Pyrrolo[3,4-c]pyrrole-1,4-dione, 2,5-dihydro-3,6-diphenyl-

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1987

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD Guideline Study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

May 12, 1981

GLP compliance:

yes

Remarks:

Research and Consulting Company AG, RCC Umweltchemie AG, Itingen, Switzerland

Limit test:

no

Test material

Test material information

Constituent 1

Details on test material:

- Physical state: solid
- Analytical purity: commercial grade
- Lot/batch No.: Op. 10002
- Stability under test conditions: stable for at least 2 hours
- Storage condition of test material: room temperature, dark

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: KFM Kleintierfarm Madoerin AG, Fuellinsdorf/Switzerland
- Age at study initiation: 7 - 9 weeks
- Weight at study initiation: males: 144 - 168 g, females: 151 - 179 g
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 40 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 400

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test article was weighed into a glass beaker on a tared Mettler PK 300 balance and the vehicle, polyethylene glycol (PEG 400) was added. The mixture was prepared daily prior to administration using a homogenizer and kept stable during application with a magnetic stirrer.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analyses are performed by photometric measurement.

Duration of treatment / exposure:

28 days

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300, 1000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based upon data received from the following acute studies carried out at RCC:
- acute oral toxicity in rats, RCC Project 062190, Ciba-Geigy Project 85 1229
- acute dermal toxicity in rats, RCC Project 062223, Ciba-Geigy Project 85 1232

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at termination of treatment
- Dose groups that were examined: all animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 4 weeks
- Anaesthetic used for blood collection: Yes: ether anesthesia
- Animals fasted: Yes
- How many animals: all
- Parameters examined: Erythrocyte count (RBC), Hemoglobin (HB), Hematocrit (HCT), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), Platelet count (PLATELETS), Reticulocyte count (RETIC), Nucleated erythrocytes normoblasts (NEN), Total leukocyte count (WBC), Differential leukocyte count (Diff. WBC Count), Red cell morphology, Thromboplastin time (PT), Partial thromboplastin time (PTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 4 weeks
- Animals fasted: Yes
- How many animals: all
- Parameters examined: Glucose, Urea, Creatinine, Bilirubin, total (BILL T), Cholesterol, total (CHOLEST. T.), Aspartate aminotransferase (ASAT(GOT)), Alanine aminotransferase (ALAT(GPT)), Lactate dehydrogenase (LDH), Alkaline phosphatase (ALP), Calcium, Phosphorus, Sodium, Potassium, Chloride, Albumin, Protein, total

URINALYSIS: Yes
- Time schedule for collection of urine: after 4 weeks
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters examined: Specific gravity (SPEC. GRAV.), pH, Protein, Glucose, Ketone, Bilirubin, Blood, Nitrite, Urobilinogen (UROBILI.), Urine Sediment (SED. MICRO.)

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

The following statistical methods were used to analyze the body weights, food consumption, organ weights and clinical laboratory data:
Univariate one-way analysis of variance was used to assess the significance of intergroup differences.
If the variables could be assumed to follow a normal distribution, the Dunnett-test (many to one t-test) based on a pooled variance estimate was applied for the comparison between the treated groups and the control groups.
The Steel-test (many-one rank test) was applied when the data could not be assumed to follow a normal distribution.
For the overall spontaneous mortality data, the Fisher's exact test for 2x2 tables was applied.
Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Individual values, means, standard deviations and statistics were rounded off before printing. For example, test statistics were calculated on the basis of exact values for means and pooled variances and then rounded off to two decimal places. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.

Results and discussion

Results of examinations

Details on results:

CLINICAL SIGNS AND MORTALITY: No death occurred. All animals survived to scheduled necropsy. No signs of toxicity which could be related to test article treatment were observed.

BODY WEIGHT AND WEIGHT GAIN: No significant differences in body weight gain was observed between the animals of the test article and control groups during the experiment.

FOOD CONSUMPTION: No significant differences in food consumption was observed between the animals of the test article and control groups during the experiment.

OPHTHALMOSCOPIC EXAMINATION: No treatment-related findings were noted.

HAEMATOLOGY: The assessment of hematology data indicated no changes of toxicological significance at termination of the treatment.

CLINICAL CHEMISTRY: The assessment of clinical biochemistry data indicated no changes of toxicological significance at termination of the treatment.

URINALYSIS: The assessment of urinanalysis data indicated no changes of toxicological significance at termination of the treatment.


ORGAN WEIGHTS: No significant differences were observed in absolute and relative organ weights between the animals of the control and test article-treated group.

GROSS PATHOLOGY: All pathologic findings recorded were of a spontaneous nature common to rats of this age and strain. There was no evidence of abnormal histopathological findings, resulting from treatment with the test substance.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

2

Dose descriptor:

NOEL

Effect level:

> 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Target system / organ toxicity

Critical effects observed:

not specified