Reaction mass of N1,N1'-(methanediyldibenzene-4,1 -diyl)diaryldiamine and 2-{4-[(4-aminophenyl)amino]benzyl}-N4-(4-{4-[(4-aminophenyl)amino]benzyl}phenyl)aryldiamine

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 September 2008 - 07 November 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study; well documented study report

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid

Details on test material:

- Appearance/physical state: Black lumps
- Storage conditions: Room temperature in the dark

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
-Source: Charles River Deutschland, Sulzfeld, Germany.
-Age at study initiation: approx. 8~10 weeks
-Weight at study initiation: Body weight variation did notexceed +/-20% of the sex mean.
-Fasting period before study: Food was withheld overnight (for a maximum of 20 hours) prior to dosing until 3-4 hours after administration of the test substance.
-Housing: Group housing of 3 animals per cage in labeled Macrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)..
-Diet: Free access to pelleted rodent diet (SM RIM-Z from SSNIFFB Spezialdiaten GmbH, Soest, Germany).
-Water: free access to tap-water.
-Acclimation period: Acclimatization period was at least 5 days before start of treatment under laboratory conditions.
ENVIRONMENTAL CONDITIONS
-Temperature (C): 19.5 -21.3 C
-Humidity (%): 30-70% (actual range: 37 -74%)
-Air changes: 15 air changes per hr
-Photoperiod: 12 hours artificial fluorescent light and 12 hours darkness per day.

IN-LIFE DATES: Start: 23 September 2008; Completion: 07 November 2008.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30, and 200 mg/ml
- Amount of vehicle (if gavage): 10 ml/kg bw
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at NOTOX and on test substance data supplied by the sponsor.
- Lot/batch no. (if required): No data available.
- Purity: No data available.

MAXIMUM DOSE VOLUME APPLIED: 10mL/Kg
DOSAGE PREPARATION (if unusual): the formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: the toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

12 Females, each dose group consisted of 3 animals.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:

-Motility/viability: twice daily. The time of death was recorded as precisely as possible.

-Body weights: Days 1 (pre-administration), 8 and 15 and at death.

-Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.


- Necropsy of survivors performed: yes

- Other examinations performed: None.

Statistics:

No statistical analysis was performed.

Results and discussion

Preliminary study:

Not applicable

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

500 mg/kg bw

Mortality:

Five females in the high dose group were found dead on Days 9. 12 or 13. No further mortality occurred.

Clinical signs:

Clinical signs observed during the study period were as follows:
2000 mg/kg group: lethargy, hunched posture, watery discharge of the eyes, uncoordinated movements, ptosis, piloerection, lean appearance and/or hypothermia were noted in all animals during the observation period.
300 mg/kg group: hunched posture and piloerection were noted in all animals between
Days 1 and 7.

Body weight:

The mean body weight gain shown by the animals at 300 mg/kg over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

Emaciation, dark red discoloration of the thymus and reduced size of the thymus was noted in one surviving animal at 2000 mg/kg. One animal, which was found dead, showed irregular surface of the forestomach. All animals found dead showed beginning or advanced autolysis.
No abnormalities were found at macroscopic post mortem examination of the animals at 300 mg/kg.

Other findings:

- Organ weights: No data available.
- Histopathology: No data available.
- Potential target organs: No data available.
- Other observations: No data available.

Any other information on results incl. tables

Results are shown in the attachments.

Table 1: Mortality

Table 2: Clinical signs

Table 3: Body weights

Table 4: Macroscopic findings

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Migrated information

Conclusions:

The oral LD50 value of the test substance in Wistar rats was established to be within the range of 300- 2000 mg/kg body weight. According to the OECD 423 test guideline the LD50 cut-off value was considered to be 500mg/kg body weight.

Executive summary:

In an acute oral toxicity study, the oral LD50value of the test substance in Wistar rats was established to be within the range of 300- 2000 mg/kg body weight. According to the OECD 423 test guideline the LD50 cut-off value was considered to be 500mg/kg body weight.