Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From May 15th to July 3rd, 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Version / remarks:
July 31st, 1992
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Version / remarks:
May 12th, 1981
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Specific details on test material used for the study:
Stable in polyethylene glycol and water fo 48 hours.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Total number of animals: 30 males, 30 females
Total number of animals per group: Groups 1 and 4: 10 males, 10 females; Groups 2 and 3: 5 males and 5 females

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: PEG400 / bi-distilled water 1:1
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Duration of acclimatization period: 7 days
Duration of treatment: 28 days
Duration of recovery: 14 days
Frequency of treatment:
Dosing regime: 7 days/week
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Control
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Male: 10 animals at 0 mg/kg bw/day
Male: 5 animals at 50 mg/kg bw/day
Male: 5 animals at 200 mg/kg bw/day
Male: 10 animals at 1000 mg/kg bw/day

Female: 10 animals at 0 mg/Kg bw/day
Female: 5 animals at 50 mg/Kg bw/day
Female: 5 animals at 200 mg/Kg bw/day
Female: 10 animals at 1000 mg/Kg bw/day
Control animals:
yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:
Clinical signs, food consumption and body weights were recorded periodically during the treatment and recovery periods. Ophthalmoscopic examinations were performed at the end of the treatment ad recovery periods.
At the end of the dosing period and the treatment-free period blood samples were withdrawn for haematology and plasma chemistry analysis, and urine samples were collected for biochemical and microscopic analyses. All animals were killed, examine post-mortem and necropsied. Samples of major organs from all group 0 mg/kg and 1000 mg/kg animals and gross lesions from all animals were processed as haematoxylin and eosin stained slides and examine by light microscopy.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
There were no test substance-related clinical signs note in any group.
In one female treated with 100 mg/kg dose rales were noted on treatment days 11-12. No further clinical signs were observed in any other animal treated at 1000 mg/kg, therefore the rales were considered of no toxicological significance.
Description (incidence):
All animals survived the scheduled study period. One control female and one female treated with 1000 mg/kg dose died after blood sampling on the day of the scheduled necropsy after 4 weeks of treatment.
Body weight and weight changes:
no effects observed
Food efficiency:
no effects observed
Ophthalmological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Ophthalmologic findings were noted in small proportion of animals from all groups. They included persistent pupillary membrane, corneal opacity, anterior synechia and vitrous floaters. These findings occurred at similar incidences in the control and treated groups at the end of the treatment period. Therefore, they are considered to be unrelated to treatment with the test substance.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The only change of note was a slightly increate uric acid, total bilirubin, sodium, total protein and globulin level, and a slightly decreased chloride level in males treated with 1000 mg/kg at termination of the treatment. These findings, which were of minor degree and within historical control data, suggest metabolic adaptive changes rather than to constitute toxic effects. At the termination of the treatment-free recovery period these findings were generally reversed and found to be similar to those of the controls. All other statistical differences in the results of the haematology, clinical biochemistry and urinalysis data were considered to be incidental and unrelated to the treatment, and of normal biological variation for rats of this strain and age data for untreated Wistar rats.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
see haematological findings.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
see haematological findings.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Higher absolute and relative ovaries weights after 6 weeks were recorded in females at 1000 mg/kg. Both statistically significant findings were not considered to be test articles-related, but due to the slightly higher body weight of animals at 1000 mg/kg
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The macroscopic findings recorded were unremarkable and within the range of spontaneous alterations which may be seen in rats of this age and strain. They included dilated renal pelves, reddish discoloration in various organs and diluted uterine horns. Bluish discoloration of the gastrointestinal tract was noted amongst treated animals and was considered due to passive coloration by the test substance.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food efficiency
ophthalmological examination
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
Dose descriptor:
NOEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food efficiency
ophthalmological examination
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology

Target system / organ toxicity

Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
Based on the results of this study, 1000 mg/kg bw/day of the substance was established as the NOAEL and 200 mg/kg as the no-observed-effect-level NOEL.