Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
7.1.-8.9.2009
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
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): Potassium permanganate
- Molecular formula (if other than submission substance): KMnO4
- Molecular weight (if other than submission substance): 158.03
- Batch No.: 69
- Substance type: technical product
- Physical state: solid crystals
- Analytical purity: 99.42 % wt.
- Impurities (identity and concentrations): Manganese dioxide ca 0.1 % wt.
- Appearance: dark violet-purple crystalline powder with bronze lustre
- pH: 1% solution-6.1

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: SPF breeding, VELAZ s.r.o., Koleč u Kladna, Czech Republic, RČH CZ 21760152
- Age at study initiation: 6-7 weeks
- Fasting period before study: no
- Housing: Animals were housed in SPF animal room, 2-3 rats of the same sex (during acclimatization period) or individually (during treatment period) in one plastic cage (40x25x20cm) containing sterilised clean shavings of soft wood.
- Diet: ad libitum, Complete peleted diet for rats and mice in SPF breeding (ST 1 BERGMAN) was used, manufacturer: Ing. Miroslav Mrkvička – Výroba krmných směsí, Mlýn Kocanda, Kocanda No. 19, 252 42 Jesenice u Prahy. Diet was sterilised before using. Nutrient content of the diet: Crude protein – min. 21%, Drip – max. 14%, Fat – min. 3%, Fiber – max. 4.1%, Ash – max. 7%, Calcium – min. 1%, Phosphorus – min. 0.8%, Magnesium – min. 0.2%, Sodium – max. 0.25%. Composition of the diet: Wheat, Oats, Fish meal powder, Dried Snail-clover, Soya extracted groats, Wheat sprouts, Dehydrated yeast, Calcium carbonate, Vitamin and Mineral complex.
- Water: ad libitum, Free access to drinking water (water ad libitum). Water quality corresponded to Regulation No. 252/2004 Czech Coll. of Law, Health Ministry. Water was sterilised before using.
- Acclimation period: 5 days
- Additional Information: The standard pelleted laboratory animal diet is analysed for nutrients (once a year) and bacteriologically examined (every two months) on a regular basis. Results are retained in the CETA archives. Reports of analysis of water (performed twice a year) are retained in the CETA archives. Results of sterilizer effectivity control (performed once a year) are retained in the CETA archives. Analyses and controls mentioned above did not reveal any finding that could affect study integrity.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3°C
- Humidity (%): 30-70%
- Air changes (per hr): Animals were housed in controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle.


Study Time Schedule
Test substance delivery: 3. 3. 2006
Dose-range finding experiment: 7.1. – 6. 2. 2009
Main study
Date of animal arrival: 11. 2. 2009
Start of administration: 17. 2. 2009
End of administration: 19. 3. 2009
Clinical observation: 11. 2. - 19. 3. 2009 main groups
11. 2. - 31. 3. 2009 satellite groups
Urinalysis: 16. 3. - 19. 3. 2009 main groups
30. 3. - 31. 3. 2009 satellite groups
Blood taking and necropsies: 17. – 20. 3. 2009 main groups
31.3. –1. 4. 2009 satellite groups
Histopathological examination: 30. 6. – 21. 8. 2009
Evaluation of results and final report elaboration: 27. 7. – 8. 9. 2009

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Details on exposure:
TEST SITE
- % coverage: 10%
- Type of wrap if used: by foil and held in contact by non-irritating tape
- Time intervals for shavings or clipplings: Approximately 24 hours before testing, fur was shaved from dorsal area of the trunk of the test animals. No less than 10% of the body surface area was clear for the application of the test substance. Repeated shaving was needed at approximately weekly intervals.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test substance was removed using water
- Time after start of exposure:


TEST MATERIAL
- Amount(s) applied (volume or weight with unit):The concentrations of solutions in all dose levels were adjusted to ensure the administration of 2 mL per 100 g of body weight
- Constant volume or concentration used: yes
- The test substance was not possible to apply as solid substance because it is classified as corrosive and contact of crystallic form with skin causes skin marked irritation and full thickness destruction of skin tissue (see to the results of Acute Dermal Irritation/Corrosion Study No. 15/06/4; VUOS-CETA Report No. 0663; 2006).

VEHICLE
The test substance was administered dissolved in water for injectione.
Aqua for injections
Manufacturer: Ardeapharma, Ševětín, Czech Republic
Batch number: 0101210308, 0204190908

DOSE LEVELS AND GROUPS OF ANIMALS
- See table No. 1
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The determination of test substance was performed by a spectrophotometry.
Test substance stability and homogeneity were determined by measuring an absorbance of its water solution in visible range of spectrum.
Duration of treatment / exposure:
The treated and control groups were administered daily for the period of 28 days.
Frequency of treatment:
The animals were treated 7 days per week. The treatment starts each day at the same time (6.30 – 8.30 am).
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
150 mg/kg
Basis:
nominal per unit body weight
Remarks:
Doses / Concentrations:
300 mg/kg
Basis:
nominal per unit body weight
Remarks:
Doses / Concentrations:
600 mg/kg
Basis:
nominal per unit body weight
No. of animals per sex per dose:
20 males and 20 females - dose-range finding experiment
30 males and 30 females - main study
5 females+5 males per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
Before starting of main study the dose-range finding experiment with 14-day application period was performed. The dose levels 200, 500, 750 and 1000 mg/kg/day were derived from the limit dose 1000 mg/kg/day (according to the guideline). During dose-range finding experiment the body weight and clinical observations were recorded. Basic haematological examination and gross necropsy were made at the end of experiment.

Doses for the main study - 150, 300, 600 mg/kg/day were chosen on the basis of results of dose-range finding experiment.

- Rationale for animal assignment (if not random): Animals were randomly divided into the control and test groups and marked individually.
- Rationale for selecting satellite groups: random
- Post-exposure recovery period in satellite groups: 14 days after the end of application
- Section schedule rationale (if not random): all animals were sectioned
Positive control:
no

Examinations

Observations and examinations performed and frequency:
GENERAL CLINICAL OBSERVATION: Yes
- Time schedule: daily-during administration period
- All rats were observed daily during the administration period. This observation was made in order to record possible clinical effects after application and all changes in behaviour of animals. So it was done after application at the same time every day (c. from 13.30 to 15.30 p.m.) – at the time of expectation of maximal effect of the test substance. Animals were observed in natural conditions in their cages.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once prior to start of administration, then in week intervals
- This observation was carried out before the first application and then weekly. At the first part of observation behaviour of animals in the cage was monitored: posture, position of eyelids, tonic or clonic movements, piloerection, stereotypes or bizarre behaviour. The second part was the observation during removal from cage: reaction to handling, elasticity of skin, colour of mucous membranes, salivation, lacrimation, cleanliness of fur around foramina.

FUNCTIONAL OBSERVATION: Yes
- Time schedule: in the last week of study
- This observation was done at the end of administration period at main groups or at the end of recovery period at satellite groups. During functional examination, the sensory reactivity on auditory, visual, proprioceptive stimuli and pupillary reflex were evaluated and motor activity assessment was conducted. Moreover the individual observations of grip strength were performed using dynamometer. Measurements were made on: 1) pectoral legs, 2) pelvis legs, 3) all four legs. Grip power was expressed in Newtons.


HEALTH CONDITION CONTROL: Yes
- Time schedule: daily - during the acclimatization and the experimental part
- The health condition was controlled daily during the check-in, acclimatisation period, during the administration and during the recovery period in groups. Pre-experimental observation of all rats was performed to ensure that only the animals exhibiting normal behavioural activity would be entered into the study. In administration period this observation was performed before application.

MORTALITY CONTROL: Yes
- Time schedule: daily
- All rats were examined for vitality changes or mortality daily during the treatment and recovery periods.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly
- The body weight of animals was recorded on automatic balances with group average computing module. All animals were weighed immediately before euthanasia too. Weight increment was computed as an average per group per day (in grams).


FOOD CONSUMPTION: Yes
- Time schedule: weekly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- In the specified days every week the remainder of pellets was weighed in each cage, the new food was weighed out and the food consumption for the previous week was computed. Average values were calculated for each week of the study.


FOOD EFFICIENCY: Yes
- Food consumption for animal/day was calculated from average values of each group. Calculation of food conversion in %: weight increment/food consumption x 100.


WATER CONSUMPTION: Yes
- Time schedule for examinations: twice a week
- Drinking water consumption was recorded. Average values (water consumption per animal and per day) were calculated for each week of the study.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: 29th day of study (main groups) and 43rd day of study (satellite groups)
- Anaesthetic used for blood collection: Yes, light ether narcosis
- Animals fasted: No
- How many animals: all animals
- Parameters checked in table [No.2] were examined.


BIOCHEMICAL EXAMINATION: Yes
- Time schedule for collection of blood: 29th day of study (main groups) and 43rd day of study (satellite groups)
- Anaesthetic used for blood collection: Yes, light ether narcosis
- Animals fasted: No
- How many animals: all animals
- Parameters checked in table [No.3] were examined.


URINALYSIS: Yes
- Time schedule for collection of urine: 28th day of study (main groups) and 42nd day of study (satellite groups)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters checked in table [No.4] were examined.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- Time schedule: 29th day of study (main groups) and 43rd day of study (satellite groups)
During the necropsy a revision of the external surface of the body, of all orifices and the cranial, thoracic and abdominal cavities were carried out. Organs for consequent histopathological examination were taken out and stored in containers with fixative (buffered 4% formaldehyde).

HISTOPATHOLOGY: Yes
- Time schedule: 29th day of study (main groups) and 43rd day of study (satellite groups)
- Parameters checked in table [No.5] were examined.

BIOMETRY OF ORGANS: Yes
-Time schedule: 29th day of study (main groups) and 43rd day of study (satellite groups)
- At the end of study the experimental animals were narcotised and sacrificed by cutting the neck spine and medulla. After the gross necropsy of the cranial, thoracic and abdominal cavities the organs for weighing and further histological examination were collected. The absolute weights of liver, kidneys, adrenals, testes or ovaries, epididymides or uterus, thymus, spleen, brain, pituitary gland and heart were recorded. Afterwards the somatic indexes - SI (= relative weight of organ) were computed according to the following formula: SI = weight of organ x 100/ body weight.
Statistics:
The ANOVA test - Analysis of Variance (QC.Expert 2.5) at significance level 0.05 was used for the statistical analysis. This statistical analysis was used for the results of haematology, blood chemistry, urinalysis, biometry of organs and body weight. Control group with vehicle was compared with three treated groups and satellite control with vehicle was compared with satellite treated group.
The results statistically significant on probability level 0.05 are indicated by figures with asterisk in the tables of medians or averages.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Dermal irritation:
not examined
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
effects observed, treatment-related
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):
not examined
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 examined
Details on results:
CLINICAL SIGNS
- Males
The activity (poise, gait, reaction to handling) of all males at all treated groups was similar during the study and unchanging compared with activity of males at the control groups.
No changes were found out at all dose levels during the examinations of skin, hair, eyes, lacrimation, visible mucous membrane and respiration. Diarrhoeal was recorded in one male at the control satellite group. Excretion of other treated and control groups was without changes. Red secretion around eyes and nostrils were recorded in part of animals at all dose levels and at the control group.

- Females
The activity (poise, gait, reaction to handling) of all females at all treated groups was similar during the study and unchanging compared with activity of females at the control groups.
No changes were found out at all dose levels during the examinations of skin, hair, eyes, lacrimation, visible mucous membrane, excretion and respiration. Red secretion around eyes and nostrils were recorded in part of animals at all dose levels and at the control group.


MORTALITY
- Males
There were no unscheduled deaths during all the study.
- Females
There were no unscheduled deaths during all the study


BODY WEIGHT AND WEIGHT GAIN
1) AVERAGE BODY WEIGHT
- Males
During whole application period, the growth curve at all dose levels was slightly decreased in comparison to the control group but without statistical significance. Maximal difference was recorded at the highest dose level in the last week of treatment.
- Satellite males
The growth curve of treated males was relative well-balanced. Only slightly decreased in comparison to the control group (without statistical significance) was recorded in the last week of application (4th week of study).

- Females
During whole application period, the growth curves of the dose level 150 mg/kg/day was well-balanced with the control group. At the middle and highest dose levels slight decrease of body weight without statistical significance was recorded. Maximal difference was recorded at the middle dose level in the last week of treatment.
- Satellite females
The growth curve of treated females was well-balanced with the control group during whole application period. Slight increase of body weight of treated females was recorded in the last week of application.

2) BODY WEIGHT INCREMENT
- Males
After the 1st week of application the body weight increment of all treated groups was markedly decreased in comparison with the control group. At the end of the 2nd and 3rd weeks the body weight increments of all dose levels were higher than at the control group. At the end of application period the increment at all dose levels was markedly decrease again.
- Satellite males
The body weight increment of both groups was unbalanced during whole application. Markedly decreased increment of treated males was recorded in the first and last week of application period. On the contrary increased body weight increment was measured in the 2nd week of application and during the recovery period.

- Females
Decrease of body weight increment after the 1st week was recorded at the middle and highest dose levels. Slight increase of increment at the lowest and highest dose levels was measured in the end of the 2nd week. From the 3rd week to the end of application the increment of all dose levels was decreased again.
- Satellite females
The body weight increment of both groups was unbalanced during whole application. Increased increment of treated females was recorded in the 1st and 2nd week of application period. On the contrary increased body weight increment was measured in the 3rd week. From the 4th week of application and during the recovery period the body weight increment of both groups was relatively well-balanced.

FOOD CONSUMPTION
- Males
Food consumption of all treated groups was well-balanced with the control group during whole application. Slight decrease was recorded only at the highest dose level in the 1st week.
- Satellite males
The average food consumption of treated group was well-balanced with the control group during whole application and recovery period.

- Females
Food consumption of all treated groups was well-balanced with the control group during whole application.
- Satellite females
The average food consumption of treated group was well-balanced with the control group during whole application and recovery period.

FOOD CONVERSION
- Males
During the 1st week of application decreased food conversion was recorded at all dose levels. From the 2nd week to the 3rd of application the food conversion of the dose levels 150 and 300 mg/kg/day was well-balanced with the control group. At the highest dose level slightly increased food conversion was in the 3rd week. Markedly decreased conversation of all treated groups was measured at the last week of application.
- Satellite males
The food conversion of treated group was decreased in the first and last week of application. On the contrary increased conversion of treated group was recorded in the 2nd and 5th week of study. Relatively well-balanced food conversion of both groups was recorded only in the 3rd and the 6th week.
- Females
The well-balanced average food conversion of treated and control groups were recorded only at the dose level 150 mg/kg/day during the 1st and the 2nd week of study. Decreased conversion was recorded in the 2nd week of application at the dose levels 150 and during whole application period at the dose level 300 mg/kg/day. At the dose level 600 mg/kg/day decreased conversion was recorded at the 1st, 3rd and 4th week of study. Increase of food conversion was recorded only at the highest dose level in the 2nd week of application period.
- Satellite females
The average food conversion of both groups was unbalanced during application period. Increased conversion of treated group was recorded in the 1st and 2nd week of application and decreased conversion was in the 3rd week of study. In the last week of application period and during whole recovery period the food conversion of treated animals was well-balanced with the control group

WATER CONSUMPTION
- Males
The average water consumption of the treated groups was well-balanced with the control group only in the 1st week of application. Slightly increased consumption was recorded at the dose levels 150 mg/kg/day only in the 3rd week. At the dose levels 300 and 600 mg/kg/day slightly increase of water consumption was from 2nd week to the end of study.
- Satellite males

The water consumption of both groups was relatively well-balanced during whole application and recovery period.
- Females
The average water consumption of treated groups was relatively well-balanced with the control group in the first two weeks of application. Slightly increased consumption was recorded at the dose levels 150 and 600 mg/kg/day in the 3rd and 4th week. At the middle dose level the water consumption was well-balanced during whole study.
- Satellite females
In the treated group the water consumption was relatively well-balanced with the control group from the 3rd week of application to the end of recovery period. Increased consumption of treated animals was recorded in the first two weeks of application.



HAEMATOLOGY
- Males
Changes in differential leucocyte count were recorded. Value of monocytes was increased with dependence on the dose at the all dose levels. Statistical significance was found out at the middle and highest dose levels. Accompanied decreased value of lymphocytes without statistical significance was also recorded at these dose levels. Total leucocyte count was slightly increased (without statistical significance) at the dose level 150 mg/kg/day. Prolonged protrobine time was measured at the lowest and middle dose levels but statistical sinficance was recorded only at the dose level 300 mg/kg/day. Prolonged APTT (without statistical significance) was measured at the dose levels 300 and 600 mg/kg/day.
Other measured parameters were similar to the control group.
All observed parameters were within historical control range.
- Satellite males
Statistically significantly increased value of platelet count was recorded at the treated group.
Other measured parameters were similar to the control group.

- Females
Changes in differential leucocyte count were recorded. Value of monocytes was statistically significantly increased with dependence on the dose level at the all dose levels. Accompanied decreased value of lymphocytes with statistical significance was recorded at the highest dose level. Total leucocyte count was slightly increased (without statistical significance) at the dose level 150 mg/kg/day. Prolonged protrobine time was measured at the highest dose levels but without statistical significance. Prolonged APTT (without statistical significance) was measured at the dose levels 150 and 600 mg/kg/day.
Other measured parameters were similar to the control group. Statistical analysis of the data revealed no significant intergroup differences.

- Satellite females
No statistically significant changes were found out.
All measured parameters were similar to the control group.

BIOCHEMICAL EXAMINATION
- Males
All measured parameters at the dose level 150 mg/kg/day were similar to the control group. Statistical analysis of the data revealed significant intergroup differences at the dose levels 300 and 600 mg/kg/day. The sodium concentration was significantly increased in both dose levels. Significantly increased activity of ALT was recorded only at the dose level 300 mg/kg/day. Slightly increased activity of AST without statistical significance was measured also at the middle treated group. Insignificant decreased concentration of glucose was found out at the middle and highest dose levels. Slightly decreased value of creatinine and concentration of potassium were recorded at the highest dose level. Concentration of chlorides was insignificantly decreased at the lowest dose level.
- Satellite males
Increased concentration of phosphorus and decreased concentration of potassium and chloride with statistical significance was recorded at treated group. Slightly increased value of urea and decreased activity of AST were recorded at treated group without statistical significance.
All other measured parameters were similar to the control group.

- Females
Statistical analysis of the data revealed no significant intergroup differences. Slightly increased activity of AST at the highest dose level and ALT at the middle dose level were recorded. Decreased value of creatinine without statistical significance was measured at the middle and highest dose levels. All other measured parameters were similar to the control group
- Satellite females
Statistically significantly increased value of glucose and albumin was recorded at treated group. Concentration of chloride and sodium ions was significantly decreased also at this dose level. Insignificantly increased concentration of bilirubine and activity of AST was measured at the treated group. All other measured parameters were similar to the control group.


URINALYSIS
- Males
No statistically significant changes in urine were recorded at any dose levels. Slight decrease of urine volume was recorded at the dose level 600 mg/kg/day. Blood was detected in two males and nitrites in one male at the dose level 150 mg/kg/day. Urine of one animal at the middle dose level showed white cloud. Presence of leucocytes was recorded in one male at the highest dose level.
- Satellite males
Statistical analysis of the data revealed significant intergroup differences. Significantly decreased urine volume and increased pH of urine were recorded at the treated group. Urine of one treated male showed blood in urine. Also presence of leucocytes was recorded in one treated male.

- Females
There were no treatment-related changes in urinary parameters.
Statistical analysis of the data revealed no significant intergroup differences.
- Satellite females
There were no treatment-related changes in urinary parameters.
Statistical analysis of the data revealed no significant intergroup differences.


ORGAN WEIGHTS
Absolute Organ Weight
- Males
Statistical analysis of the data revealed no significant intergroup differences.
Treated animals showed slightly decreased weight of liver with dependence on the dose level. The weight of other organs was well-balanced at all groups.
- Satellite males
Absolute weight of all organs was well-balanced at both groups. Statistical analysis of the data revealed no significant intergroup differences.

- Females
Animals at the dose level 300 mg/kg/day showed statistically significantly decreased of weight of kidneys. Slightly decreased weight of kidneys was recorded also at the lowest and highest dose levels but without statistical significance. The weight of liver was slightly decreased at the middle dose level (without statistical significance). Slightly decreased absolute weight of pituitary gland and ovaries was found out at the highest dose level. Other organs was relatively well-balanced.
- Satellite females
Absolute weight of organs was well-balanced at both groups. Statistical analysis of the data revealed no significant intergroup differences.

Relative Organ Weight
- Males
Relative weight of organs was well-balanced at all dose levels. Statistical analysis of the data revealed no significant intergroup differences.
- Satellite males
Slightly insignificantly increased weight of liver was recorded at the treated group. Relative weight of other organs was well-balanced with the control group. Statistical analysis of the data revealed no significant intergroup differences.

- Females
Animals at the dose level 150 mg/kg/day showed slightly decreased weight of liver and kidneys (without statistical significance). Relative weight of other organs was relatively well-balanced. Statistical analysis of the data revealed no significant intergroup differences.
- Satellite females
The weight of liver in treated group was statistically significantly increased. Relative weight of other organs was relatively well-balanced at both groups.

PATHOLOGY
Macroscopic Findings
- Males
During the macroscopic examination no important pathologic changes were found. In 2-1-0-0 males no macroscopic changes were observed.
Examination of application area revealed changes on skin and regional lymph nodes: brown colouring of skin in 0-0-5-0 males, punctiform plagues in 0-0-2-0 males, dry and red foci of skin in 0-0-0-5, congested lymph nodes in 3-4-5-4 males and enlarged lymph nodes in 0-0-2-0 males.
In thoracic cavity focal changes (changed colour or petechial foci) in lungs were diagnosed in 0-0-0-2 males and petechie on thymus was in 0-0-0-1 male.
In abdominal cavity the irregular colour of kidneys were only found in 1-0-0-1 males.
In cranial cavity no changes were diagnosed.
- Satellite males
During the macroscopic examination of males no important pathologic changes were found. In 5-2 males no macroscopic changes were observed.
Examination of application area showed congested or enlarged regional lymph nodes only in 0-1 male.
In thoracic cavity only focal changes in lungs in 0-1 male were recorded.
In abdominal cavity no changes were recorded.In cranial cavity no changes were diagnosed.

- Females
During the macroscopic examination of females no important pathologic changes were found. In 3-0-0-0 females no macroscopic changes were observed.
Examination of application area revealed changes on skin and regional lymph nodes: brown colouring of skin in 0-0-5-0 females, punctiform plagues in 0-0-2-0 females, dry and red foci of skin in 0-0-0-5, crust on skin in 0-0-0-1 female, congested lymph nodes in 2-4-5-3 females and enlarged lymph nodes in 0-0-2-0 females.
In thoracic cavity only petechie on thymus in 0-1-0-0 female were found out.
In abdominal cavity irregular colour of liver in 0-0-2-0 females and marked structure in 0-0-1-0 female were observed. Dilatation of uterus with fluidwas diagnosed in 3-2-3-1 females.
In cranial cavity no changes were diagnosed.
- Satellite females
During the macroscopic examination of females no important pathologic changes were found. In 3-3 females no macroscopic changes were observed.
Examination of application area showed no changes.
In thoracic cavity only petechie on thymus in 0-1 female and focal changes in lungs (grey focus) in 1-0 female were recorded.
In abdominal cavity only irregular colour of liver in 0-1 female was recorded. Dilatation of uterus with fluid was diagnosed in 1-2 females.
In cranial cavity no changes were diagnosed.


HISTOPATHOLOGY: NON-NEOPLASTIC
- Males
Incidence of pathological affection of skin application area was increased with dependence on the dose level. Focal inflammation of skin in 0-2-3-5 males and parakeratosis or hyperkeratosis in 0-0-1-3 males were recorded. Application area without changes was found out in 5-3-1-0 males. Occurence of findings in regional lymph nodes was well-balanced at all treated and control groups: erythrocytosis in 2-4-5-4 males and hyperplasia in 5-4-5-5 males. Haemorrhage in thymus was recorded in 2-2-0-1 males.
Incidence of pathological findings in digestive system was sporadic. Atrophy of phundal glands in stomach in 2-1-2-1 males, inflammation in liver in 1-2-3-2 males and proliferation of oval cells in liver of 0-1-0-1 males were recorded. In genital tract oedema of interstitium in prostate gland of 2-5-3-0 males, inflammation of prostate gland in 0-2-2-0 males and inflammation of epididymis in 1-1-2-1 males were found out. Dystrophy in cortex of kidneys was recorded in 4-3-2-0 males. Cartilagenous metaplasia in heart was recorded in 2-0-0-1 males. Other microscopical findings were recorded only sporadically (affected only one or two males in whole study).
- Satellite males
Application area was without changes. Occurence of findings in regional lymph nodes was well-balanced at treated and control groups: erythrocytosis in 2-3 males, plasmocytosis 1-0 male and hyperplasia in 3-2 males. Atrophy of phundal glands in stomach of 1-1 males, inflammation in liver of 4-5 and foci of oval cells in liver of 0-3 males were found out in digestive system. Dystrophy in cotrtex of kidneys was recorded in 2-1 males. In genital tract oedema of interstitium in prostate gland of 4-1 males, inflammation of prostate gland in 0-2 males and inflammation of epididymis in 1-2 males were found out. Other microscopical findings were recorded only sporadically (affected only one male).

- Females
Incidence of pathological affection of skin application area was recorded only in treated groups. Focal inflammation of skin in 0-0-2-5 females, crusts on epidermis in 0-0-2-0 females and parakeratosis in 0-0-0-3 females were recorded. Application area without changes was found out in 5-5-3-0 females. Occurence of findings in regional lymph nodes was relative well-balanced at all treated and control groups: erythrocytosis in 4-4-5-5 females and hyperplasia in 5-5-5-5 females.
In digestive tract the higher incidence of pathological affections was recorded in liver. Inflammation in 5-4-4-4 females, pigmentation in 0-1-0-2 females and proliferation in 2-2-1-2 females were found out. Focal infiltration in oesophagus was found out in 1-2-0-0 females. Cartilagenous metaplasia in heart was recorded in 0-1-1-1 females. Inflammation or haemorrhages in lungs were found out in 2-2-0-0 females. In genital tract only hydrometra of uterus was recorded in 3-2-2-1 females. Other microscopical findings were recorded only sporadically (affected only one or two females in whole study).
- Satellite females
Application area was without changes. Pathologic changes were found out in digestive system. Hyperplasia in 5-2 females and erythrocytosis in 1-3 females were recorded in regional lymph nodes. In digestive system foci of oval cells in liver of 4-2 females and inflammation in liver of 5-4 females were found out. Cartilagenous metaplasia in heart was recorded in 2-1 females. Hydronephrosis in kidneys was recorded in 1-1 females Pigmentation in spleen was recorded in 2-2 females. In genital tract only hydrometra of uterus was recorded in 1-2 females. Other microscopical findings were recorded only sporadically (affected only one female).

Effect levels

Dose descriptor:
NOAEL
Effect level:
150 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
haematology
clinical biochemistry
urinalysis

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The dermal administration of the test substance Potassium permanganate to rats for a period of twenty-eight consecutive days at dose levels 150 mg/kg/day produced no toxicologically significant changes in the parameters measured. No major functional changes in any organ systems or severe organ dysfunction were detected. Histopathological examination revealed no pathological changes, which could be related with administration of the test substance at the dose level 150 mg/kg/day. Significant changes in clinical biochemistry, haematology and urinalysis parameters, which indicate organism dysfunction, were recorded at the dose level 300 and 600 mg/kg/day.

Based on the results of laboratory investigations in clinical biochemistry, haematology and urinalysis and with respect to the results of histopathological examination the following conclusion about NOAEL can be suggested in this study:
Executive summary:

Methods

   SPF Wistar rats were used for testing. The test substance was administered dissolved in water for injections. The six groups of animals were included in the study – 4 main groups and 2 satellite groups.

   Main groups contained one control group (vehicle only) and three treated groups (doses 150, 300 and 600 mg/kg/day). Satellite groups contained one control group (vehicle only) and one treated group (600 mg/kg/day). The administration of test and control substances lasted 28 days. After that the satellite animals were observed for next 14 days without treatment.

   The stability and the homogeneity of the application form in water for injections were determined in CETA analytical laboratories. The concentrations of suspensions in all three dose levels were adjusted to ensure the administered volume of 2 mL per 100 g of body weight and over an area, which was approximately 10% of the total body surface area. 

   Before starting of main study the dose-range finding experiment with 14-day application period was performed. The dose levels 200, 500, 750 and 1000 mg/kg/day were derived from the limit dose 1000 mg/kg/day (according to the guideline). During dose-range finding experiment the body weight and clinical observations were recorded. Basic haematological examination and gross necropsy were made at the end of experiment.

 Doses for the main study - 150, 300, 600 mg/kg/day were chosen on the basis of results of dose-range finding experiment. 

   During the 28-day study clinical observation and health status control were performed daily. The body weight, food consumption were measured weekly and the detailed clinical observation was carried out in the same time interval. Water consumption was measured twice a week. Before the end of study the functional observation was accomplished. The study was finished by urinalysis, haematological and biochemical analysis, and gross necropsy of animals. The selected organs for weighing and histopathology examination were removed.

  

Results

   There were no unscheduled deaths during the test. At the middle and highest dose levels the application of the test substance caused reversible changes of application area with histopathological findings (focal inflammation of skin, erythrocytosis and hyperplasia of regional lymph nodes) which faded away after recovery period (without application).

   The animal’s health condition was very good during whole study. Clinical signs of toxicity were detected sporadically at all treated and control group. Functional observation evidenced no effect of the test substance.

   Slight effect on body weight gain was detected in both sexes which was more marked in males at the middle and highest dose levels. The body weight and body weight increment of test groups was statistically insignificantly decreased during whole application period. No adverse effect on dietary intake was detected. Unbalanced (mainly decreased) food conversion was recorded at all treated group males and females but markedly at the middle and highest dose levels. The water consumption was slightly unbalanced during whole study.

    Changes of differential leucocyte count were recorded in both sexes with statistical significance at the end of application. Increased total leucocyte count was also measured at the end of application but without statistical significance. Thehaemocoagulationexamination detected prolonged PT and APTT in both sexes but these changes achieved the statistical significance only in males at the middle dose level. These changes were not recorded in satellite treated males after 14-day recovery period.

    Statistical analysis of biochemicalparametersrevealed significant differences in males (increased sodium concentration against control at the middle and highest dose levels). Significant changes of ions concentration were recorded in both sexes at the end of recovery period.Due to presence of changed urinary parameters (decreased urine volume and increased pH of urine) this was considered to be toxicological important. Other intergroup differences considered to be adaptative response to stress without toxicological significance.

   Pathology examination revealed some but not treatment related changes in digestive system (atrophy of phundal glans in stomach, foci of oval cells or inflammation in liver), pigmentation in spleen, dystrophy in cortex of kidneys and cartilagenous metaplasia in heart. The intergroup differences were minimal, therefore they were regarded as fortuitous. Further sporadic pathologic changes were recorded in reproductive system: inflammation and oedema of interstitium in prostate gland, inflammation in epididymis, in males and hydrometra of uterus and pigmentation in spleen in females. Frequencies of these microscopic findings were similar in treated and control groups. This was considered to be of no toxicological importance. No neoplastic findings were recorded by histopathological examination.