Calcium sulfate

Developmental toxicity / teratogenicity

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

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

no

Remarks:

Predates GLP

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Calcium sulfate

EC Number:

231-900-3

EC Name:

Calcium sulfate

Cas Number:

7778-18-9

Molecular formula:

CaSO4

IUPAC Name:

calcium sulfate

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 or test system 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 / concentrations

Doses / concentrations 1

Dose / conc.:

16 mg/kg bw/day

Remarks:

Species: mice, rats, rabbits

Doses / concentrations 2

Dose / conc.:

74.3 mg/kg bw/day

Remarks:

Species: mice, rats, rabbits

Doses / concentrations 3

Dose / conc.:

345 mg/kg bw/day

Remarks:

Species: mice, rats, rabbits

Doses / concentrations 4

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)

1

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.

2

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.

3

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	Pregnant^1
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	Pregnant^1
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	Pregnant^1
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.