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Description of key information

The results of these studies strongly indicate that the corrosive effects are the cause of all observed effects. Although a NOAEL was derived, it’s relevance is questionable. Only animals in the high-dose group showed significant effects, and these are consistent with the animals being dosed with a corrosive.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Remarks:
other: Read-across from sub-acute study
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-2013
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Animals were housed in controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 22+3°C, a relative humidity of 30-70% and 12-hour light/12-hour dark cycle.
Animals were housed in individually ventilated cages (IVC) - special animal room.
Polysulfone cages – floor area 900 cm2 containing sterilised clean shavings of soft wood were used.
Route of administration:
oral: gavage
Vehicle:
other: The test substance was administered suspended in aqua pro injectione.
Details on oral exposure:
The test substance was administered as suspension in aqua pro injectione using a stomach tube; oral application of rats was made daily.
Details on analytical verification of doses or concentrations:
The stability and the homogeneity of application form were determined in CETA analytical laboratories (Analytical Group I, Hana Teichmannová, Dipl. Eng). The results of analyses are stated in Annex 1 of the study report.
Duration of treatment / exposure:
28 days + recovery
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:
100mg/kg/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
200mg/kg/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
400mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
The study includes four main groups and two satellite groups of animals. Each main group consisted of 6 males and 6 females; each satellite group consisted of 6 males and 6 females. Main groups contained 3 treated groups of males and females (doses 100, 200, 400 mg/kg/day of body weight), and one control group (vehicle only). The satellite groups contained one control group (vehicle only), one treated group of males and females (400 mg/kg/day).
Control animals:
yes, concurrent vehicle
Details on study design:
The study includes four main groups and two satellite groups of animals. Each main group consisted of 6 males and 6 females; each satellite group consisted of 6 males and 6 females. Main groups contained 3 treated groups of males and females (doses 100, 200, 400 mg/kg/day of body weight), and one control group (vehicle only). The satellite groups contained one control group (vehicle only), one treated group of males and females (400 mg/kg/day).
The dose levels for males and females were chosen on the basis of results of Acute Oral Toxicity Study and DRFE (Dose-range finding experiment).
The administration period lasted 28 days. After that animals of main groups were sacrificed and satellite animals were observed for the next 14 days without treatment.
Positive control:
No
Observations and examinations performed and frequency:
Body weight: weekly
Food consumption: weekly
Water consumption: twice a week
Mortality control: daily
Health condition control: daily - during the acclimatization and the experimental part
General clinical observations: daily - during the administration period
Detailed clinical observations: once prior to start of administration, then in week intervals
Functional observations: in the last week of administration and recovery period
Laboratory examinations:
- urinalysis: 28th day of study (main groups)
and 42nd day of study (satellite groups)
- haematological examination: 29th day of study (main groups)
and 43rd day of study (satellite groups)
- biochemical examination: 29th day of study (main groups)
and 43rd day of study (satellite groups)
- biometry of organs: 29th day of study (main groups)
and 43rd day of study (satellite groups)
- pathological examination: 29th day of study (main groups)
and 43rd day of study (satellite groups)
- histopathological examination: from 29th day of study (main groups)
and from 43rd day of study (satellite groups)
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
effects observed, treatment-related
Behaviour (functional findings):
effects observed, treatment-related
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:
not specified
Details on results:
For males:
The test substance did not affect the body weight and body weight increment of males. Comparable values of treated and control animals were recorded during the application and recovery period.
Change of health condition of treated males from the dose level 400 mg/kg/day was recorded. Piloerection, irregular breathing, dyspnoea were noted in all animals of the dose level of 400 mg/kg/day. The same changes were observed in satellite treated males, but in recovery period (without treatment), only piloerection was recorded. The bad health condition of treated animals is considered to be treatment related.
During the haematological examination of males, statistically significant changes in haematology parameters were found in males of the dose level 100 mg/kg/day – the value of MCV was increased and in males of the dose level 200 mg/kg/day – the protrombin time was decreased. Also increased value of platelets count in all treated animals was recorded. Decreased values of total leucocytes count in males of the dose levels 200 and 400 mg/kg/day were noted. Changes were not treatment-related.
Biochemical examination showed statistically significant changes in males of the dose level 400 mg/kg/day– increased value of bilirubin and decreased value of ALP and chloride. Statistically significantly increased value of creatinine was recorded in males of the dose level 200 mg/kg/day. In satellite treated males, statistically significantly increased value of glucose and statistically significantly decreased value of bilirubin were reported.
The pH of urine was statistically significantly increased at the dose level 200 and 400 mg/kg/day. Also decreased urine volume at the dose level of 400 mg/kg/day was recorded with statistical significance.
Absolute and relative weights of organs of treated males were not affected by the test substance and were comparable to the control group. Only absolute weight of epididymides of satellite treated males was statistically significantly decreased (it is related to lower body weight of animals).
Macroscopical and microscopical findings were found only in one male of the satellite treated dose level, and this male had to be euthanasied prematurely by the reason of moribund condition. Extreme flatulence of stomach and intestine, dark foci on stomach and intestine were recorded at macroscopical examination. At histopathological examination, atrophy of thymus, vacuolar dystrophy of tubules of kidney, oedema of submucosis of forestomach, inflammatory infiltration and erosion of stomach, atrophy of spleen, hypertrophy of lymphatic tissue of caecum and small intestine and inflammatory lymphocyte infiltration of mucosa of large intestine were recorded. These findings are considered to be treatment related.


For females:
The body weight of females of the dose level of 400 mg/kg/day was decreased compared to control group during the whole application period. The necropsy body weight of females at this dose level was statistically significantly decreased. Decreased body weight was recorded also in satellite treated females, but at the end of recovery period, the body weight was comparable to control group. Decreased body weight of females at the dose level 400 mg/kg/day relates to decreased food consumption and is considered to be treatment related.
Change of health condition of treated females from the dose level 400 mg/kg/day was recorded. Piloerection, irregular breathing, dyspnoea were noted in all animals of this dose level. The same changes were observed in satellite treated females, but in recovery period (without treatment), only piloerection was recorded. The change of the health condition of females at the dose level of 400 mg/kg/day is considered to be treatment related.
During the haematological examination, decreased value of total leucocyte count and statistically significantly increased proportion of granulocytes and decreased proportion of lymphocytes in females of the dose level 400 mg/kg/day were recorded.
Statistically significantly increased value of bilirubin was recorded in females from the dose level 400 mg/kg/day at the biochemical examination. The value of ALP was statistically significantly decreased in females at dose level 100 mg/kg/day and the value of potassium was statistically significantly increased in females of the dose level 200 mg/kg/day. In satellite treated females, statistically significant changes in value of AST – decreased, and albumin – increased, were recorded.
The urine volume was decreased in treated females but without statistical significance and pH of urine was statistically significantly increased in females at the dose level 200 mg/kg/day/day.
The absolute weight of organs in treated group was well-balanced with the control group, except weight of liver in females of the dose level 400 mg/kg/day. Decreased weight of liver was recorded with statistical significance. Increased relative weight of kidneys, adrenal glands, ovaries and uterus in females at the dose level of 400 mg/kg/day were recorded. Statistically significantly increased relative weight of pituitary gland was recorded in treated satellite females.
Macroscopic findings were found in all females of the dose level 400 mg/kg/day. Flatulance of stomach and intestine were recorded in 3 females. Also erosion on stomach mucosa, decreased spleen, and decreased lymph nodes were recorded. These findings are treatment-related.
Microscopical findings were found in four females of the dose level 400 mg/kg/day. Histopathological findings in spleen – ischemia and atrophy, erosion and oedema of mucosa of forestomach, stomach - inflammatory infiltration and/or erosion, changes in intestine small and large and changes of lymph nodes were recorded. These finding are considered to be treatment related.

Changes in some of haematological and biochemical parameters in males and females of the dose levels 100 and 200 mg/kg/day were not connected with changes of organ structure and clinical changes of health condition of treated animals and therefore were not considered to be toxicologicaly important.

Negative effect of the test substance was mainly noticed in treated animals from the dose level 400mg/kg/day.
Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Critical effects observed:
not specified
Conclusions:
The results of these studies strongly indicate that the corrosive effects are the cause of all observed effects. Although a NOAEL was derived, it’s relevance is questionable. Only animals in the high-dose group showed significant effects, and these are consistent with the animals being dosed with a corrosive.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The results of this study strongly indicates that the corrosive effects are the cause of all observed effects. Although a NOAEL was derived, it’s relevance is questionable. Only animals in the high-dose group showed significant effects, and these are consistent with the animals being dosed with a corrosive.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Valid 28-day oral repeat-dse toxicity test in rat using the test substance.

Justification for classification or non-classification

The substabnce is severely irritating and corrosive to the skin, eyes and respiratory tract. Limited conclusions regarding systemic toxicity are possible due to the corrosive effects observed in animals (forestomach corrosion, low urinary pH, clinical indications).