Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19-jun-2012 to 10-jul-2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Details on test material:
- Name of test material (as cited in study report): 4,4’-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with butan-1-ol and 3-aminomethyl-3,5,5-trimethylcyclohexylamine
- Substance type: Clear colourless viscous liquid
- Physical state: viscous liquid
- Purity: 100% (UVCB)
- Lot/batch No.: WA 522
- Expiration date of the lot/batch: 01 January 2014
- Storage condition of test material: At room temperature in the dark

Test animals

Species:
rat
Strain:
other: Wistar strain, Crl:WI (Han)
Sex:
female
Details on test animals or test system and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 9-10 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean (first group: 150-162 grams; second group: 165-172 grams; third group: 161 - 182 grams).
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
-Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

IN-LIFE DATES: From: 19 June 2012 to 10 July 2012

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (10 mL/kg) body weight.

DOSAGE PREPARATION:The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for density of the test substance and 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. No correction for purity was made.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Based on the test substance properties and in the interest of animal welfare, a dose level of 300 mg/kg bw was selected for the first group to minimize any testing likely to produce severe responses in animals.
Doses:
300 mg/kg body weight
2000 mg/kg body weight

No. of animals per sex per dose:
2000 mg/kg: 3 females
300 mg/kg: 6 (2 groups of three females in a stepwise manner)
Control animals:
no
Details on study design:
The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. Based on the test substance properties and in the interest of animal welfare, a dose level of 300 mg/kg bw was selected for the first group to minimize any testing likely to produce severe responses in animals. 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. As no death occurred in the 300 mg/kg bw treated animals, three further females were treated at a dose level of 2000 mg/kg bw. As all 2000 mg/kg bw treated animals died within 2 days after treatment a further group of three female animals was treated at a dose level of 300 mg/kg bw..

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/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 (if found dead after Day 1).
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: The animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
- Other examinations performed: none.
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
All animals at 2000 mg/kg were found death on Day 1 or 2. No mortality occurred at 300 mg/kg.
Clinical signs:
Clinical signs observed during the study period were as follows:
Dose level Clinical signs
2000 mg/kg All animals showed lethargy, hunched posture, uncoordinated movements, slow breathing, piloerection, ptosis and/or hypothermia up until death on Day 1 or 2.
300 mg/kg Hunched posture and piloerection were noted in all animals on Day 1 and/or 2. One animal showed also lethargy, tremor of the head and uncoordinated movements on Day 1.
Body weight:
The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:
Dark foci in the gastro-intestinal tract, dark red discoloration and irregular surface of the stomach, and dark red discoloration of the thymus were found in the animals that died during the study, at macroscopic post mortem examination. Macroscopic post mortem examination of the surviving animals at 300 mg/kg at termination did not reveal any abnormalities.

Applicant's summary and conclusion

Conclusions:
In an acute oral toxicity study with female rats, performed according to OECD/EC test guidelines, an LD50 300-2000 mg/kg bw was determined.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 1000 mg/kg body weight.