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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
Study period:
19/7/74 - 23/08/74
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Meets generally accepted scientific standards with acceptable restrictions. Noted deficiencies include; Food consumption not reported, Uterine weights not determined, One third used for visceral examination; should be 50%, Test substance identification (Batch etc) missing, No details on housing conditions/source of animals, administration of the test material was only during periods of organogenesis and not until day before pregnancy
Cross-reference
Reason / purpose:
reference to same study
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23/7/74 - 27/8/74
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Meets generally accepted scientific standards with acceptable restrictions. Noted deficiencies include; Food consumption not reported, Uterine weights not determined, One third used for visceral examination; should be 50%, Test substance identification (Batch etc) missing, No details on housing conditions/source of animals, administration of the test material was only during periods of organogenesis and not until day before pregnancy
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
other: no data
Deviations:
not specified
Principles of method if other than guideline:
Adult female albino (Wistar derived stock) rats were mated with young adult males. Observation of a vaginal sperm plug was considered as Day 0 of gestation. Dosing by oral intubation with a control (Vehicle at level equivalent to group receiving the highest dose or aspirin at 250 mg/kg) or test article in a water suspension at 2.4, 11.1, 51.7 and 240.0 mg/kg was carried out daily on Days 6 to 15 of gestation. Observations of body weight, appearence, behaviour, and food consumption were performed. Daily room temperature was recorded. On Day 20 of gestation all dams underwent Caesarean section. Sex, number of corpora lutea, implantation sites, resorption sites and live/dead foetuses recorded. Body weights of live pups recorded. Urogenital tract of each dam examined for anatomical normality. All foetuses examined grossly for presence of external congenital abnormalities. One third foetuses of each litter underwent detailed visceral examination and the remaining two thirds were cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.
GLP compliance:
no
Remarks:
Study predates GLP
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Outbred
- Age at study initiation: No data
- Weight at study initiation: 223 - 231 g
- Fasting period before study: No data
- Housing: Individual housing in mesh bottom cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: No data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 - 26.7
- Humidity (%): 64 - 78

IN-LIFE DATES: 23/7/74 - 27/8/74
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
VEHICLE: Water
- Amount of vehicle (if gavage): 1 mL/kg bw at doses equal to or below 250 mg/kg bw and 2 mL/kg at doses up to 500 mg/kg bw
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: No data
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
Duration of treatment / exposure:
10 days (Day 6 to Day 15 of gestation)
Frequency of treatment:
Daily
Duration of test:
20 days
No. of animals per sex per dose:
Table 1 Number of animals dosed

Material Dose (mg/kg) Total
Mated Pregnant
Sham 0.0 25 20
Aspirin 250.0 21 21
FDA 73-3 2.4 25 20
11.1 22 20
51.7 20 20
240.0 22 20

Control animals:
yes, sham-exposed
other: positive control: 250 mg/kg aspirin
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Appearence, behaviour, food consumption and weight observed daily.
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
- Time schedule for examinations: Body weights recorded on days 0, 6, 11, 15 and 20.
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus and urogenital tract
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes
Fetal examinations:
- External examinations: Yes: one third per litter
- Soft tissue examinations: Yes: one third per litter
- Skeletal examinations: Yes: two thirds per litter
- Head examinations: Yes: two thirds per litter
Statistics:
No data
Indices:
No data
Historical control data:
No
Details on maternal toxic effects:
Maternal toxic effects:no effects
Key result
Dose descriptor:
NOAEL
Effect level:
> 240 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no effects observed
Key result
Abnormalities:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Dose descriptor:
NOAEL
Effect level:
> 240 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Abnormalities:
no effects observed
Developmental effects observed:
no

Table 2 - Reproduction data

Dose (mg/kg)

Sham

Aspirin

2.4

11.1

51.7

240.0

Pregnancies

 

 

 

 

 

 

Total No.

20

21

20

20

20

20

Died or aborted (before Day 20)

0

1

0

0

0

0

To term (on Day 20)

20

20

20

20

20

20

Corpora Lutea

 

 

 

 

 

 

Total no.

255

267

247

258

243

245

Average/dam mated

10.2

13.4

9.88

11.7

12.2

11.1

Live litters

 

 

 

 

 

 

Total No.*

20

18

20

20

20

20

Implant Sites

 

 

 

 

 

 

Total No.

243

234

226

229

214

228

Average/dam*

12.2

11.7

11.3

11.5

10.7

11.4

Resorptions

 

 

 

 

 

 

Total No*

2

75

3

13

6

1

Dams with 1 or more sites resorbed

2

13

3

5

3

1

Dams with all sites resorbed

--

2

--

--

--

--

Per cent partial resorptions

10.0

65.0

15.0

25.0

15.0

5.00

Per cent complete resorptions

--

10.0

--

--

--

--

Live foetuses

 

 

 

 

 

 

Total No

241

159

223

216

208

227

Average/dam*

12.1

7.95

11.2

10.8

10.4

11.4

Sex ratio (M/F)

1.04

1.04

1.23

1.43

1.29

0.99

Dead Foetuses

 

 

 

 

 

 

Total No.*

--

--

--

--

--

--

Dams with 1 or more dead

--

--

--

--

--

--

Dams with all dead

--

--

--

--

--

--

Per cent partial dead

--

--

--

--

--

--

Per cent all dead

--

--

--

--

--

--

Average foetus weight (g)

3.76

2.45

4.07

3.91

3.96

3.89

* Includes only those dams examined at term

** Positive control: 250 mg/kg

 

Table 3 - Summary of skeletal findings

Findings

Dose (mg/kg)

Sham

Aspirin

2.4

11.1

51.7

240.0

Live foetuses examined (at term)

168/20

112/18

160/20

149/20

142/20

158/20

Sternebrae

 

 

 

 

 

 

Incomplete oss.

70/19

84/15

45/16

44/15

43/15

50/17

Scrambled

 

 

 

 

 

 

Bipartite

 

10/7

 

1/1

 

1/1

Fused

 

 

 

 

 

 

Extra

 

 

 

 

 

 

Missing

26/8

107/18

14/6

16/6

10/6

11/7

Other

 

 

 

 

 

 

Ribs

 

 

 

 

 

 

Incomplete oss.

 

 

 

 

 

 

Fused/split

 

4/3

 

 

 

 

Wavy

17/8

49/14

15/10

38/12

23/9

26/10

Less than 12

 

 

 

 

 

 

More than 13

1/1

102/17

1/1

1/1

 

 

Other

 

 

 

 

 

 

Vertebrae

 

 

 

 

 

 

Incomplete oss.

12/6

80/17

7/5

12/5

3/3

7/3

Scrambled

 

 

 

 

 

 

Fused

 

 

 

 

 

 

Extra ctrs. oss.

 

 

 

 

 

 

Scoliosis

 

 

 

 

 

 

Tail defects

 

 

 

 

 

 

Other

 

 

 

 

 

 

Skull

 

 

 

 

 

 

Incomplete closure

34/13

53/15

22/11

34/11

20/9

24/8

Missing

 

4/2

 

 

 

 

Craniostosis

 

 

 

 

 

 

Other

 

 

 

 

 

 

Extremities

 

 

 

 

 

 

Incomplete oss.

 

14/7

 

 

 

 

Missing

 

 

 

 

 

 

Extra

 

 

 

 

 

 

Miscellaneous

 

 

 

 

 

 

Hyoid; missing

12/4

49/17

13/7

9/5

9/6

6/4

Hyoid; reduced

44/17

12/6

32/12

42/14

19/8

40/12

* Numerator = Number of foetuses affected; Denominator = Number of litters affected

** Positive control: 250 mg/kg

 

Table 4 - Summary of soft tissue abnormalities

Material

Dose level (mg/kg)

Dam

Number of pups

Description

Sham

0

43004

1

Gastroschisis

 

 

43011

1

Gastroschisis

Aspirin

250.0

43033

3

Encephalomyelocele

 

 

43036

1

Hydrocephalus; Encephalomyelocele

 

 

43038

1

Encephalomyelocele;

anopia

 

 

 

1

Encephalomeningocele;

 

 

43039

1

Encephalomeningocele;

Exophthalmos

Encephalomyelocele

FDA 73-3

 

43043

1

Hydrocephalus;

Gastroschisis

 

 

 

2

Encephalomeningocele;

 

 

43085

1

Gastroschisis

 

 

43132

1

Gastroschisis

 

 

43151

1

Gastroschisis

Conclusions:
Under the conditions of the study, the test material administered to pregnant rats for 10 days up to a dose level of 240 mg/kg bw showed no maternal or developmental toxicity. The NOAEL for both maternal and fetoxicity is > 240 mg/kg bw. Based on this evidence sodium metaphosphate is not considered to be classified as a developmental toxicant according to Regulation (EC) 1272/2008 (EU CLP).

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: no data
Deviations:
not specified
Principles of method if other than guideline:
Adult female albino CD-1 mice were mated with young adult males. Observation of a vaginal sperm plug was considered as Day 0 of gestation. Dosing by oral intubation with a control (Vehicle at level equivalent to group receiving the highest dose or aspirin at 150 mg/kg) or test article in a water suspension (10 mL/kg bw) at 3.7, 17.2, 79.7 and 370.0 mg/kg was carried out daily on Days 6 to 15 of gestation. Observations of body weight, appearence, behaviour, and food consumption were performed. On Day 17 of gestation all dams underwent Caesarean section. Sex, numbers or corpora lutea, implantation sites, resorption sites and live/dead foetuses recorded. Body weights of live pups recorded. Urogenital tract of each dam examined for anatomical normality. All foetuses examined grossly for presence of external congenital abnormalities. One third foetuses of each litter underwent detailed visceral examination and the remaining two thirds were cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.
GLP compliance:
no
Remarks:
Study 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): FDA 73-3 (sodium hexametaphosphate)
- Physical state: Fine white powdered material

Test animals

Species:
mouse
Strain:
other: albino CD-1
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Outbred
- Age at study initiation: No data
- Weight at study initiation: 31 - 34 g
- Fasting period before study: No data
- Housing: Gang housing in disposable plastic cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: No data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 - 26.7
- Humidity (%): 65 - 78

IN-LIFE DATES: 19/7/74 - 23/08/74

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
VEHICLE: Water
- Amount of vehicle (if gavage): 10 mL/kg bodyweight
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
Duration of treatment / exposure:
10 days (Day 6 to Day 15 of gestation)
Frequency of treatment:
Daily
Duration of test:
17 days
No. of animals per sex per dose:
Table 1 Number of animals dosed
Material Dose (mg/kg) Total
Mated Pregnant
Sham 0.0 24 21
Aspirin 150.0 29 19
FDA 73-3 3.7 23 20
17.2 29 20
79.7 30 20
370.0 31 21
Control animals:
yes, sham-exposed
other: positive control: 150 mg/kg aspirin

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Appearence, behaviour, food consumption and weight observed daily.
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
- Time schedule for examinations: Body weights recorded on days 0, 6, 11, 15 and 17.
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 17
- Organs examined: uterus and urogenital tract
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes
Fetal examinations:
- External examinations: Yes: one third per litter
- Soft tissue examinations: Yes: one third per litter
- Skeletal examinations: Yes: two thirds per litter
- Head examinations: Yes: two thirds per litter
Statistics:
No data
Indices:
No data
Historical control data:
No

Results and discussion

Results: maternal animals

Maternal developmental toxicity

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

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
> 370 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no effects observed

Maternal abnormalities

Abnormalities:
no effects observed

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
> 370 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Any other information on results incl. tables

Table 2 - Reproduction data

Dose (mg/kg)

Sham

Aspirin

3.7

17.2

79.7

370.0

Pregnancies

 

 

 

 

 

 

Total No.

21

19

20

20

20

21

Died or aborted (before Day 17)

0

0

0

0

0

1

To term (on Day 17)

21

19

20

20

20

20

Corpora Lutea

 

 

 

 

 

 

Total no.

244

246

257

260

245

249

Average/dam mated

11.1

8.79

11.2

8.97

8.17

8.30

Live litters

 

 

 

 

 

 

Total No.*

21

19

19

20

20

20

Implant Sites

 

 

 

 

 

 

Total No.

226

227

239

245

229

232

Average/dam*

10.8

12.0

12.0

12.3

11.5

11.6

Resorptions

 

 

 

 

 

 

Total No*

10

7

26

10

5

4

Dams with 1 or more sites resorbed

6

6

9

8

3

4

Dams with all sites resorbed

--

--

1

--

--

--

Per cent partial resorptions

28.6

31.6

45.0

40.0

15.0

20.0

Per cent complete resorptions

--

--

5.00

--

--

--

Live foetuses

 

 

 

 

 

 

Total No

215

219

211

232

221

224

Average/dam*

10.2

11.6

10.6

11.6

11.1

11.2

Sex ratio (M/F)

1.11

1.28

1.34

0.99

1.18

0.87

Dead Foetuses

 

 

 

 

 

 

Total No.*

1

1

2

3

3

4

Dams with 1 or more dead

1

1

2

2

3

4

Dams with all dead

--

--

--

--

--

--

Per cent partial dead

4.76

5.26

10.0

10.0

15.0

20.0

Per cent all dead

--

--

--

--

--

--

Average foetus weight (g)

0.88

0.90

0.91

0.89

0.94

0.88

* Includes only those dams examined at term

** Positive control: 150 mg/kg

 

Table 3 - Summary of skeletal findings

Findings

Dose (mg/kg)

Sham

Aspirin

3.7

17.2

79.7

370.0

Live foetuses examined (at term)

151/21

151/19

146/19

161/20

151/20

157/20

Sternebrae

 

 

 

 

 

 

Incomplete oss.

59/18

24/13

56/16

72/18

19/7

28/9

Scrambled

 

 

 

 

 

 

Bipartite

 

3/2

2/2

 

3/3

5/5

Fused

 

 

 

 

 

 

Extra

 

 

 

2/1

 

 

Missing

15/10

15/6

15/10

31/12

14/6

15/8

Other

 

 

 

 

 

 

Ribs

 

 

 

 

 

 

Incomplete oss.

 

 

 

 

 

 

Fused/split

 

 

 

 

 

 

Wavy

 

 

 

 

 

1/1

Less than 12

 

 

 

 

 

 

More than 13

33/17

37/16

25/13

34/12

28/11

34/14

Other

 

 

 

 

 

 

Vertebrae

 

 

 

 

 

 

Incomplete oss.

2/2

3/2

2/2

3/2

9/3

6/5

Scrambled

 

 

 

 

 

 

Fused

 

 

 

 

 

 

Extra ctrs. oss.

 

 

 

 

 

 

Scoliosis

 

 

 

 

 

 

Tail defects

 

 

 

 

 

 

Other

 

 

 

 

 

 

Skull

 

 

 

 

 

 

Incomplete closure

 

 

 

 

 

 

Missing

 

 

 

 

 

 

Craniostosis

 

 

 

 

 

 

Other

 

 

 

 

 

 

Extremities

 

 

 

 

 

 

Incomplete oss.

3/3

2/2

5/5

4/3

6/2

1/1

Missing

 

 

 

 

 

 

Extra

 

 

 

 

 

 

Miscellaneous

 

 

 

 

 

 

Hyoid; missing

32/12

23/13

31/14

34/12

32/12

25/12

Hyoid; reduced

19/14

26/13

16/11

23/12

12/8

32/15

* Numerator = Number of foetuses affected; Denominator = Number of litters affected

** Positive control: 150 mg/kg

 

Table 4 - Summary of soft tissue abnormalities

Material

Dose level (mg/kg)

Dam

Number of pups

Description

FD 73-3

3.7

23073

1

Encephalomeningocele

FD 73-3

370

23178

1

Encephalomeningocele

Applicant's summary and conclusion

Conclusions:
Under the conditions of the study, the test material administered to pregnant mice for 10 days up to a dose level of 370 mg/kg bw showed no maternal or developmental toxicity. The NOAEL for both maternal and fetoxicity is > 370 mg/kg bw. Based on this evidence sodium metaphosphate is not considered to be classified as a developmental toxicant according to Regulation (EC) 1272/2008 (EU CLP).