Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2002-01-29 to 2002-02-14 (experimental phase)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study without deviations, not performed under GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
no
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): a reaction product of 1,1'-benzene-1 ,3-diyI)bis(methylamine) and styrene (other name: MXDA/SM adduct)
- Physical state: Pale yellow liquid (degree of viscosity: 6.8 mPa · s/25 °C)
- Analytical purity: not reported
- Impurities (identity and concentrations): not reported
- Composition of test material, percentage of components: components given in the study report still were based on information retrieved from old GC analytical data (for details see chapter 1.2)
Component a: 54.9 wt%
Component b: 4.7 wt%
Component c: 36. 4wt%
Component d: 4.0 wt%
- Purity test date: not reported
- Lot/batch No.: not reported
- Stability under test conditions: not reported
- Storage condition of test material: stored in a refrigerator until use

Test animals

Species:
rat
Strain:
other: Crj: CD(SD)IGS, SPF
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan Co., Ltd.
- Age at study initiation: 5 weeks
- Weight at study initiation: 113 ± 2.5 g (males), 97.0 ± 3.4 g (females)
- Fasting period before study: Rats were fasted overnight. Feeding was restarted three to four hours after dosing.
- Housing: in groups of six to ten in suspended metal wire cages on a rat cage rack equipped with a feed water supply system (Tokiwa Scientific Instrument Co., Ltd.)
- Diet (e.g. ad libitum): free access to a rat pellet diet MF (Oriental Yeast Co., Ltd.)
- Water (e.g. ad libitum): free access to mains water
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 2
- Humidity (%): 55 ± 6
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: from 2002-01-29 to 2002-02-14

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.25, 0.5, 2, 5 and 20 (w/v)% olive oil solutions
- Amount of vehicle (if gavage): 1.0 mL per 100 g body weight
- Justification for choice of vehicle: not reported

MAXIMUM DOSE VOLUME APPLIED: 1.0 mL per 100 g body weight

DOSAGE PREPARATION (if unusual): A prescribed amount of test material was weighed out and diluted with olive oil in order to formulate 0.25 to 20 (w/v)% olive oil solutions as test solutions.
Doses:
25, 50, 200, 500 and 2000 mg/kg
No. of animals per sex per dose:
6 animals per sex and dose
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observed daily for mortality and clinical signs, weighed immediately before dosing and on Days 2, 7, and 14
- Necropsy of survivors performed: yes
- Other examinations performed: mortality, clinical signs, body weight

Results and discussion

Effect levelsopen allclose all
Sex:
male/female
Dose descriptor:
discriminating dose
Effect level:
500 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: This value is calculated as the geometric mean of the highest test dose without mortalities (500 mg/kg) and the first one with mortality in all test animals.
Mortality:
Male: 25 mg/kg bw; Number of animals: 6; Number of deaths: 0
Male: 50 mg/kg bw; Number of animals: 6; Number of deaths: 0
Male: 200 mg/kg bw; Number of animals: 6; Number of deaths: 0
Male: 500 mg/kg bw; Number of animals: 6; Number of deaths: 0
Male: 2000 mg/kg bw; Number of animals: 6; Number of deaths: 6
Female: 25 mg/kg bw; Number of animals: 6; Number of deaths: 0
Female: 50 mg/kg bw; Number of animals: 6; Number of deaths: 0
Female: 200 mg/kg bw; Number of animals: 6; Number of deaths: 0
Female: 500 mg/kg bw; Number of animals: 6; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 6; Number of deaths: 6
Clinical signs:
Males from the 25, 50 and 200 mg/kg dose groups showed no abnormalities. In the 500 mg/kg dose group, all of the six males showed body descent, but five of them recovered two hours after treatment. One animal from the dose group showed body descent and abnormal gait, but recovered five hours after dosing. Diarrhea was noted in three males from the same dose group. Loss of fur was noted in four males on the day following the treatment day, but no abnormalities were seen on Day 3 or later.
All of the males from the 2000 mg/kg dose group showed body descent within 30 minutes after dosing and one male also showed tremor. All of the six males were prostrated and died 40 minutes to three hours after dosing, showing jumping and struggling.
No abnormalities were observed in females dosed at 26, 50, and 200 mg/kg. In the 500 mg/kg dose group, all of the six females showed body descent 30 minutes after dosing and one of them also showed squatting. All of them recovered two hours after dosing, but two of them showed diarrhea five hours after treatment. Five females showed loss of fur on the day following the treatment day, but no abnormalities on Day 3 or later.
All of the six females from the 2000 mg/kg dose group showed body descent within 30 minutes after dosing and then prostration. They died 40 minutes 4.5 hours after dosing after showing jumping and struggling, abnormal gait, and tremor.
Body weight:
Males from the 25, 50 and 200 mg/kg dose groups showed similar body weight changes as seen in the control group (treated orally with olive oil). Males dosed at 500 mg/kg showed restrained body weight increases on Day 2 and then a favorable increasing tendency thereafter. At the completion of the test, they were as heavy as the controI group.
Females dosed at 25, 50, and 200 mg/kg showed body weight changes similar to the change in the control group. Reduced body weight gain was seen in females from the 500 mg/kg dose group two days after dosing. Their body weights tended to increase thereafter; but were less than those of the control group animals throughout the test.
Gross pathology:
The decreased six males and six females from the 2000 mg/kg dose group showed mucosal bleeding in glandular stomach, intestinal tract, and anterior stomach, white anterior stomach mucous membrane, and intestinal tract mucosal colliquation.
No abnormalities were noted in males and females from the 25, 50, 200 and 500 mg/kg dose groups that survived the test period.

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LD50 of the test material was in the range between 500 and 2000 mg/kg.
Executive summary:

An acute oral toxicity test of the reaction product of 1,1'-(benzene-1,3 -diyl)bis(methylamine) and styrene was conducted in compliance with the OECD Test Guidelines No. 401 (1987) in the rat.

The reaction product was orally administered by gavage to SD rats (SPF). The tested concentrations lay in the range of 25 to 2000 mg/kg bw and 6 male and 6 female rats were treated per concentration. No abnormalities were detected in animals treated with 200 mg/kg bw or less. Animals treated with 500 mg/kg bw showed clinical signs (body descent, abnormal gait (male), squatting (female), diarrhea, loss of fur), but no abnormalities were observed on Day 3 or later. All animals treated with 2000 mg/kg bw died within 5 hours after application.

The LD50 of the test material was determined to be in the range of 500 and 2000 mg/kg. Accordingly, the reaction product falls into acute toxicity hazard category 4 according to Regulation (EC) No 1272/2008 (CLP).