4,4'-benzylidenedianiline

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 26 November 2009 to 30 December 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

2001

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Version / remarks:

No. 440/2008

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-benzylidenedianiline

EC Number:

210-038-1

EC Name:

4,4'-benzylidenedianiline

Cas Number:

603-40-7

Molecular formula:

C19H18N2

IUPAC Name:

4-[(4-aminophenyl)(phenyl)methyl]aniline

Test material form:

solid

Details on test material:

Identification: DATIPH
Physical form: Solid
Solubility in water: < 1 g/L
Colour: yellow brown
CAS no.: 603-40-7
Chemical name: 4,4’-Benzylidenedianiline
Molecular formula: C19H18N2
Test substance storage: At room temperature protected from light
Stability: Stable under storage conditions
Expiry date: 31 October 2010

Specific details on test material used for the study:

Source of test material:
Sponsor (identification DATIPH)
- Description: brown solid lumps 09 November 2009
- Date received: 09 November 2009
- Storage conditions: approximately 4°C in the dark

The integrity of supplied data relating to the identity, purity and stability of the test material is the responsibility of the Sponsor.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: For the purpose of the study the test material was freshly prepared, as required, as a suspension in arachis oil BP. Arachis oil BP was used because the test material did not dissolve/suspend in distilled water.
- Determination by analysis of the concentration, homogeneity and stability of the test material preparations was not appropriate because it was not specified in the Study Plan and is nota requirement of the Test Guideline.

Test animals

Species:

rat

Strain:

Wistar

Remarks:

HsdRccHan®™:WIST®™

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: HsdRccHan®™:WIST®™
- Females nulliparous and non-pregnant: yes
- Age at study initiation: eight to twelve weeks old
- Weight at study initiation:
- Fasting period: overnight fast immediately before dosing and for approximately three to four hours after dosing
- Housing: groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes
- Diet (ad libitum): 2014 Teklad Global Rodent diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK
- Water (ad libitum): 2014 Teklad Global Rodent diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06:00 to 18:00) and twelve hours darkness
- The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 200mg/mL
- Amount of vehicle (if gavage): 10ml/kg
- Justification for choice of vehicle: Test material was not soluble in water

MAXIMUM DOSE VOLUME APPLIED: 2000mg/kg

- Rationale for the selection of the starting dose: ln the absence of data regarding the toxicity of the test material, 300 mg/kg was chosen as the starting dose

VEHICLE
- The volume administered: calculated according to the fasted bodyweight at the time of dosing for each animal. Treatment was sequential.
- Justification for choice of vehicle: Test material was not soluble in water

Doses:

300mg/kg, 2000mg/kg

No. of animals per sex per dose:

1 (+4 additional animals for the 2000mg/kg dose)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 30min, 1h, 2h, 4h and daily for 14 days. Morbidity and mortality checks were made twice daily. lndividual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes, the animais were killed by cervical dislocation and subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities.
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There was no mortality.

Clinical signs:

At dose level of 300mg/kg, no signs of systemic toxicity were noted during the observation period.
At dose level of 2000mg/kg, signs of systemic toxicity noted in one animal were hunched posture, ataxia and exophthalmos. No other signs of systemic toxicity were noted (Table 1).

Body weight:

At dose level of 300mg/kg, the animal showed expected gains in bodyweight over the observation period.
At dose level of 2000mg/kg, animals showed expected gains in bodyweight except for two animals which showed bodyweight loss or no gain in bodyweight during the first week but expected gain in bodyweight during the second week.

Other findings:

No abnormalities were noted at necropsy.

Any other information on results incl. tables

Table 1. lndividual Clinical Observations and Mortality Data

Dose Levelmg/kg	AnimalNumber and Sex	Effects Noted After Dosing (Hours)				Effects Noted During Period After Dosing (Days)
		0.5	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	2-0 Female	0	0	0	HAB	HAB	H	H	0	0	0	0	0	0	0	0	0	0	0
	3-0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-1 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-2 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-3 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
300	1 -0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

0 = No signs of systemic toxicity; H = Hunched posture; A= Ataxia; B=Exophthalamos

Table 2. lndividual Bodyweights and Bodyweight Changes

Dose Level mg/kg	Animal Number and Sex	Bodyweight (g) at Day			Bodyweight Gain (g) During Week
		0	7	14	1	2
2000	2-0 Female	160	161	180	1	19
	3-0 Female	177	178	186	1	8
	3-1 Female	161	160	166	-1	6
	3-2 Female	176	176	186	0	10
	3-3 Female	185	186	189	1	3
300	1-0 Female	175	186	196	11	10

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat is estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System - Category 5).

Executive summary:

The study was performed to assess the acute oral toxicity of the 4,4'-Benzylidenedianiline in the Wistar strain rat according to the OECD guideline 420 and method B1 bis of the Commission Regulation (EC) No. 440/2008. 4,4'-Benzylidenedianiline was administered at doses of 300mg/kg and 2000mg/kg. Signs of systemic toxicity noted in one animal treated at a dose level of 2000 mg/kg were hunched posture, ataxia and exophthalmos. No other signs of systemic toxicity or anormalities at the autopsy were noted and there were no deaths. The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat is estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System -  Category 5).