Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
7 January 1988 - 4 February 1988
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report Date:
1988

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
Specific details on test material used for the study:
No further details specified on the study report.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
Specification: Sprague Dawley CFY rats obtained from Interfauna UK Limited, Wyton, Huntingdon, Cambridgeshire.
Justification: Preferred species of choice as historically used for safety evaluation studies and specified by appropriate regulatory authorities.
Sex:
male/female
Details on test animals and environmental conditions:
Specification: At the start of the main study the animals were aged 5 to 8 weeks. The weight variation did not exceed 20% of the mean weight.
Environment:
Temperature: 19 – 25 °C
Humidity: 45 – 75%
Lighting: 12 hours of artificial light in each 24-hour period
Ventilation: at least 15 air changes per hour.
Housing: Groups of five by sex in polypropylene cages with stainless steel lids suspended over trays containing absorbent paper.
Diet and Water: Rat and Mouse SQC. Expanded Diet No. 1 (Special Diet Services Limited, Witham, Essex, U.K.), and tap water ad libitum.
Acclimatisation period: Minimum 5 days.
Allocation: Animals will be allocated to dose groups using total randomisation procedure.
Identification: Each animal, selected at random, identified by ear-punch. Colour-coded cage card prepared with details of test material, project number, dose level, sex, numbers of animals, route of administration and Home Office licensee responsible for the study.

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
Gavage using a stomach tube attached to a graduated syringe.
Vehicle:
arachis oil
Details on oral exposure:
Preparation: The test material was dissolved in arachis oil B.P. weekly.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Analysis: No analysis of the test material formulations will be carried out.
Duration of treatment / exposure:
28 days
Frequency of treatment:
Once daily
Doses / concentrationsopen allclose all
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Four groups of 10 animals (5 male/5 female per group).
Control animals:
yes, concurrent vehicle
Details on study design:
Dose levels have been selected on the basis of the results from the 14-day Range Finding Test (Project Number: 10/207). Control animals received the vehicle alone in the amount received by the high dose group.
Positive control:
No.

Examinations

Observations and examinations performed and frequency:
Morbidity/Mortality Inspection: Twice daily during normal working week. Once daily at weekends.
Clinical Observations: Immediately before dosing and one hour after dosing. An additional observation was made five hours after dosing during the normal working week, not at weekends.
Bodyweights: Individual bodyweights are recorded on the day before the start of dosing and at weekly intervals.
Food Consumption: Diet intake recorded weekly for each cage group.
Laboratory Investigations: Haematological and blood chemical investigations were performed on all test and control group animals at termination (day 28). Blood samples were withdrawn under light ether anaesthesia from the orbital sinus.
Haematology;
Haemoglobin (Hb)
Haematocrit (HCT)
Red blood cell count (RBC)
Total white blood cell count (WBC)
Differential white blood cell count
Red blood cell indices
-mean cell haemoglobin (MCH)
-mean cell volume (MCV)
-mean cell haemoglobin concentration (MCHC)
Thrombotest Time (TT)

Blood Chemistry:
Blood urea Nitrogen
Total Protein
Albumin
Albumin/Globulin ratio (by calculation)
Sodium
Potassium
Chloride
Calcium
Inorganic Phosphorus
Creatinine
Total bilirubin
Alkaline phosphatase (AP)
Alkaline aminotransferase (ALAT)
Aspartate aminotransferase (ASAT)
Glucose
Sacrifice and pathology:
Post Mortem Studies: Carried out on all surviving animals (killed by intravenous sodium pentobarbitone) at termination and on animals that died or become moribund and killed during the study.
Gross Examination: Full external and internal examination of all animals.
Organ Weights;
Adrenals; Kidneys; Brain; Pituitary; Liver; Gonads; Spleen; Heart
Carried out on all survivors at termination.
Histopathology: Samples of the following tissues will be preserved from all animals in buffered formalin.
Adrenals Gross lesions
Aorta (thoracic) Muscle (skeletal)
Bone & Bone Marrow Pancreas
(sternum) Pituitary
Brain Rectum
Caecum Sciatic nerve
Colon Skin (hind limb)
Duodenum Spleen
Eyes Stomach
Heart Testes
Ileum Thymus
Jejunum Thyroid/parathyroid
Kidneys Trachea
Liver Urinary bladder
Lungs Oesophagus
Lymph nodes Ovaries

Initially the following tissues from control and high dose groups (Groups 1 and 4) will be examined microscopically.
Adrenals Liver
Gross lesions Spleen
Heart Target Organs
Kidneys
Other examinations:
No further examinations specified in the study report.
Statistics:
All data will be summarised in tabular form and analysed by appropriate statistical methods to assess the significance.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
No clinically observable signs of toxicity were noted during the study.
Mortality:
no mortality observed
Description (incidence):
There were no deaths.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Animals in all dose groups made expected bodyweight gains over the study period. Bodyweight gains in test animals were comparable to those seen in controls.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption in test animals was comparable with that seen in controls.
Food efficiency:
no effects observed
Description (incidence and severity):
Calculation of food efficiency showed no appreciable differences between groups.
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No intergroup differences were detected.
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
No treatment-related effects were demonstrated on any of the parameters measured.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No treatment-related effects were demonstrated on any of the parameters measured
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Several statisically significant differences were noted between organ weights for high dose males and those for control males.
Testes weights were reduced whilst heart and liver weights were increased. In all cases the differences were slight and there was no histopathological evidence of abnormalities in liver or heart.
All other organ weights, including kidney weights, were comparable between test and control animals.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Macroscopic examination revealed kidney abnormalities in two males and two females treated with 1000 mg/kg/day. The females showed pallor of the kidneys whilst the males showed enlargement of the right kidney. All kidneys appeared normal in intermediate and low dose group animals.
No further treatment-related macroscopic abnormalities were noted.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Treatment-related changes were observed in the kidneys of rats treated with 1000 mg/kg/day, but not at any other dose level.
Changes were characterised by basophilia and degeneration of the proximal tubular epithelium.
No further treatment-related changes were noted.
Histopathological findings: neoplastic:
not examined
Other effects:
no effects observed

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic

Target system / organ toxicity

Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
1 000 mg/kg bw/day (nominal)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
The NOAEL in the 28-d study (repeated dose, oral) for Dispersionsblau F 60 768 is 250 mg/kg bw in rats.
Executive summary:

The test material, Dispersionsblau F-60 768, was administered by gavage to three groups, each of five male and five female Sprague-Dawley (CFV) rats, for twenty-eight consecutive days, at dose levels of 50, 250 and 1000 mg/kg/day. A control group of five males and five females was dosed with vehicle alone (arachis oil B.P.).

 

Results

Mortality: There were no deaths.

 

Clinical Observations: No clinically observable signs of toxicity were noted during the study

 

Bodyweight: Animals in all dose groups made expected bodyweight gains over the study period.

Bodyweight gains in test animals were comparable to those seen in controls.

 

Food consumption: Food consumption in test animals was comparable with that seen in controls.

Calculation of food efficiency showed no appreciable differences between groups.

 

Water consumption: No intergroup differences were detected.

 

Haematology: No treatment-related effects were demonstrated on any of the parameters measured.

 

Blood chemistry: No treatment-related effects were demonstrated on any of the parameters measured.

 

Necropsy: Macroscopic examination revealed kidney abnormalities in two males and two females treated with 1000 mg/kg/day. The females showed pallor of the kidneys whilst the males showed enlargement of the right kidney. All kidneys appeared normal in intermediate and low dose group animals.

No further treatment-related macroscopic abnormalities were noted.

 

Organ weights: Several statistically significant differences were noted between organ weights for high dose males and those for control males.

Testes weights were reduced whilst heart and liver weights were increased. In all cases the differences were slight and there was no histopathological evidence of abnormalities in liver or heart.

All other organ weights, including kidney weights, were comparable between test and control animals.

 

Histopathology: Treatment-related changes were observed in the kidneys of rats treated with 1000 mg/kg/day, but not at any other dose level.

Changes were characterised by basophilia and degeneration of the proximal tubular epithelium.

No further treatment-related changes were noted.

 

Conclusion

The NOAEL for Dispersionsblau F-60 768 is 250 mg/kg.