeletal defects.

Rabbits:
Body weights of live pups were recorded. All foetuses were examined for the presence of external congenital abnormalities. The live foetuses of each litter were then placed in an incubator for 24 h for the evaluation of neonatal survival. All surviving pups were sacrificed and all pups were examined for visceral abnormalities by dissection. All foetuses were then cleared in potassium hydroxide (KOH), stained with alizarin red S dye and examined for skeletal defects.
Statistics:
None specified
Indices:
None specified
Historical control data:
None specified

Results and discussion

Results: maternal animals

Maternal developmental toxicity

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

Details on maternal toxic effects:
Not applicable

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOAEL
Remarks:
Mice
Effect level:
1 600 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: no clearly discernable effects on nidation or on maternal or foetal survival.
Dose descriptor:
NOAEL
Remarks:
Rats
Effect level:
1 600 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: no clearly discernable effects on nidation or on maternal or foetal survival.
Dose descriptor:
NOAEL
Remarks:
Rabbits
Effect level:
1 600 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no clearly discernable effects on nidation or on maternal or foetal survival.

Maternal abnormalities

Abnormalities:
no effects observed

Results (fetuses)

Reduction in number of live offspring:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Not applicable

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
1 600 mg/kg bw/day
Based on:
test mat.
Sex:
not specified
Basis for effect level:
other: no clearly discernable effects on nidation or on maternal or foetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham-treated controls.

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Developmental effects observed:
no

Any other information on results incl. tables

Table 6: Fate summary for mice:

Group

Material

Dose**

Total

Surviving at term

Mated

Pregnant

Total

Pregnant1

341

Sham

0.0

25

22

25

22

342

Aspirin

150.0

25

19

25

19

343

FDA71 - 86

16.0

28

22

28

22

344

FDA71 – 86

74.3

24

22

24

22

345

FDA71 – 86

345.0

25

24

25

24

346

FDA71 - 86

1600.0

30

23

29

22

* positive control: 150 mg/kg

** administered as a water solution (10 mL per kg of body weight)

1) includes all dams examined at term

Table 7: Reproduction data for mice:

Group

341

342

343

344

345

346

Dose (mg/kg)

Sham

Aspirin**

16.0

74.3

345.0

1600.0

Pregnancies

Total No.

22

19

22

22

24

23

Died or aborted (before Day 17)

0

0

0

0

0

1

To term (on Day 17)

22

19

22

22

24

22

Live litters

Total No.*

21

19

22

21

24

21

Implant sites

Total No.

251

240

263

283

265

267

Average/dam*

11.4

12.6

12.0

12.9

11.0

12.1

Resorptions

Total No. *

19

8

5

12

21

14

Dams with 1 or more sites resorbed

6

5

4

2

9

7

Dams with all sites resorbed

1

-

-

1

-

1

Percent partial resorptions

27.3

26.3

18.2

9.09

37.5

31.8

Percent complete resorptions

4.55

-

-

4.55

-

4.55

Live foetuses

Total No.

229

224

255

268

243

250

Average/dam*

10.4

11.8

11.6

12.2

10.1

11.4

Sex ratio (M/F)

1.16

1.07

0.90

1.00

0.94

1.02

Dead foetuses

Total*

3

8

3

3

1

3

Dams with 1 or more dead

2

6

2

3

1

3

Dams with all dead

-

-

-

-

-

-

Percent partial dead

9.09

31.6

9.09

13.6

4.17

13.6

Percent all dead

-

-

-

-

-

-

Average Foetus weight, g.

0.89

0.87

0.86

0.87

0.91

0.88

* includes only those dams examined at term

** positive control, 150.0 mg/kg

Table 8: Fate summary for rats:

Group

Material

Dose**

Total

Surviving at term

Mated

Pregnant

Total

Pregnant1

341

Sham

0.0

25

25

25

25

342

Aspirin

250.0

25

23

25

23

343

FDA71 - 86

16.0

25

21

25

21

344

FDA71 – 86

74.3

25

23

25

23

345

FDA71 – 86

345.0

25

23

25

23

346

FDA71 - 86

1600.0

25

21

25

21

* positive control 250 mg/kg

** administered as a water solution

1) includes all dams examined at term

Table 9: Reproduction data for rats.

Group

341

342

343

344

345

346

Dose

Sham

Aspirin**

16.0

74.3

345.0

1600.0

Pregnancies

Total No.

25

23

21

23

23

21

Died or aborted (before Day 20)

0

0

0

0

0

0

To term (on Day 20)

25

23

21

23

23

21

Live litters

Total No.*

25

18

21

23

23

21

Implant sites

Total No.

279

231

229

270

253

233

Average/dam*

11.2

10.0

10.9

11.7

11.0

11.1

Resorptions

Total No. *

4

62

4

7

7

4

Dams with 1 or more sites resorbed

3

11

3

3

2

3

Dams with all sites resorbed

-

4

-

-

-

-

Percent partial resorptions

12.0

47.8

14.3

13.0

8.70

14.3

Percent complete resorptions

-

17.4

-

-

-

-

Live foetuses

Total No.

275

168

224

263

246

229

Average/dam*

11.0

7.30

10.7

11.4

10.7

10.9

Sex ratio (M/F)

1.07

1.06

0.87

1.07

1.18

0.75

Dead foetuses

Total*

-

1

1

-

-

-

Dams with 1 or more dead

-

1

1

-

-

-

Dams with all dead

-

1

-

-

-

-

Percent partial dead

-

4.35

4.76

-

-

-

Percent all dead

-

4.35

-

-

-

-

Average Foetus weight, g.

3.68

2.39

3.92

3.94

3.77

4.04

* includes only those dams examined at term

** positive control: 250 mg/kg

Table 10: Fate summary for rats

Group

Material

Dose**

mg/kg

Total

Surviving at term

Mated

Pregnant

Total

Pregnant1

341

Sham

0.0

18

13

18

13

342

6-AN*

2.5

20

10

19

10

343

FDA71 - 86

16.0

22

14

19

11

344

FDA71 - 86

74.3

22

13

21

13

345

FDA71 - 86

345.0

20

13

19

12

346

FDA71 - 86

1600.0

20

14

15

11

* positive contorl: 2.5 mg/kg of 6 aminonicotinamide dosed on Day 9

** administered as a water solution.

1) includes all dams examined at term.

Table 11: Reproduction data for rabbits

Group

341

342

343

344

345

346

Dose

Sham

6-AN**

16.0

74.3

345.0

1600.0

Pregnancies

Total No.

13

10

14

13

13

14

Died or aborted (before Day 29)

0

1

3

0

1

3

To term (on Day 29)

13

10

11

13

12

11

Corpora lutea

Total No.

174

122

195

173

134

139

Average/dam mated

9.67

6.42

9.29

8.65

7.05

7.72

Live litters

Total No.

12

10

11

13

10

10

Implant sites

Total No.

69

56

60

70

68

62

Average/dam

5.31

5.60

5.45

5.38

5.67

5.64

Resorptions

Total No.*

4

2

4

7

9

4

Dams with 1 or more sites resorbed

3

1

3

3

6

1

Dams with all sites resorbed

1

-

-

-

2

1

Percent partial resorptions

23.1

10.0

27.3

23.1

50.0

9.09

Percent complete resorptions

7.69

-

-

-

16.7

9.09

Live foetuses

Total No.*

65

54

56

63

59

58

Average/dam

5.00

5.40

5.09

4.85

4.92

5.27

Sex ratio (M/F)

0.86

0.74

1.33

1.30

1.36

1.42

Dead foetuses

Total No.*

-

-

-

-

-

-

Dams with 1 or more dead

-

-

-

-

-

-

Dams with all dead

-

-

-

-

-

-

Percent partial dead

-

-

-

-

-

-

Percent all dead

-

-

-

-

-

-

Average foetus weight

38.2

33.8

37.8

38.2

39.9

42.4

* included only those dams examined at term

** positive control: 2.5 mg/kg of 6-aminonicotinamide dosed on Day 9

Applicant's summary and conclusion

Conclusions:
Mice:
Administration of up to 1600 mg/kg (body weight) of the test material to pregnant mice for 10 consecutive days had no clearly discernable effects on nidation or on maternal or foetal survival.

Rats:
The administration of up to 1600 mg/kg (body weight) of the test material to pregnant rats for 10 consecutive days had no clearly discernable effects on nidation or on maternal or foetal survival.

Rabbits:
up to 1600 mg/kg (body weight) of the test material to pregnant rabbits for 13 consecutive days had no clearly discernable effects on nidation or on maternal or foetal survival.

All species: The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham-treated controls.