Reaction mass of Amines, N-tallow alkyltrimethylenedi-, mono(2-ethylhexanoates), Amines, N-tallow alkyltrimethylenedi-, acetates and n-tallow-1,3-diaminopropane ditallate

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 07 July 2004 to 02 February 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

EC Number:

219-145-8

EC Name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

Cas Number:

2372-82-9

Molecular formula:

C18H41N3

IUPAC Name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

Test material form:

other: liquid

Details on test material:

- Physical state: Clear liquid
- Analytical purity: 30.2% (aqueous solution)
- Lot/batch No.: DEGE909567
- Storage condition of test material: At 25 ˚C, in a tightly closed container, protected from heat, direct sunlight and humidity
- Expiry date: February 2005

Test animals

Species:

rabbit

Strain:

Himalayan

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: LPT Laboratory of Pharmacology and Toxicology KG, Hamburg, Germany; branch facility: LPT Löhndorf
- Age at study initiation: 4-5 months
- Weight at study initiation: 2.37-3.26 kg
- Housing: Individually in breeding cages with wire floor (surface of ca. 0.2 m2)
- Diet: Ssniff K-Z V2323, ad libitum
- Water: Ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12/ 12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance preparations were prepared freshly every day. The test substance was suspended in the vehicle to appropriate dose levels.

VEHICLE
- Amount of vehicle (if gavage): 2 mL/kg bw
- Concentration in vehicle: 3, 4.5 and 10 mg/mL

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical determinations of the test substance in aqueous solution and in test substance-carrier were performed.

Details on mating procedure:

- Impregnation procedure: Cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation: Until copulation was ascertained by observation
- Verification of same strain and source of both sexes: Yes
- Proof of pregnancy: Signs of copulation referred to as Day 0 of pregnancy

Duration of treatment / exposure:

Days 6-28 of gestation

Frequency of treatment:

Daily

Duration of test:

Until Day 29 of gestation

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 6, 9 and 20 mg/kg bw/day (pure substance)
Basis:
actual ingested

No. of animals per sex per dose:

20 animals per group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on the results of a dose-range finding study (NOEL of 7.5 mg/kg bw/day, based on reduced food intake and local intolerance reactions).

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once daily
- Cage side observations included: Behavioral changes, reaction to treatment or illness. In addition animals were observed for viability early in the morning and in the afternoon.

BODY WEIGHT: Yes
- Time schedule for examinations: Daily, starting at Day 0 of gestation

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day # 29
- Organs examined: Internal organs

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: Placenta weight was determined.

Fetal examinations:

- External examinations: Yes, all per litter
- Soft tissue examinations: Yes, all per litter
- Skeletal examinations: Yes, all per litter
- Head examinations: Yes, all per litter

Statistics:

The following statistical analysis was used:
For numeric values: Bartlett chi-square test, Dunnett test (for homogeneous variances), Student's t-test (for heterogeneous variances).
For the comparison of classification measurements the Fisher's exact test was employed.
Analytical determinations of Lonzabac in aqueous solution and in test item-carrier were performed.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
Maternal toxicity was indicated at 20 mg/kg/day by the death of two dams, by marginally reduced body weight and food intake during the last days of pregnancy. At 20 mg/kg/day, the body weight was marginally reduced during the last gestion days. There was a reduction in food consumption at 20 mg/kg/day from gestation Day 16 onwards, which was up to 36% below the control value (p = 0.05).
Necropsy revealed an irritation of the gastrointestinal tract in 5 dams and an increased incidence of resorptions.

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

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

Based on:

act. ingr.

Basis for effect level:

other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
- At 20 mg/kg bw/day there was a marginal increased incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity.
- No test substance-related influence was detected on the prenatal foetal development. The incidences of foetal malformations, variations and retardations were similar in all groups. Under the conditions of the study no teratogenic effects (as indicated by the incidences of foetal malformations, variations and retardations) were found up to 20 mg/kg bw/day of the test substance.

Effect levels (fetuses)

1

Dose descriptor:

NOAEL

Effect level:

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

Based on:

act. ingr.

Basis for effect level:

other: embryotoxicity

2

Dose descriptor:

NOAEL

Effect level:

> 20 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Basis for effect level:

other: teratogenicity

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Under the conditions of the study, the test substance was not considered to be a developmental toxicant. The NOAELs for maternal effects and embryotoxicity was set at 9 mg a.i./kg bw/day and the NOAEL for teratogenicity was set at 20 mg a.i./kg bw/day respectively.

Executive summary:

A pre-natal developmental toxicity study was conducted to assess the developmental or teratogenic potential of N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine in Himalayan rabbits according to the OECD Guideline 414 and in compliance with GLP.

 

Groups of 20 pregnant females received the substance by gavage at dose levels of 0, 6, 9 and 20 mg a.i./kg bw/day during Days 6 -28 of gestation. Animals were sacrificed on gestation Day 29.

Maternal toxicity was indicated at 20 mg a.i./kg bw/day by the death of two dams and by marginally reduced body weight and food intake during the last days of pregnancy. Necropsy revealed an irritation of the gastrointestinal tract in 5 dams and an increased incidence of resorptions. Marginal increased incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity was observed at 20 mg a.i./kg bw/day. No substance-related influence was detected on the pre-natal foetal development. The incidences of foetal malformations, variations and retardations were similar in all groups. As a result, the NOAELs for both maternal and embryotoxicity were set at 9 mg a.i./kg bw/day and the NOAEL for teratogenicity was set at 20 mg a.i./kg bw/day.

Under the conditions of the study, the test substance was not considered to be a developmental toxicant.