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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:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
May-July 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: 1a: Guideline study (GLP)
Justification for type of information:
Read across to an analogue based on structural similarity. An analogue justification is attached to section 13 of the dataset.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
read-across source
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May-July 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: 1a: Guideline study (GLP)
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Version / remarks:
(July 1995)
Deviations:
yes
Remarks:
Protein and creatinine excretion was not measured (as an error). These measurements performed in the study T8073730 (OECD 421) were also reported in this dossier.
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Animals were supplied by 'Harlan GmbH', Borchen, Germany and were about 4 weeks old. Males had an initial weight of 97 (83-114)g and females of 101 (95-108).
Housing in groups of 2 or 3 rats in Makrolon(R) cages Type IV on wood granules. All animals or the study were kept in the same room with no other animals from other studies and in an 20 Pa overpressure environment at 22°C, 55 +/-5% relative humidity and a 12 hour light/dark cycle and low-noise music.
Diet was standard diet "Provimi Kliva 3883.0.15" pellets supplied by Provimi Kliva SA, Kaiseraugst, Switzerland ad libitum. Water was provided ad libitum.
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
vehicle: demineralized water
administration volume: 10 ml/kg bw
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
not applicable
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg/body weight
Basis:
actual ingested
No. of animals per sex per dose:
5 males + 5 females
Control animals:
yes, concurrent vehicle
Details on study design:
5 male and 5 female Wistar rats/dose and control group
ADMINISTRATION/EXPOSURE:
vehicle: demineralized water
administration volume: 10 ml/kg bw
Positive control:
not applicable
Observations and examinations performed and frequency:
CLINICAL OBSERVATION AND FREQUENCY:

Inspection of Animals:
for morbidity and mortality: Twice daily, once daily on weekends and public holidays
general clinical examinations (in cage): Daily
detailed clinical examinations: Weekly
open field observation (OFO): Once before start and once weekly thereafter
Ophthalmology: At study begin: All rats.
At termination: Control and high dose
Functional Observational Battery: Day 19-20 (relative)
Motor Activity: Day 18-19 (absolute)
Determination of body weight(s): Daily
food consumption: Weekly
water consumption: Weekly

CLINICAL PATHOLOGY at the end of study:

HEMATOLOGY:
Leukocytes, Erythrocytes, Hemoglobin, Hematocrit, Reticulocytes, Differential blood count, Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), HQUICK, Platelets

CLINICAL CHEMISTRY:
Alanine aminotransferase, Alkaline phosphatase,Aspartate aminotransferase,Gamma glutamyl transferase, Glutamate dehydrogenase, Lactate dehydrogenase, Creatin kinase, Albumin, Cholesterol, Creatinine, Glucose, Total protein, Urea, Potassium, Sodium, Calcium, Chloride, Inorganic Phosphate

URINALYSIS
- Quantitatively: Density and Volume (Protein and creatinine excretion was not measured as an error.
- Semi-quantitatively: pH, Glucose, Blood, Bilirubin, Ketone bodies, Urobilinogen, Protein
- Microscopy of sediment
Sacrifice and pathology:
ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC):

- Gross Pathology:
Fixed organs: Adrenals, aorta, brain (cerebrum, cerebellum, ponslmedulla), epididymides, esophagus, eyes, eyelids, extraorbital lachrymal glands, femur, harderian glands, head (with nasal and paranasal cavities), heart, intestine (duodenum, jejunum, ileum, cecum, colon, rectum and remaining intestine), kidneys, larynx, liver, lymph nodes (mandibular, bronchial/hilus, and mesenteric), lung, mamma, optical nerves, ovaries, oviducts, pancreas, pharynx, pituitary, prostate, salivary glands, sciatic nerve, seminal vesicles (incl. coagulating glands), skeletal muscle (thigh), skin (mammary and muzzle), spinal cord (cervical, thoracic, lumbar), spleen, sternum, stomach (forestomach and glandular stomach), testes, thymus, thyroids (including parathyroid glands), tongue, trachea, ureter, urethra, urinary bladder, uterus with uterine cervix, vagina, Zymbal’s glands and all organs or tissues with macroscopic findings.

- Microscopic: Adrenals, aorta, brain (cerebrum, cerebellum, ponslmedulla), epididymides, esophagus, femur, heart, intestine (duodenum, jejunum, ileum, cecum, colon, rectum and remaining intestine), kidneys, liver, lung, lymph nodes, ovaries, oviducts, pancreas, pituitary, prostate, salivary glands, sciatic nerve, seminal vesicles (incl. coagulating glands), skeletal muscle (thigh), skin (mammary and muzzle), spinal cord (cervical, thoracic, lumbar), spleen, sternum, stomach (forestomach and glandular stomach), testes, thymus, thyroids (including parathyroid glands), trachea, urinary bladder, uterus with uterine cervix, and all organs or tissues with macroscopic findings. Slides were prepared from the first five animals of all groups and evaluated from the control and the high concentration group.


ORGAN Weights:
Adrenals, brain, epididymides, heart, kidneys, Liver, Ovaries, Spleen, Testes, Thymus, uterus
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
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Details on results:
The water intake was slightly increased in 1000 mg/kg males. There was no correlate to this finding and a toxicological relevance is excluded.
Hematology: at 1000 mg/kg MCH was slightly increased in females (p<0.01) and MCHC was sligthly increased in both sexes (p<0.05).
Clinical chemistry: females exhibited a decreased mean LDH activity.
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: not applicable
Dose descriptor:
NOEL
Effect level:
ca. 300 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: increased mean corpuscular hemoglobin in females and increased mean corpuscular hemoglobin concentration in both sexes at 1000 mg/kg bw.
Critical effects observed:
not specified

NOAEL: 1000 mg/kg b.w./d (nominal);

NOEL 300 mg/kg b.w./d (nominal) increased MCH in females and increased MCHC in both sexes.

MORTALITY:

No animal died up to and including 1000 mg/kg bw.

CLINICAL SIGNS

No clinical signs were observed up to and including 1000 mg/kg bw.

BODY WEIGHTS:

No statistically significant and dose-depending changes in body weights or body weight gain were evident up to and including 1000 mg/kg bw in males and females. As result of blood and urin sampling body weights were reduced from day 25 to day 27.

FOOD CONSUMPTION:

No toxicologically relevant changes in food intake were observed up to and including 1000 mg/kg in males and females.

WATER CONSUMPTION:

No toxicologically relevant changes in water intake were observed up to 300 mg/kg in males and 1000 mg/kg in females. At 1000 mg/kg a slight increase (13%) of water intake per kg body weight was seen in males.

HEMATOLOGY and CLINICAL PATHOLOGY:

Hematological parameters were not influenced by the test substance up to 300 mg/kg. At 1000 mg/kg slightly elevated MCH (females, p=0.01) and MCHC (both sexes) were measured (p=0.05).

The parameters of clinical chemistry were unchanged up to 300 mg/kg. At 1000 mg/kg females exhibited a decreased mean for the LDH activity. Bearing in mind that, generally, an increased but not decreased LDH activity reflects a toxic effect, this finding is of no toxicological relevance.

None of the urine parameters was treatment-related changed up to and including 1000 mg/kg.

OPHTHALMOLOGY:

No treatment-related changes at 1000 mg/kg

FUNCTIONAL OBSERVATIONAL BATTERY (FOB)/MOTOR ACTIVITY (MA/LMA):

FOB: No statistically significantly and dose-dependently differing up to and including 1000 mg/kg.

MA/LMA: No indication of test substance-related changes up to and including 1000 mg/kg.

GROSS PATHOLOGY:

No macroscopical organ changes attributable to the test substance were detected up to and including 1000 mg/kg.

ORGAN WEIGHTS:

None of the absolute and relative organ weights were statistically significantly and dose-dependently changed up to and including 1000 mg/kg.

HISTOPATHOLOGICAL FINDINGS:

An increased hemopoiesis in the spleen by severity (mean severities with ascending dose: 1.6-2.0-2.0-2.6) was detected in females at 1000 mg/kg, which is interpreted as an adaptive response to the treatment. Indications on an anemia were not noted.

In the adrenal glands vacuolation of the zona fasciculata was raised (incidence with ascending dose: 1-1-0-4) in males at 1000 mg/kg. Due to their localization in the upper adrenal cortex the vacuoles may probably derive from accumulations with cholesterol and/or glucocorticosteroids (lipidosis).

Reason / purpose for cross-reference:
read-across: supporting information

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Version / remarks:
(July 1995)
Deviations:
yes
Remarks:
Protein and creatinine excretion was not measured (as an error). These measurements performed in the study T8073730 (OECD 421) were also reported in this dossier.
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Voranol RA 800; Molecular Mass: (average Mn = 280 g/mol)
; Units of Propylene oxide, approx: 70-90%, Units of ethylene oxide: approx 10-30%
Lot #: UB06083011
Colourless viscous Fluid

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Animals were supplied by 'Harlan GmbH', Borchen, Germany and were about 4 weeks old. Males had an initial weight of 97 (83-114)g and females of 101 (95-108).
Housing in groups of 2 or 3 rats in Makrolon(R) cages Type IV on wood granules. All animals or the study were kept in the same room with no other animals from other studies and in an 20 Pa overpressure environment at 22°C, 55 +/-5% relative humidity and a 12 hour light/dark cycle and low-noise music.
Diet was standard diet "Provimi Kliva 3883.0.15" pellets supplied by Provimi Kliva SA, Kaiseraugst, Switzerland ad libitum. Water was provided ad libitum.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
vehicle: demineralized water
administration volume: 10 ml/kg bw
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
not applicable
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg/body weight
Basis:
actual ingested
No. of animals per sex per dose:
5 males + 5 females
Control animals:
yes, concurrent vehicle
Details on study design:
5 male and 5 female Wistar rats/dose and control group
ADMINISTRATION/EXPOSURE:
vehicle: demineralized water
administration volume: 10 ml/kg bw
Positive control:
not applicable

Examinations

Observations and examinations performed and frequency:
CLINICAL OBSERVATION AND FREQUENCY:

Inspection of Animals:
for morbidity and mortality: Twice daily, once daily on weekends and public holidays
general clinical examinations (in cage): Daily
detailed clinical examinations: Weekly
open field observation (OFO): Once before start and once weekly thereafter
Ophthalmology: At study begin: All rats.
At termination: Control and high dose
Functional Observational Battery: Day 19-20 (relative)
Motor Activity: Day 18-19 (absolute)
Determination of body weight(s): Daily
food consumption: Weekly
water consumption: Weekly

CLINICAL PATHOLOGY at the end of study:

HEMATOLOGY:
Leukocytes, Erythrocytes, Hemoglobin, Hematocrit, Reticulocytes, Differential blood count, Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), HQUICK, Platelets

CLINICAL CHEMISTRY:
Alanine aminotransferase, Alkaline phosphatase,Aspartate aminotransferase,Gamma glutamyl transferase, Glutamate dehydrogenase, Lactate dehydrogenase, Creatin kinase, Albumin, Cholesterol, Creatinine, Glucose, Total protein, Urea, Potassium, Sodium, Calcium, Chloride, Inorganic Phosphate

URINALYSIS
- Quantitatively: Density and Volume (Protein and creatinine excretion was not measured as an error.
- Semi-quantitatively: pH, Glucose, Blood, Bilirubin, Ketone bodies, Urobilinogen, Protein
- Microscopy of sediment
Sacrifice and pathology:
ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC):

- Gross Pathology:
Fixed organs: Adrenals, aorta, brain (cerebrum, cerebellum, ponslmedulla), epididymides, esophagus, eyes, eyelids, extraorbital lachrymal glands, femur, harderian glands, head (with nasal and paranasal cavities), heart, intestine (duodenum, jejunum, ileum, cecum, colon, rectum and remaining intestine), kidneys, larynx, liver, lymph nodes (mandibular, bronchial/hilus, and mesenteric), lung, mamma, optical nerves, ovaries, oviducts, pancreas, pharynx, pituitary, prostate, salivary glands, sciatic nerve, seminal vesicles (incl. coagulating glands), skeletal muscle (thigh), skin (mammary and muzzle), spinal cord (cervical, thoracic, lumbar), spleen, sternum, stomach (forestomach and glandular stomach), testes, thymus, thyroids (including parathyroid glands), tongue, trachea, ureter, urethra, urinary bladder, uterus with uterine cervix, vagina, Zymbal’s glands and all organs or tissues with macroscopic findings.

- Microscopic: Adrenals, aorta, brain (cerebrum, cerebellum, ponslmedulla), epididymides, esophagus, femur, heart, intestine (duodenum, jejunum, ileum, cecum, colon, rectum and remaining intestine), kidneys, liver, lung, lymph nodes, ovaries, oviducts, pancreas, pituitary, prostate, salivary glands, sciatic nerve, seminal vesicles (incl. coagulating glands), skeletal muscle (thigh), skin (mammary and muzzle), spinal cord (cervical, thoracic, lumbar), spleen, sternum, stomach (forestomach and glandular stomach), testes, thymus, thyroids (including parathyroid glands), trachea, urinary bladder, uterus with uterine cervix, and all organs or tissues with macroscopic findings. Slides were prepared from the first five animals of all groups and evaluated from the control and the high concentration group.


ORGAN Weights:
Adrenals, brain, epididymides, heart, kidneys, Liver, Ovaries, Spleen, Testes, Thymus, uterus

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
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Details on results:
The water intake was slightly increased in 1000 mg/kg males. There was no correlate to this finding and a toxicological relevance is excluded.
Hematology: at 1000 mg/kg MCH was slightly increased in females (p<0.01) and MCHC was sligthly increased in both sexes (p<0.05).
Clinical chemistry: females exhibited a decreased mean LDH activity.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: not applicable
Dose descriptor:
NOEL
Effect level:
ca. 300 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: increased mean corpuscular hemoglobin in females and increased mean corpuscular hemoglobin concentration in both sexes at 1000 mg/kg bw.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

NOAEL: 1000 mg/kg b.w./d (nominal);

NOEL 300 mg/kg b.w./d (nominal) increased MCH in females and increased MCHC in both sexes.

MORTALITY:

No animal died up to and including 1000 mg/kg bw.

CLINICAL SIGNS

No clinical signs were observed up to and including 1000 mg/kg bw.

BODY WEIGHTS:

No statistically significant and dose-depending changes in body weights or body weight gain were evident up to and including 1000 mg/kg bw in males and females. As result of blood and urin sampling body weights were reduced from day 25 to day 27.

FOOD CONSUMPTION:

No toxicologically relevant changes in food intake were observed up to and including 1000 mg/kg in males and females.

WATER CONSUMPTION:

No toxicologically relevant changes in water intake were observed up to 300 mg/kg in males and 1000 mg/kg in females. At 1000 mg/kg a slight increase (13%) of water intake per kg body weight was seen in males.

HEMATOLOGY and CLINICAL PATHOLOGY:

Hematological parameters were not influenced by the test substance up to 300 mg/kg. At 1000 mg/kg slightly elevated MCH (females, p=0.01) and MCHC (both sexes) were measured (p=0.05).

The parameters of clinical chemistry were unchanged up to 300 mg/kg. At 1000 mg/kg females exhibited a decreased mean for the LDH activity. Bearing in mind that, generally, an increased but not decreased LDH activity reflects a toxic effect, this finding is of no toxicological relevance.

None of the urine parameters was treatment-related changed up to and including 1000 mg/kg.

OPHTHALMOLOGY:

No treatment-related changes at 1000 mg/kg

FUNCTIONAL OBSERVATIONAL BATTERY (FOB)/MOTOR ACTIVITY (MA/LMA):

FOB: No statistically significantly and dose-dependently differing up to and including 1000 mg/kg.

MA/LMA: No indication of test substance-related changes up to and including 1000 mg/kg.

GROSS PATHOLOGY:

No macroscopical organ changes attributable to the test substance were detected up to and including 1000 mg/kg.

ORGAN WEIGHTS:

None of the absolute and relative organ weights were statistically significantly and dose-dependently changed up to and including 1000 mg/kg.

HISTOPATHOLOGICAL FINDINGS:

An increased hemopoiesis in the spleen by severity (mean severities with ascending dose: 1.6-2.0-2.0-2.6) was detected in females at 1000 mg/kg, which is interpreted as an adaptive response to the treatment. Indications on an anemia were not noted.

In the adrenal glands vacuolation of the zona fasciculata was raised (incidence with ascending dose: 1-1-0-4) in males at 1000 mg/kg. Due to their localization in the upper adrenal cortex the vacuoles may probably derive from accumulations with cholesterol and/or glucocorticosteroids (lipidosis).

Applicant's summary and conclusion