Manganese Violet

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1973

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

(Uterine weights and corpora lutea were not determined. One third used for visceral examination instead of 50% as recommended in the guideline. Administration only during periods of organogenesis, not until day before pregnancy)

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 3.35, 15.6, 72.3 and 335.0 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. 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

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

7758-16-9

Cas Number:

7758-16-9

IUPAC Name:

7758-16-9

Details on test material:

- Name of test material (as cited in study report): FDA 71-61 (sodium acid pyrophosphate)
- Physical state: Fine white powdered material

Test animals

Species:

mouse

Strain:

CD-1

Remarks:

Albino

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Outbred
- Weight at study initiation: 26.3 - 29.3 g
- Housing: Gang housing in disposable plastic cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum

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

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

3.35 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

15.6 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

72.3 mg/kg bw/day (actual dose received)

Doses / concentrations 4

Dose / conc.:

335 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

Table 1 Number of animals dosed
Material Dose (mg/kg) Total
Mated Pregnant
Sham 0.0 25 25
Aspirin 150.0 24 21
FDA 71-61 3.35 25 23
15.6 25 23
72.3 24 23
335.0 25 22

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: No
- 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

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not examined

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not examined

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

not specified

Changes in number of pregnant:

no effects observed

Other effects:

not examined

Details on maternal toxic effects:

Maternal toxic effects:no effects

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

> 335 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

behaviour (functional findings)

body weight and weight gain

clinical signs

dead fetuses

food consumption and compound intake

histopathology: non-neoplastic

maternal abnormalities

number of abortions

total litter losses by resorption

Results (fetuses)

Fetal body weight changes:

no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

not specified

Changes in litter size and weights:

not specified

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

not specified

Visceral malformations:

no effects observed

Other effects:

not examined

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

> 335 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

reduction in number of live offspring

fetal/pup body weight changes

external malformations

visceral malformations

Remarks on result:

other: developmental toxicity

Fetal abnormalities

Key result

Abnormalities:

not specified

Overall developmental toxicity

Key result

Developmental effects observed:

not specified

Any other information on results incl. tables

Table 2 Reproduction data

Dose (mg/kg)	Sham	Aspirin**	3.35	15.6	72.3	335.0
Pregnancies
Total No.	25	21	23	23	23	22
Died or aborted (before Day 17)	0	0	0	0	0	0
To term (on Day 17)	25	21	23	23	23	22
Live litters
Total No.*	25	20	23	23	23	21
Implant Sites
Total No.	286	254	280	279	265	256
Average/dam*	11.4	12.1	12.2	12.1	11.5	11.6
Resorptions
Total No*	17	37	8	3	10	24
Dams with 1 or more sites resorbed	10	13	6	2	9	9
Dams with all sites resorbed	0	1	0	0	0	1
Per cent partial resorptions	40.0	61.9	26.1	8.70	39.1	40.9
Per cent complete resorptions	--	4.76	--	--	--	4.55
Live foetuses
Total No	264	217	271	274	255	229
Average/dam*	10.6	10.3	11.8	11.9	11.5	10.4
Sex ratio (M/F)	0.96	0.75	0.75	0.73	0.82	0.84
Dead Foetuses
Total No.*	5	0	1	2	0	3
Dams with 1 or more dead	4	--	1	2	--	3
Dams with all dead	0	--	0	0	--	0
Per cent partial dead	16.0	--	4.35	8.70	--	13.6
Per cent all dead	--	--	--	--	--	--
Average foetus weight (g)	0.88	0.84	0.84	0.89	0.86	0.85

* Includes only those dams examined at term

** Positive control: 150 mg/kg

Table 3 Summary of skeletal findings

Findings	Dose (mg/kg)
	Sham	Aspirin**	3.35	15.6	72.3	335.0
Live foetuses examined (at term)	183/25	154/20	188/23	190/23	176/23	159/21
Sternebrae
Incomplete oss.	23/10	34/13	19/11	45/13	33/13	29/9
Scrambled
Bipartite	5/4	6/5	9/7	10/9	5/3	9/6
Fused
Extra		1/1				1/1
Missing	35/14	26/11	27/11	39/8	30/10	14/6
Other
Ribs
Incomplete oss.			3/1		2/1
Fused/split		1/1
Wavy
Less than 12
More than 13	12/7	37/15	34/15	32/14	38/11	31/15
Other
Vertebrae
Incomplete oss.	4/3	9/5	12/4	8/3	4/2	4/2
Scrambled
Fused
Extra ctrs. oss.
Scoliosis
Tail defects
Other
Skull
Incomplete closure			1/1

Missing			1/1
Craniostosis
Other; facial bones, inc				1/1
Extremities
Incomplete oss.	4/3	6/4	12/6	7/3	7/3	4/2
Missing
Extra
Miscellaneous
Hyoid; missing	29/12	35/15	39/15	52/16	46/15	33/13
Hyoid; reduced	17/13	23/13	24/12	18/10	17/13	23/10

* 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
Aspirin	150.0	A 3911	1	Microblepharia Gastroschisis
FDA 71-61	3.35	M 3010	1	Umbilical hernia
FDA 71-61	15.6	M3033	1	Umbilical hernia
		M 3053	1	Hydrocephalus
FDA 71-61	72.3	M3078	1	Hydrocephalus

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 335 mg/kg bw showed no maternal or developmental toxicity. The NOAEL for both maternal and fetotoxicity is > 335 mg/kg bw.

Executive summary:

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 3.35, 15.6, 72.3 and 335.0 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. 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. Under the conditions of the study, the test material administered to pregnant mice for 10 days up to a dose level of 335 mg/kg bw showed no maternal or developmental toxicity. The NOAEL for both maternal and fetotoxicity is > 335 mg/kg bw.