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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:
From 10.01.1991 to 02.05.1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test method according to OECD guideline 407 and GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Camphen techn. rein
- Physical state: colorless, clear, vaseline-similar
- Analytical purity: 78.0%
- Purity test date: 28.09.1990
- Lot/batch No.: 54/90
- Stability under storage conditions: 12 months at room temperature
- Storage condition of test material: In the dark at room temperature
- Stability under test conditions: Guaranteed for four hours

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst AG, Box reason SPF breeding
- Age at study initiation: Approximately 6 weeks
- Housing: In air-conditioned spaces, in Makrolon cages (type 4) on granulate softwood, in groups of 5 animals.
- Diet (e.g. ad libitum): Rat diet Altromin 1324 (Altromin GmbH, Lage / Lippe), ad libitum, except the time in which the animals were in metabolism cages
- Water (e.g. ad libitum): Tap water in plastic bottles, ad libitum, except the time in which the animals were in metabolism cages
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 ºC
- Humidity (%): 50 ± 20 %
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 10.01.1991 To: 07.02.1991

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Sesame oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing solutions were prepared daily, immediately before the administration. The test material was homogeneously dispersed using a magnetic stirrer.
Volume administered: 5 mL/kg bw

VEHICLE
Oleum Sesami, Ph. Eur III, Fa Mainland, Pharmazeutische Fabrik GmbH, Ffm.
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
28 Days
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
62.5 mg/kg bw/day (1.25 % w/v)
Basis:
actual ingested
Remarks:
Doses / Concentrations:
250 mg/kg bw/day (5 % w/v)
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day (20 % w/v)
Basis:
actual ingested
No. of animals per sex per dose:
5 male and 5 female rats per group.
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for animal assignment (if not random): Randomization: Animals in cages randomisation = 946/90 and 947/90
Cages in the rack randomisation = 949/90

Examinations

Observations and examinations performed and frequency:
In all experimental groups, the behavior and general health of animals were observed twice daily during the experiment, once a day on weekends and holidays. Every week neurological disturbances, clouding of the ocular media, adverse effects on oral mucosa and tooth development disorders were studied.
The body weight was determined at the beginning of the experiment and twice a week.
Food consumption was determined twice a week and w ater consumption, once a week.
At the end of the experiment, the bllod count was investigated without prior food deprivation in all male and female animals. The blood was taken from the retro-orbital venous plexus. The following hematological parameters were determined: erythrocyte count, hemoglobin, hematocrit, MCV, MCH, MCHC, WBC, platelet count, differential count, reticulocyte count, Heinz'sche inner body, clotting time.
The urine tests were performed on all male and female animals and took place overnight from day 26 to 27 of the experimental period. The following parameters were determined: Appearance, color, pH, hemoglobin, protein, glucose, ketone body, bilirubin, urobilinogen, density and sediment.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Other examinations:
At the end of the study, animals were macroscopically examined. Alterations in organs were recorded, organs were weighed and their relative weights calculated. Histological preparations from the main organs were examined for microscopic changes. Chemical analysis were performed. The following parameters were analyzed: sodium, potassium, anorg. Phosphorus, uric acid, total and direct bilirubin, creatinine, serum glucose, urea nitrogen (BUN), calcium, chloride, AST (GOT), ALT (GPT), alkaline phosphatase (AP), gamma-glutamyl transferase (GGT), total protein, albumin.
Statistics:
Body weight, hematological and clinical tests, and organ weights (absolute and relative) were statistically compared with the control group.
The analysis were carried out using a program package for evaluation of toxicological tests, according to the Standard Operating Procedure (Department of Pharmaceutical Research).

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
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):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
The highest dose group (1000 mg/kg bw) showed an increased salivation. Behaviour and general health from other treated groups were not significantly different from control group. The body weight and food- and water-consumption were not affected by the administration of the test substance.
Haematological tests revealed no evidence of compound-related toxicity. In male animals from the highest dose group, clinical chemistry tests revealed an increase in urea nitrogen levels and a decrease in phosphorus levels. The urine was normal and showed no evidence of compound-related toxicity.
In male and female animals from the highest dose group, absolute and relative liver weights were increased.

Macroscopic examinations showed spotted kidneys in two male animals from the lowest dose group (62.5 mg / kg bw). In 3 mid-dose male animals and in all males from the highest dose group, colourless kidneys were observed. Animals from the highest dose group showed an increased vacuolization of the hepatocytes. Female animals from dose groups of 62.5 and 250 mg / kg bw did not show toxic effects. In all dose groups of male rats deposits of the test substance in the epithelium of the proximal renal tubules associated with necrosis of single cells have been observed. These effects seem to be specific for male rats and contingent upon alpha-2 microglobinemia. The renal toxic effects found in all dose levels groups in male rats are
interpreted as uniquely specific for male rats, and as having no relevance for other animal species and humans.

Based on the results of this study, the "'no observed effect level" (NOEL) for female rats was 250 mg / kg bw. For male rats, the NOEL could not be determined (it was lower than 63.5 mg/kg bw/day).

Effect levels

open allclose all
Dose descriptor:
NOEL
Effect level:
250 mg/kg bw/day (actual dose received)
Sex:
female
Basis for effect level:
other: overall effects
Dose descriptor:
NOEL
Effect level:
< 62.5 mg/kg bw/day (actual dose received)
Sex:
male
Basis for effect level:
other: renal toxic effects

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

For female rats the NOEL was 250 mg / kg bw/day. For male rats, the NOEL could not be determined (it was lower than 62.5 mg/kg bw/day).

Applicant's summary and conclusion

Conclusions:
For female rats the NOEL was 250 mg / kg bw/day. For male rats, the NOEL could not be determined (it was lower than 62.5 mg/kg bw/day).
Executive summary:

Camphene was daily administered to SPF Wistar rats (male and female) for 28 days at doses of 0, 62.5, 250 and 1000 mg / kg bw/day by gavage. Test method was according to OECD guideline 407. In all experimental groups, behaviour and general health were daily examined. The body weight and food consumption were determined twice a week, water consumption was determined once a week. Haematological and clinical tests, and urinalysis were also carried out. At the end of the study, animals were macroscopically examined. Alterations in organs were determined, organs were weighed and their relative weights calculated. Histologicalpreparations from the main organs were examined for microscopic changes. Body weight, haematological and clinical tests, and organ weights (absolute and relative) were statistically compared with the control group.

The highest dose group (1000 mg/kg bw/day) showed an increased salivation. Behaviour and general health from other treated groups were not significantly different from control group. The body weight and food- and water-consumption were not affected by the administration of the test substance.

Haematological tests revealed no evidence of compound-related toxicity. In male animals from the highest dose group, clinical chemistry tests revealed an increase in urea nitrogen levels and a decrease in phosphorus levels. The urine was normal and showed no evidence of compound-related toxicity.
In male and female animals from the highest dose group, absolute and relative liver weights were increased.

Macroscopic examinations showed spotted kidneys in two male animals from the lowest dose group (62.5 mg / kg bw/day). In 3 mid-dose male animals and in all males from the highest dose group, colourless kidneys were observed. Animals from the highest dose group showed an increased vacuolization of the hepatocytes. Female animals from dose groups of 62.5 and 250 mg / kg bw/day did not show toxic effects. In all dose groups of male rats deposits of the test substance in the epithelium of the proximal renal tubules associated with necrosis of single cells have been observed. These effects seem to be specific for male rats and contingent upon alpha-2 microglobinemia. The renal toxic effects found in all dose levels groups in male rats are interpreted as uniquely specific for male rats, and as having no relevance for other animal species and humans.

Based on the results of this study, for female rats the NOEL was 250 mg / kg bw/day. For male rats, the NOEL could not be determined (it was lower than 62.5 mg/kg bw/day).