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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG
CH 4414 Fuellinsdorf/Switzerland
- Age at study initiation: 8 weeks
- Weight at study initiation: males: 151 - 175 g; females: 152 - 179 g
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding ('Lignocel', Schill AG, 4132 Muttenz, Switzerland).
- Diet (e.g. ad libitum): Pelleted standard Kliba no. 343, Batches 35/88 and 36/88, rat maintenance diet
('Kliba', Klingentalmuehle AG, 4303 Kaiseraugst, Switzerland) ad libitum.
- Water (e.g. ad libitum):Community tap water from Itingen was available ad libitum.
- Acclimation period: Seven days under laboratory conditions, after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 40-70
- Air changes (per hr):10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/12 hours dark, music/light period.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilled water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test article was weighed into a glass beaker on a tared Mettler PE 360 balance and the vehicle, distilled water, 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:
not specified
Details on analytical verification of doses or concentrations:
Not available
Duration of treatment / exposure:
28 days
Frequency of treatment:
Once daily, 7 days per week
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 50, 200, 1000 mg/kg
Basis:
no data
No. of animals per sex per dose:
30 males
30 females
Control animals:
not specified
Details on study design:
Rationale:
Based upon data received from acute studies and the conclusions drawn from a 5-day oral toxicity (range-finding) study (RCC Project 223694). In this study, individuals of both test groups (200 and 1000 mg test article/kg bw) showed bluish to greenish discoloration of various organs, but no other treatment-related abnormalities occurred.
Positive control:
Not available

Examinations

Observations and examinations performed and frequency:
MORTALITY:
Observations for mortality were recorded once daily.

DETAILED CLINICAL OBSERVATIONS: Yes
Signs of toxicity were assessed once daily. Descriptions of all abnormalities were recorded and the subsequent progress was monitored

BODY WEIGHT: Yes
The body weight of each animal was recorded on the same days as the food consumption using the same recording system.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
The food consumption was recorded once during the acclimatization period and weekly thereafter using an on-line electronic recording system consisting of a
Mettler PK 4800 balance connected to the RCC computer.

OPHTHALMOSCOPIC EXAMINATION:
Ophthalmoscopic examinations were performed on all animals. A description of any abnormality was recorded. Examinations were performed at termination of
treatment and a second time on the recovery individuals of groups 1 and 4 at termination of the recovery period. Ten minutes after the application of a mydriatic solution (Dispersa AG, Winterthur / Switzerland) the cornea, lens, anterior chamber, vitreous body and ocular fundus of both eyes were examined under dimmed light using a Heine Miroflex 2 Ophthalmoscope (Eisenhut Vet. AG, Allschwil / Switzerland).

HAEMATOLOGY: Yes
The following anticoagulants were used during blood collection:
EDTA-K2 (hematology)
Sodium Citrate, 3.8% (coagulation, 1:10)
The following commercial reference controls were used to monitor the
performance of the method:
Hematology:
Eightcheck (normal range)
Eightcheck-L (lower abnormal range)
(TOA International Corporation, Kobe/Japan)
IL 282 CO-Oximeter Control
(Instrumentation Laboratory Inc., Lexington, Ma./USA)
Coagulation:
Ci-Trol-1 (normal range)
Ci-Trol-2 (high range)
(Merz & Dade AG, Duedingen/Switzerland)
19

Sacrifice and pathology:
ORGAN WEIGHTS
The following organ weights were taken from all animals necropsied at termination of treatment or recovery:
Liver; Kidneys; Adrenals; Testes.

NECROPSY AND HISTOPATHOLOGY
NECROPSY
Prior to necropsy, the animals were fasted for approx. 18 hours, but water was provided.
All animals were necropsied and descriptions of all macroscopic abnormalities were recorded. Necropsies were performed by experienced prosectors. All animals surviving to the end of the observation period were anesthetized by intraperitoneal injection of sodium pentobarbitone and killed by exsanguination.
after 4 weeks - February 7, 1989
after 6 weeks - February 21, 1989
Samples of the following tissues and organs were collected from all animals at necropsy and fixed in phosphate buffered neutral 4 % formaldehyde solution:
Adrenals; Aorta; Brain; Cecum; Colon; Duodenum; Epididymides; Esophagus; Eyes with optic nerve and Harderian gland; Female mammary gland area; Femur
including joint; Heart; Ileum; Jejunum; Kidneys; Larynx; Lacrimal gland, extraorbital; Liver; Lung infused with formalin; Lymph nodes, mandibular, mesenteric;
Nasopharynx; Ovaries; Pancreas; Pituitary gland; Prostate gland; Rectum; Salivary gland, mandibular, sublingual; Seminal vesicles; Sciatic nerve; Skeletal muscle; Skin; Spinal cord, cervical; Spleen; Sternum with marrow; Stomach; Testes; Thymus; Thyroid gland; Tongue; Trachea; Urinary bladder infused with formalin; Uterus with uterine cervix; Gross lesions.
HISTOTECHNIQUE / HISTOPATHOLOGY
The following tissues were trimmed, processed and embedded in paraffin wax and sectioned at a thickness of 2-4 micrometer: Adrenals, heart, kidneys, liver, spleen, lymph nodes, gastro-intestinal tract and gross lesions from all control and high dose animals, main test and recovery group.
Kidneys, liver, spleen lymph nodes, gastro-intestinal tract and gross lesions from the other treated groups. Sections were stained with haematoxylin and eosin and examined light microscopically.
Other examinations:
Not available
Statistics:
he 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 Dunnetttest (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 ophthalmoscopic data, the Fisher's exact test for 2 x 2 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 round-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

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
Administration of the test article did not cause any death throughout this study.
From day 9 of treatment onwards, the faeces of animals of all test articletreated groups were dark blue discolored. Test animals of group 4 recovered from this abnormality within 7 days of abstinence from the test article.

BODY WEIGHT AND WEIGHT GAIN
Changes of the body weight or body weight gain which could be related to the treatment with test article did not occur.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
The rates of food consumption were not affected significantly by the test article administered.

FOOD EFFICIENCY

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study)

OPHTHALMOSCOPIC EXAMINATION
No treatment-related changes were seen on ophthalmoscopy.

HAEMATOLOGY
The assessment of hematological, biochemical and urinalysis data indicated no changes of toxicological significance at termination of the treatment nor at the end of the treatment-free recovery period.

ORGAN WEIGHTS
Administration of test article did not affect significantly the organ weights and organ/body weight ratios during the treatment period. No treatment-related effects of toxicological relevance occurred

HISTOPATHOLOGY
Many treated animals were found to have green discoloration of the gastrointestinal tract and in lymphoid tissue. In the great majority of cases the discoloration was not apparent microscopically. The staining of tissues is thought to represent an inherent property of the compound and should not be regarded as an expression of toxicity. There were no other treatment related findings identified.

Effect levels

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
other: no adverse effects were seen

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The "no-observed-adverse-effect-level" of FAT 40'336/B is 1000 mg/kg bw for male and female rats when administered orally by gavage for a period of 28 days.
Executive summary:

In this subacute 28-day toxicity study, FAT 40'336/B was administered daily by gavage to SPF-bred Wistar rats. The study was comprised of four groups.

The dose levels used in this study are based on data from acute studies and the conclusions drawn from a 5-day oral toxicity (range-finding) study (RCC Project 223694). In this study, individuals of both test groups (200 and 1000 mg test article per kg body weight) showed bluish to greenish discoloration of various organs, but no other treatment-related abnormalities occurred.

Based upon the results obtained in this study, the "no-adverse-effect-level" of FAT 40'336/B is 1000 mg/kg body weight for male and female rats when administered orally by gavage for a period of 28 days. Oral administration of FAT 40'336/B to rats for 28 days at the dose level of 1000 mg/kg body weight/day produced no toxicological, hematological, biochemical or pathological evidence of toxicity.