Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27.3.-11.4.2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was carried out in accordance with internationally valid GLP principles.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

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:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeding farm BioTest s. r. o., 281 25 Konárovice, Czech Republic
- Age at study initiation: 7 - 9 weeks at the time application
- Weight at study initiation: range 150 - 170 g
- Fasting period before study: 20 h before study were not fed, water was given ad libitum
- Housing: animal room with monitoring conditions - 3 animals of one sex in one plastic breeding cage Velaz T4
- Bedding: sterilized shavings of soft wood
- Randomisation: according to the internal SOP 42, at the start of the study the weight variation of animals was minimal and did not exceed (+/-)
20% of the mean weight for each sex
- Identification of animals: colour marks 1-3 on tail of animals, each cage was marked with the number of study, sex and dose of the test substance
- Diet: ad libitum; ST 1 BERGMAN - standard pelleted diet ad libitum (producer: Mill Kocannda, Jesenice u Prahy, Czech Republic)
- Water: ad libitum; drinking tap water quality corresponding to Regulation No. 252/2004 Czech. Coll. of Law
- Acclimation period: 7 days
- Health condition: certificate of goodhealth condition, no signs of diseases were observed at clinical check-in during the acclimatisation period
and before the start of study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ( +/- )3; permanently monitored
- Humidity (%): 30 - 70 %; permanently monitored
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark


Study time schedule
Animal supply: 20.3. 2006
Experimental part of study: 27. 3. – 11. 4. 2006
Evaluation of results and final report elaboration: 12. 4. – 28. 4. 2006

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
Preparation and application of the test substance:
Immediately before application the test substance was weighed, mixed in vehicle (water for injections) and administered in a single dose by tube to
the stomach. The volume of administered suspension did not exceed 2 ml/100 g of body weight of animals.

CLASS METHOD
- Rationale for the selection of the starting dose: Test procedure with a starting dose of 2000 mg/kg was selected. The test substance in this dose level was administered sequentially to two groups of 3 females. The test substance administered at the dose of 2000 mg/kg caused death of two animals.

Testing schedule (according to EU Method B.1tris, Annex 1D)
START: 2000 mg/kg – 3 females (Step No.1): 1 female died ► 2000 mg/kg – 3 females (Step No. 2): 1 female died ► END of study
Test procedure with a starting dose of 2000 mg/kg was selected. The test substance in this dose level was administered sequentially to two groups of 3 females. The test substance administered at the dose of 2000 mg/kg caused death of two animals.


VEHICLE:
Aqua pro injectione (water for injections)
Biotika, a.s., Slovenská Lupča, SR
sterile, pellucid liquid without taste, colour and smell
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
6 (Step No.1-3 females, step No. 2-3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of weighing: before application, 8th and 15th day
- Frequency of observations: the first day: twice (30 minutes and 3 hours after application)
second and next days: twice per day (in the morning and in the afternoon) and daily thereafter for 14 days
- Necropsy of survivors performed: 15th day after application; by injection of veterinary preparation T 61 (1 ml i. v.). Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:
Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotoric activity, reaction to stimuli, presence of lacrimation, salivation anddichrage from nostrils, function of respiratory, digestive and urogenital system were recorded on special
data sheets. Results of clinical observations are presented in part Results and discussions.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
Step No. 1 : 1 female died
Step No. 2: 1 female died
Clinical signs:
Step No. 1 – 2000 mg/kg – 3 females
30 minutes after application: erected hair, hunched posture, anaemic cutis and anaemic mucous membrane and gasping (hard breathing) were
observed in all animals.
3 hours after application: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture and gasping (hard breathing) were
observed in all females. Females did not consume a feed.
2nd day after application died female No. 3 (20 hours after application). Erected hair, hunched posture, anaemic cutis and anaemic mucous
membrane were observed in two survivor animals. Females did not consume a feed.
3rd day after application: erected hair, hunched posture and anaemic cutis and anaemic mucous membrane were observed in two animals.
From 4th day after application no clinical signs of intoxication were detected in survivor females.

Step No. 2 - 2000 mg/kg – 3 females
30 minutes after application: erected hair, hunched posture, anaemic cutis and anaemic mucous membrane and gasping (hard breathing) were
observed in all animals.
3 hours after application: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture and gasping (hard breathing) were
observed in all females. Females did not consume a feed.
2nd day after application died female No. 2 (20 hours after application). Erected hair, hunched posture, anaemic cutis and anaemic mucous
membrane were observed in two animals. Females did not consume a feed.
From 3rd day after application no clinical signs of intoxication were detected in survivor females.
Details see in table No. 1 and 2
Body weight:
Average body weight in a group was counted from individual body weights.
Body weight increments were calculated from body weight at the start of the study and in the end of the study.
see table No. 3 and 4
The 8th day after application, the body weights of survivor animals were lower than at the beginning of the test. Total body weight increments were lower than it is used to be in the test animals of same age and sex.
Gross pathology:
All test animals that survived to the end of study were sacrificed on the 15th day by injection of veterinary preparation T 61 (1 ml iv.) and gross necropsy was carried out. Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated. All gross macroscopic changes of organs and tissues were recorded on special data sheets.
See table No. 5 and 6

Any other information on results incl. tables

Clinical observation

Table No. 1:  Clinical observation - 2000 mg/kg (Step No. 1)

Animal

No.

Death after

application

Observed changes

1

no

30 minutes: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping

3 hours: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping, no feed consuming

2nd day: erected hair, anaemic cutis and mucous membrane, hunched posture, no feed consuming

3rd day: erected hair, anaemic cutis and mucous membrane, hunched posture

4th – 14th day: no clinical signs of intoxication

2

no

30 minutes: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping

3 hours: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping, no feed consuming

2nd day: erected hair, anaemic cutis and mucous membrane, hunched posture, no feed consuming

3rd day: erected hair, anaemic cutis and mucous membrane, hunched posture

4th – 14th day: no clinical signs of intoxication

3

yes

30 minutes: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping

3 hours: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping, no feed consuming

2nd day: death

Table No. 2:  Clinical observation - 2000 mg/kg (Step No. 2)

Animal

No.

Death after

application

Observed changes

1

no

30 minutes: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping

3 hours: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping, no feed consuming

2nd day: erected hair, anaemic cutis and mucous membrane, hunched posture, no feed consuming

 3rd – 14th day: no clinical signs of intoxication

2

yes

30 minutes: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping

3 hours: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping, no feed consuming

2nd day: death

3

no

30 minutes: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping

3 hours: erected hair, anaemic cutis and anaemic mucous membrane, hunched posture, gasping, no feed consuming

2nd day: erected hair, anaemic cutis and mucous membrane, hunched posture, no feed consuming

 3rd – 14th day: no clinical signs of intoxication

 

Body weight

Table No. 3: Individual body weight (g) – 2000 mg/kg (Step No.1)

No.

 

Before application

8th day

15th day

Weight increment

1

174.58

197.56

216.60

42.02

2

166.24

164.45

198.15

31.91

3

161.21

-

-

-

Average

167.34

181.00

207.38

40.04

 

Table No. 4: Individual body weight (g) – 2000 mg/kg (Step No.2)

No.

 

Before application

8th day

15th day

Weight increment

1

182.88

175.66

215.94

33.06

2

172.11

-

-

-

3

163.34

148.85

186.69

23.35

Average

172.78

162.26

201.32

28.54

     

 Pathological examination 

 Table No. 5:  Pathological examination – 2000 mg/kg (Step No. 1) 

Animal No.

Necropsy findings

1

Liver - dystrophia

2

Liver - dystrophia

3

Stomach – dilatation, hyperaemia

Small intestine and stomach – flatulence

 

Table No. 6:  Pathological examination - 2000 mg/kg (Step No. 2) 

  Animal No.

Necropsy findings

1

Liver – dystrophia

Stomach – incrassate of surface in mucous part

2

Hypodermis– markedly yellow colour

Stomach – hyperaemia, acute catarrhal inflammation

Small intestine - acute catarrhal inflammation

3

Liver - dystrophia

Stomach– incrassate of surface in mucous part

 

Applicant's summary and conclusion

Conclusions:
The test substance toxicity was evaluated on the basis of mortality, clinical signs of intoxication, body weight increments during the observation period and necropsy findings in the end of study.
The test substance administered at the dose of 2000 mg/kg caused death of two animals (group No. 1 - first step:1 female died and group No. 2 - second step:1 female died), clinical signs of intoxication were observed in all six animals and macroscopic changes were diagnosed during pathological examination in all six animals.
Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance Potassium permanganate after a single oral administration to Wistar rats.

The testing was performed according to the Method B.1 tris Acute Oral Toxicity - Acute Toxic Class Method, Directive 2004/73/EC. Published in O.J. L152, 2004. 

Potassium permanganate was administered in a single dose as solution in water for injections, given orally via gavage to Wistar rats. 

The dosing was performed sequentially in two groups of three females (group No. 1 - first step and group No. 2 - second step) using the starting dose of 2000 mg/kg body weight. The volume did not exceed 2 ml/100 g body weight of animals.

 

The test substance administered at the dose of 2000 mg/kg caused death of two animals (group No. 1 - first step:1 female died and group No. 2 - second step:1 female died), clinical signs of intoxication were observed in both groups (all six animals) and macroscopic changes were diagnosed during pathological examination in all six animals. 

According to the study results, the value of LD50of the test substance Potassium permanganatefor female rats is higher than 2000 mg/kg of body weight.