Benzenamine, N-phenyl-, reaction products with 2,4,4-trimethylpentene

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study itself is vaild, but it is only a screening study for developmental toxicity.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 422

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Benzenamine, N-phenyl-, reaction products with 2,4,4-trimethylpentene

EC Number:

270-128-1

EC Name:

Benzenamine, N-phenyl-, reaction products with 2,4,4-trimethylpentene

Cas Number:

68411-46-1

Molecular formula:

C16H19N - C28H43N (main constituents)

IUPAC Name:

Benzenamine, N-phenyl-, reaction products with 2,4,4-trimethylpentene

Test material form:

liquid: viscous

Details on test material:

- Name of test material (as cited in study report): Benzenamine, N-phenyl-,reaction products with 2,4,4-trimethylpentene
- Substance type: UVCB
- Physical state: Clear slightly yellow viscous liquid
- Analytical purity: 100% UVCB
- Purity test date: 2014 , study number 13L00223
- Lot/batch No.: 40401913D
- Expiration date of the lot/batch: 09 February 2016
- Stability under test conditions: stable
- Storage condition of test material: At room temperature in the dark

Test substance handling: Avoid temperatures <10°C, maximum temperature:30°C. In the case of crystallization, heat up the substance (temperature up to 30°C) until a clear solution is obtained
Specific Gravity / Density 0.975 g/cm3 (20°C)
pH 5.1-6.2 (1%(m), 20-25°C) (as suspension)
Stability at higher temperatures Maximum temperature: 30°C

Specific details on test material used for the study:

- Name of test material (as cited in study report): Benzenamine, N-phenyl-,reaction products with 2,4,4-trimethylpentene
- Substance type: UVCB
- Physical state: Clear slightly yellow viscous liquid
- Analytical purity: 100% UVCB
- Purity test date: 2014 , study number 13L00223
- Lot/batch No.: 40401913D
- Expiration date of the lot/batch: 09 February 2016
- Stability under test conditions: stable
- Storage condition of test material: At room temperature in the dark

Test substance handling: Avoid temperatures <10°C, maximum temperature:30°C. In the case of crystallization, heat up the substance (temperature up to 30°C) until a clear solution is obtained
Specific Gravity / Density 0.975 g/cm3 (20°C)
pH 5.1-6.2 (1%(m), 20-25°C) (as suspension)
Stability at higher temperatures Maximum temperature: 30°C

Test animals

Species:

rat

Strain:

other: Crl:WI(Han)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 10-12 weeks
- Weight at study initiation: 353 g (males), 210 g (females)
- Fasting period before study: none
- Housing: groups of 5 animals/sex/cage (premating males and females and postmating males), otherwise single cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%,
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 13 March 2014 To: 1 May 2014

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) are be prepared daily within 6 hours prior to dosing.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Test item is insoluble in water, but soluble in corn oil
- Amount of vehicle (if gavage): 5 mL/kg body weight. Actual dose volumes were calculated according to the latest body weight.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples of formulations were analyzed for homogeneity (highest and lowest concentration) and
accuracy of preparation (all concentrations). Stability in vehicle over 6 hours at room temperature was
also determined (highest and lowest concentration).
The accuracy of preparation was considered acceptable if the mean measured concentrations were
90-110% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was
≤ 10%. Formulations were considered stable if the relative difference before and after storage was
maximally 10%.

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy

Duration of treatment / exposure:

28 days (males), ca 53 days (females)

Frequency of treatment:

daily

Duration of test:

28 days (males), ca 53 days (females)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

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

No. of animals per sex per dose:

10
5 animals/sex/group were selected for functional observations, locomotor activity, clinical pathology, organ weights and histopathology

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Range-finding standy
- Rationale for animal assignment (if not random): Prior to commencement of treatment, by computer-generated random algorithm according to body weight, with all animals within ± 20% of the sex mean.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
The substance caused adverse effects on liver of males and females at dose levels of 75 and 225 mg/kg bw. For details it is referred to the endpoint study records of repeated dose toxicity in IUCLID chapter 7.5.1.

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Maternal abnormalities

Abnormalities:

no effects observed

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Postnatal loss was increased (8 pups in total, versus 0 pups in the control group), but 5 of the 8 pups all came from one dam.
There were not adverse effects regarding other developmental toxicity endpoints at any dose group (eg pup body weight, clinical signs, macroscopic investigation, necropsy).

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

changes in postnatal survival

Remarks on result:

other: Increase in postnatal pup mortality at 225 mg/kg bw., mostly in one animal and unlikely to be a true effect

Fetal abnormalities

Abnormalities:

not examined

Overall developmental toxicity

Developmental effects observed:

yes

Lowest effective dose / conc.:

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

Treatment related:

yes

Relation to maternal toxicity:

developmental effects as a secondary non-specific consequence of maternal toxicity effects

Dose response relationship:

yes