Trimethyl[2-(phosphonooxy)ethyl]ammonium chloride, calcium salt (1:1)

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1973-1974

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:

1974

Report date:

1974

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

only one-third of each litter were examined

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 002

EC Number:

600-059-8

Cas Number:

10031-30-8

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Fine white crystalline material.

Test animals

Species:

mouse

Strain:

CD-1

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: adult
- Weight at study initiation: 30 ± 1 g
- Fasting period before study: no data
- 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

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

Virgin adult female albino outbred mice were mated with young adult males, and observation of the vaginal sperm plug was considered Day 0 of gestation.

Duration of treatment / exposure:

10 days (from day 6 to day 15 of gestation)

Frequency of treatment:

Daily

Duration of test:

17 days

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

4.65 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

21.6 mg/kg bw/day

Doses / concentrations 3

Dose / conc.:

100 mg/kg bw/day

Doses / concentrations 4

Dose / conc.:

465 mg/kg bw/day

No. of animals per sex per dose:

Sham: 22 animals
Positive control: 20 animals
4.65 mg/kg: 24 animals
21.6 mg/kg: 19 animals
100 mg/kg: 22 animals
465 mg/kg: 23 animals

Control animals:

yes, concurrent vehicle

other: Aspirin 150 mg/kg (positive control)

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: appearance and behaviour.

BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 11, 15, and 17 of gestation

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 17
- Organs examined: sternebrae, ribs, vertebrae, skull, extremities

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Number of dead fetuses: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: one-third per litter
- Skeletal 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 specified

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not specified

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

effects observed, non-treatment-related

Early or late resorptions:

not specified

Dead fetuses:

effects observed, non-treatment-related

Changes in pregnancy duration:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed

Changes in number of pregnant:

not specified

Other effects:

not specified

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

> 465 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

dead fetuses

early or late resorptions

number of abortions

pre and post implantation loss

total litter losses by resorption

Results (fetuses)

Fetal body weight changes:

no effects observed

Reduction in number of live offspring:

effects observed, non-treatment-related

Changes in sex ratio:

not specified

Changes in litter size and weights:

not specified

Changes in postnatal survival:

not specified

External malformations:

no effects observed

Skeletal malformations:

effects observed, non-treatment-related

Visceral malformations:

effects observed, non-treatment-related

Other effects:

not specified

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

> 465 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

external malformations

skeletal malformations

visceral malformations

Overall developmental toxicity

Key result

Developmental effects observed:

no

Any other information on results incl. tables

Table 1. Fate summary

Material	Dose** mg/kg	Total		Surviving at Term
		Mated	Pregnant	Total	Pregnant^1
Sham	0	25	22	25	22
Aspirin*	150	25	20	24	19
Monocalcium phosphate	4.65	25	24	25	24
Monocalcium phosphate	21.6	26	19	26	19
Monocalcium phosphate	100	23	22	23	22
Monocalcium phosphate	465	25	23	25	23

* Positive Control: 150 mg/kg

** Administered as a water solution (10 ml per kg of body weight)

¹includes all dams examined at term

 

Table 2. Reproduction data.

 

	Sham	Aspirin**	Monocalcium phosphate 4.65 mg/kg	Monocalcium phosphate 21.6 mg/kg	Monocalcium phosphate 100 mg/kg	Monocalcium phosphate 465 mg/kg
Pregnancies
Total No.	22	20	24	19	22	23
Died or aborted (before Day 17)	0	1	0	0	0	0
To term (on Day 17)	22	19	24	19	22	23
Live Litters
Total No.*	22	19	24	19	22	22
Implant sites
Total No.	261	224	283	225	244	265
Average/dam*	11.9	11.8	11.8	11.8	11.1	11.5
Resorptions
Total No.*	8	20	19	4	12	28
Dams with 1 or more sites resorbed	7	9	13	3	10	12
Dams with all sites resorbed	--	--	--	--	--	1
Per cent partial resorptions	31.8	47.4	54.2	15.8	45.5	52.2
Per cent complete resorptions	--	--	--	--	--	4.35
Live Fetuses
Total No.	252	201	261	221	230	233
Average/dam*	11.5	10.6	10.9	11.6	10.5	10.1
Sex ratio (M/F)	1.02	0.88	0.78	0.92	1.13	0.93
Dead Fetuses
Total*	1	3	--	--	2	4
Dams with 1 or more dead	1	3	--	--	2	3
Dams with all dead	--	--	--	--	--	--
Per cent partial dead	4.55	15.8	--	--	9.09	13.0
Per cent all dead	--	--	--	--	--	--
Average Fetus Weight, g	0.84	0.81	0.88	0.87	0.82	0.85

* Includes only those dams examined at term

** Positive control: 150 mg/kg

 

Table 3. Summary of skeletal findings.

 

	Sham	Aspirin**	Monocalcium phosphate 4.65 mg/kg	Monocalcium phosphate 21.6 mg/kg	Monocalcium phosphate 100 mg/kg	Monocalcium phosphate 465 mg/kg
Live Fetuses Examined (at term)	179/22	141/19	186/24	155/19	162/22	164/22
Sternebrae
Incomplete oss.	10/8	31/10	18/9	27/15	22/10	28/15
Scrambled
Bipartite	8/7	3/3	11/8	9/7	10/8	9/6
Fused
Extra		6/3
Missing	23/10	28/11	13/9	13/6	32/14	23/7
Other
Ribs
Incomplete oss.						9/4
Fused/split		2/2
Wavy		1/1	2/2
Less than 12	1/1		1/1			1/1
Less than 13	48/18	30/12	42/18	18/11	32/15	34/17
Other
Vertebrae
Incomplete oss.	9/6	9/3	2/2	1/1	11/4	13/4
Scrambled
Fused
Extra ctrs. Oss.
Scoliosis
Tail defects
Other
Skull
Incomplete closure		2/2
Missing			1/1
Craniostosis
Other, facial bones, inc.						2/1
Extremities
Incomplete oss.	7/5	5/3	2/2		10/5	14/4
Missing
Extra
Miscellaneous
Hyoid; missing	37/16	33/11	13/15	22/12	52/15	33/13
Hyoid; reduced	17/11	25/12	17/11	21/14	15/11	27/14

* Number of fetuses affected/number of litters

** Positive control: 150 mg/kg

 

Table 3-a. Summary of soft tissue abnormalities.

 

Material	Dose level (mg/kg)	Dam	Number of pups	Description
Aspirin*	150	A 6068	1	Cleft palate; gastroschisis

*Positive control: 150 mg/kg

Table 4. Average body weights of pregnant dams.

 

Material	Dose level (mg/kg)	Day 0	Day 6	Day 11	Day 15	Day 17*
Sham	0	30.5	32.7	37.1	44.1	51.3 (22)
Aspirin**	150	30.0	32.2	33.7	40.6	47.0 (19)
Monocalcium phosphate	4.65	29.3	32.3	36.5	43.7	50.2 (24)
Monocalcium phosphate	21.6	29.8	32.2	36.4	44.3	50.8 (19)
Monocalcium phosphate	100	30.6	33.5	36.7	44.0	48.1 (22)
Monocalcium phosphate	465	29.2	32.3	35.1	42.8	47.8 (23)

* Number of surviving dams in parenthesis (c.f. Table 1)

** Positive control: 150 mg/kg

Applicant's summary and conclusion

Conclusions:

The test item up to 465 mg/kg bw/day did not exhibit any maternal toxicity, fetal survival, teratogenicity in female mice CD-1. Therefore, the NOAEL in mice was higher than 465 mg/kg bw/d.

Executive summary:

A study was conducted to assess the teratogenic potential of the oral administration of monocalcium phosphate in mice by a method similar to OECD 414. Starting from day 0 of gestation, female mated CD-1 mice were orally administered 0 (sham), 4.65, 21.6, 100 or 465 mg/kg bw for 10 consecutive days. A positive control group was administered in parallel with 150 mg/kg of aspirin. Maternal examinations had no clearly discernible effect on nidation or on maternal or fetal survival. Resorptions and dead fetuses were counted. One-third of the fetuses from each litter underwent detailed visceral examinations. The remaining two-thirds were examined for skeletal defects. Based on the test results, the test item up to 465 mg/kg bw/day did not exhibit any maternal toxicity, fetal survival, teratogenicity in female mice CD-1. Therefore, the NOAEL for both maternal and developmental toxicity in mice was higher than 465 mg/kg bw/d.