[No public or meaningful name is available]

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From June 19, 1979 to June 12, 1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Reference 2

Reference Type:

study report

Title:

Unnamed

Year:

1980

Report date:

1980

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

No macroscopic or microscopic examination of sacrificed dams

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N,N,N-trimethyl-C12-14 (even numbered)-alkyl-1-aminium chloride

Molecular formula:

C15H34Cl1N1 (representative molecular formula of C12 chain)

IUPAC Name:

N,N,N-trimethyl-C12-14 (even numbered)-alkyl-1-aminium chloride

Constituent 2

Reference substance name:

Alcohols, C14-15, ethoxylated

EC Number:

614-831-7

Cas Number:

68951-67-7

IUPAC Name:

Alcohols, C14-15, ethoxylated

Test material form:

other: White-cream, glossy paste

Specific details on test material used for the study:

Test substance: C12-14 TMAC
Purity: 35% C12-14 TMAC in C14-15 Pareth-7
Appreance: White cream, glossy paste

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Animal supplier source: Accredited breeder - Oury, Domaine De la Chesnaie, 45230 Chatilloon-Coligny
Body weight: 2.7 to 2.9 kg
Sex: Female
Pre-trail period: The animal were maintained under conditions as close as possible to those of the trial for one month pre-trial and were vaccinated against myxomatosis on May 12, 1979. The male rabbits used for mating were supplied from the same breeder and placed under the same experimental conditions as the females
Housing: Animals were placed in individual caging (70×50×30 cm) with plastic perforated floor
Temperature: 21ºC ± 10ºC
Relative humidity: 50 ± 10%
Air changes: 10 times/hour
Allocation to the groups: 13 animals each were grouped to control, test group 1, test group 2 and test group 3.
Tap water: ad libitum

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

The first mating started on September 04, 1979.

Duration of treatment / exposure:

The day of mating was considered as day 0 and the animals were dosed daily from day 6 to day 18 inclusive (for 13 consecutive days).

Frequency of treatment:

once a day

Duration of test:

28 d

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

2 mg/kg bw/day

Remarks:

0.35 mg a.i./kg bw/day

Doses / concentrations 2

Dose / conc.:

8 mg/kg bw/day

Remarks:

1.4 mg a.i./kg bw/day

Doses / concentrations 3

Dose / conc.:

24 mg/kg bw/day

Remarks:

8.4 mg a.i./kg bw/day

No. of animals per sex per dose:

13 female rabbits per dose

Control animals:

yes

Details on study design:

Preliminary study
Animal details: 3 female New Zealand rabbits for each dose group, weighing about 3 kg
Route of administration: Oral by stomach tube
Dose levels (mg/kg bw/day): 25 mg/kg bw/day (Test group 1), 50 mg/kg bw/day (Test group 2), 100 mg/kg bw/day (Test group 3), 200 mg/kg bw/day (Test group 4), 400 mg/kg bw/day (Test group 5)
Test substance was administered as an aqueous solution at following concentration:
Test group 1: 1.25 %
Test group 2: 2.50 %
Test group 3: 5 %
Test group 4: 10 %
Test group 5: 20 %
The solution were prepared every 3rd or 4th d and stored at 4ºC. 2 mL/kg bw was administered for each group. The preliminary test results obtained indicated that the test substance was toxic at the dose levels of 100, 200 and 400 mg/kg bw/day by oral route. 50 mg/kg did seem to be the threshold of an indirect embryotoxic action.

Main study
Based on preliminary testing results, following dose levels were selected for main study:
Test group 1: 1 mg/kg bw/day
Test group 2: 5 mg/kg bw/day
Test group 3: 25 mg/kg bw/day

Examinations

Maternal examinations:

- Behaviour and clinical symptoms: daily
- Body weight: every 3 days during pregnancy
- Food consumption: daily
- Number of corpora lutea, implantations, resorptions

Ovaries and uterine content:

Staining of implantation sites with ammonium sulphide
- Number of corpora lutea
- Number of implantations
- Number of absorptions

Fetal examinations:

- Number of viable and dead foetuses
- Macroscopic external observations
- Individual weights
- Sex
- 2/3 of foetuses stained with alizarin for skeletal examination
- 1/3 of foetuses immersed on Bouin: sections cut for examination of visceral abnormalities (method adapted from Wilson technique)

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

Description (incidence and severity):

No difference of growth curve of treated groups compared to controls was observed.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No difference of growth curve of treated groups compared to controls was observed.

Food efficiency:

not examined

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):

not examined

Immunological findings:

not examined

Gross pathological findings:

not examined

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Details on results:

No mortality, no difference of growth curve of treated groups compared to controls, no difference in food consumption.

Maternal developmental toxicity

Number of abortions:

no effects observed

Early or late resorptions:

no effects observed

Description (incidence and severity):

The rate of resorptions was considered low and comparable in all groups.

Dead fetuses:

no effects observed

Details on maternal toxic effects:

There was a higher number of non-pregnant females in group I. The rate of resorptions is considered low, and comparable in all groups. No histopathology was performed on the sacrificed dams end of term.

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 24 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: No effects were observed at highest test dose

Remarks on result:

other:

Remarks:

NOAEL 8.4 mg a.i./kg bw/day

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

External malformations:

no effects observed

Description (incidence and severity):

In the high dose group, one foetus showed a torsion of the fore limbs. No other external abnormality was observed in any of the dose groups.

Skeletal malformations:

no effects observed

Description (incidence and severity):

Incidence in treated groups were comparable to controls.

Visceral malformations:

no effects observed

Description (incidence and severity):

- Dilated and haemorrhagic cerebral ventricles in one foetus of control group
- Left hydro-ureter in one foetus of low dose group
- Retinal folds in region of optic nerve in 13 foetuses: Control: 3 (11%); low dose: 2 (9%); mid dose: 4 (14%); and high dose: 4 (16%). These were considered to be caused by fixation artefact.

Details on embryotoxic / teratogenic effects:

No foetal mortality was observed.

External abnormalities:
In the high dose group, one foetus showed a torsion of the fore limbs. No other external abnormality was observed in any of the dose groups.

Visceral abnormalities:
- dilated and haemorrhagic cerebral ventricles in one foetus of control group
- left hydro-ureter in one foetus of low dose group.
- retinal folds in region of optic nerve in 13 foetuses: Control: 3 (11%); low dose: 2 (9%); mid dose: 4 (14%); and high dose: 4 (16%). These were considered to be caused by fixation artefact.

Skeletal abnormalities: See table: Incidence in treated groups are comparable to controls.

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 24 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects were observed at highest test dose

Remarks on result:

other: NOAEL: 8.4 mg a.i./kgbw/day

Fetal abnormalities

Key result

Abnormalities:

no effects observed

Overall developmental toxicity

Key result

Developmental effects observed:

no

Any other information on results incl. tables

Maternal toxic Effects

 

Preliminary study

25, 50, 100, 200, and 400 mg test material /kg bw/day using 3 animals per dose group.

 

Mortality

Dose of test substance (mg/kg bw/day)	25	50	100	200	400
mortality	1/3 (day 26)	1/3 (day 18)	2/3 (day 18,26)	3/3 (all day 10)	3/3 (all day 7)

At 50 and 100 mg groups, a clear fall in body weight was observed between days 12 and 18. At 25 mg, two of the three females showed anorexia. From first day of treatment food consumption was clearly decreased at 50 and 100 mg/kg.

 

Final study

No mortality, no difference of growth curve of treated groups compared to controls, no difference in food consumption. There was a higher number of non-pregnant females in group I. The rate of resorptions is considered low, and comparable in all groups. No histopathology was performed on the sacrificed dams end of term.

 

Teratogenic/embryotoxic effects

 

Preliminary study on 25, 50, 100, 200, and 400 mg test material/kg bw/day using 3 animals per dose group. At 50 mg, the two females showed resorption sites. The weight of foetuses of the 50 mg group were low. Macroscopic examination, number and mean weight of foetuses of the 25 mg group showed no embryotoxic effect. 50 mg/kg/day seems to be the threshold for an indirect embryotoxic action.

 

Final study

No foetal mortality was observed.

External abnormalities
In the high dose group, one foetus showed a torsion of the fore limbs. No other external abnormality was observed in any of the dose groups.

Visceral abnormalities

- Dilated and haemorrhagic cerebral ventricles in one foetus of control group

- left hydro-ureter in one foetus of low dose group.

- Retinal folds in region of optic nerve in 13 foetuses: Control: 3 (11%); low dose: 2 (9%); mid dose: 4 (14%); and high dose: 4 (16%). These were considered to be caused by fixation artefact.
Skeletal abnormalities: Incidence in treated groups are comparable to controls.

Table for Maternal effects (separate data for all dosage groups) 

Maternal effects
Parameter	control data		low dose	medium dose	high dose	dose-response + / -
	historical	study
Number of dams examined		13	14	13	13
Clinical findings during application of test substance		no	no	no	no
Mortality of dams state %		0	0	0	0
Abortions		0	0	0	0
Body weight (day 28) g gain day 0-28 (end of test) g, gain day 6-18 (treatment) g,		3468 541 205	3313 638 206	3450 645 246	3369 650 231	-
Food consumption (g/day/dam) day 0-5 day 6-18 (treatment) day 19-29		165 152 111	148 139 118	148 143 124	157 152 134	-
Water consumption if test substance is applied with drinking water		not recorded	not recorded	not recorded	not recorded
Pregnancies pregnancy rate or %		11 85%	8 57%	11 85%	13 100%	+/-
Necropsy findings in dams dead before end of test		none died	none died	none died	none died

 Table for developmental effects (separate data for all dosage groups)

Litter response (Caesarean section data)
Parameter	control data		low dose	medium dose	high dose	dose-response + / -
	historical	study
Number full-term pregnant females		11	8	11	13	+/-
Corpora lutea number per pregnant female		98 8.91	73 9.13	97 8.82	104 8.00	+/-
Implantations number per pregnant female		84 7.64	71 8.87	85 7.73	98 7.54	-
Resorptions number per pregnant female		1 0.09	4 0.50	2 0.18	7 0.54	-
total number of foetuses number per pregnant female		83 7.55	67 8.37	83 7.55	91 7.00	+/-
pre-implantation loss state %		14%	3%	12%	6%	-
post-implantation loss state %		1%	6%	2%	7%	-
total number of litters		11	8	11	13	+/-
fetuses / litter		7.55	8.37	7.55	7.00	+/-
live fetuses / litter state ratio		7.55	8.37	7.55	7.00	+/-
dead fetuses / litter state ratio		0	0	0	0
fetus weight (mean) [g]		33.63	31.73	34.21	35.17	+/-
placenta weight (mean) [g]		not recorded	not recorded	not recorded	not recorded
crown-rump length (mean) [mm]		not recorded	not recorded	not recorded	not recorded
Fetal sex ratio [state ratio m/f]		40/38	26/34	39/39	48/34	+/-

 Table for developmental effects (separate data for all dosage groups)

Examination of the foetuses
Parameter	control data		low dose	medium dose	high dose	dose-response + / -
	historical	study
External abnormalities [%]		1 3.7%	0	0	0	-
Skeletal abnormal ossifications 12^thpair complete [%] 13ribs left side [%] 13ribs right side [%] 13^thpair complete [%]		47.1% 3.9% 5.9% 43.1%	8.9% 11.1%	30.9% 10.9% 10.9% 47.3%	45.0% 10.0% 6.7% 38.3%	-
incomplete ossifications sternebrae [%]		32 63%	29 64%	30 55%	36 60%	-
incomplete ossifications skull [%]		23 45%	9 20%	11 20%	15 25%	-
missing ossifications sternebrae [%]		14 27%	11 24%	12 22%	21 35%
abnormal ossifications		1 2%		1 1.8%		-
Visceral abnormalities [%]		1 3.7%	1 4.5%	0	0	-

Applicant's summary and conclusion

Conclusions:

Under study conditions, the NOAEL of test substance for maternal and embryotoxic effects/teratogenicty was determined to be 24 mg/kg bw/day (i.e., equivalent to 8.4 mg a.i./kg bw/day)

Executive summary:

A study was conducted to determine the teratogenicity of test substance, C12-14 TMAC (active ingredient 35%), according to the method comparable to OECD 414. During gestation Days 6 -18, four groups of 13 female New Zealand White rabbits were dosed by stomach tube at dose levels of 0, 2, 8 and 24 mg/kg bw/day of test substance (35% C12 -14 -TMAC in dobanol 45E7 (i.e., C14 -15 AE7)), in 2 mL/kg bw/day daily prepared solutions in water. Animals were sacrificed on Day 29. Examinations included daily observations of behaviour and clinical symptoms, daily food consumption, and body weight every three days. Upon sacrifice, number of corpora lutea, implantations and resorptions were examined. For foetuses, number of viable and dead, macroscopic external examinations, sex, individual weights, skeletal examinations (2/3), and visceral abnormalities (1/3) were conducted. A preliminary study was undertaken under similar conditions, applying 5 dose groups: 0, 25, 50, 100, 200 and 400 mg/kg bw/day using three animals per dose group. At 50 mg, the two females showed resorption sites. The weight of foetuses of the 50 mg group were low. Macroscopic examination, number and mean weight of foetuses of the 25 mg group showed no embryotoxic effect. 50 mg/kg/day did seem to be the threshold for an indirect embryotoxic action. In the final study, no signs of maternal toxicity was observed. Also no foetal mortality was observed. Numbers of corpora lutea and resorptions were comparable. The number of pregnancies at the low dose group did seem to be diminished, but this was considered an incidental observation. There was no change in frequency of external, visceral and skeletal observations. No maternal toxic or embryotoxic/teratogenic effects were seen up to hightest tested dose of 24 mg/kg bw/day (i.e., equivalent to 8.4 mg a.i./kg bw/day). Under study conditions, the NOAEL of test substance for maternal and embryotoxic effects/teratogenicty in rabbits was determined to be 24 mg/kg bw/day (i.e., equivalent to 8.4 mg a.i./kg bw/day) (Fave 1980).