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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28.3.-28.4.2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Test performed before release of OECD 429

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
- Substance type: technical product
- Physical state: solid
- Analytical purity: 99.42
- Impurities (identity and concentrations): Manganese dioxide cca 0.1 %
- Lot/batch No.: 69

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Albino guinea pigs, BFA
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeding farm Bio Test Konárovice, 281 25 Czech Republic
- Age at receiving: 6-7 weeeks
- Weight at study initiation: Exposed group 406,42(+/-)53,04
Control group: 379,87(+/-)50,95
- Housing: Monitored conditions, microbiologically defined background, according to SOP No. 40
- Diet: Pelleted standard diet for experimental animals. ad libitum, MAK 1, made by Bergman, Kocanda Mill
- Water: Drinking tap water, ad libitum, with ascorbic acid, quality corresponding to Regulation No. 252/2004 Czech Coll. of Law.
- Acclimation period: 6 days
- Bedding: Sterilized shavings of soft wood
- Random selection: According to SOP No.42
- Identification of animals: Individual labelling of cages and labelling of the animals
- Stock and health condition: No signs of disease were observed at clinical check-in
- Prophylactic arrangement: Cleaning and disinfection of animal room was performed regularly as it is described in appropriate SOP No.10

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3 °C, permanently monitored
- Humidity (%): 30-70 %, permanently monitored
- Air changes (per hr): approximately 15 air changes
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle: 6 am-6 pm/6 pm-6 am


IN-LIFE DATES: From: 27.3.2006 To: 11.4.2006

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: water for injections and vaselinum album
Concentration / amount:
Induction - intradermal injections: 1 % test substance in water for injections
Induction - topical application: 10 % test substance in vaseline
Challenge- topical application: 0.1 % test substance in vaseline.

Total amount of the test substance used for main test was 2.135g:
Induction - intradermal injections – 0.12g
Induction - topical application - 2g
Challenge – 0.015g.
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: water for injections and vaselinum album
Concentration / amount:
Induction - intradermal injections: 1 % test substance in water for injections
Induction - topical application: 10 % test substance in vaseline
Challenge- topical application: 0.1 % test substance in vaseline.

Total amount of the test substance used for main test was 2.135g:
Induction - intradermal injections – 0.12g
Induction - topical application - 2g
Challenge – 0.015g.
No. of animals per dose:
35 animals
5 for pilot experiment
30 for main study
20 animals in treatment group (6 males and 14 females)-teratment group
10 animals in control group (6 males and 4 females)-control group
Details on study design:
- Clinical signs: daily, clinical symptoms of intoxication and health condition of animals were observed.
- Mortality/viability: daily
- Body weight: 0 and 24 day, the body weight of animals was recorded one day before the first application (day 0) and after observation of skin
reaction (day 24). The average of group was counted from individual body weight values of animals.


- The skin reaction: 24, 48 and 72 hours after intradermal injection
48, 72 and 96 hours after induction – topical application



MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: --
- Test groups: TS in FCA, TS in water(for injection), TS in vaseline (for topical)
- Control group: FCA, water
- Site: shoulder region
- Frequency of applications: 1 st day -intradermal injections and 7th day-topical application
- Duration: 0-9 d
- Concentrations: 1%, 10%


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 20
- Exposure period: 24 h
- Test groups: TS in vaseline
- Control group: TS in vaseline
- Site: L flank
- Concentrations: 0,1%
- Evaluation (hr after challenge): 48, 72 h
Positive control substance(s):
yes
Remarks:
Benzocaine (ethyl p-aminobenzoate)

Results and discussion

Positive control results:
Reliability of the experimental technique is checked periodically in about six-month interval by the experiment with known sensitiser benzocaine. For intradermal application the 5% concentration in olive oil was used and the challenge exposure was performed by the same concentration in vaselina alba. The result of last experiment (March 2006): positive skin reaction – 4 animals (total number of animals = 10) i.e. 40%. According to the guideline the minimal number of animals with positive skin reaction is expected to be 30%. The result is satisfactory.
The results of reference study test are described in VUOS-CETA internal report No. 0643.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no pathological reaction of skin
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.1 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no pathological reaction of skin.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
0.1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no pathological reaction of skin
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 0.1 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no pathological reaction of skin.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.1%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no pathological reaction of skin
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0.1%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no pathological reaction of skin.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
0.1%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no pathological reaction of skin
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 0.1%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no pathological reaction of skin.
Reading:
other: periodical check of the system
Group:
positive control
Dose level:
5% in olive oil
No. with + reactions:
4
Total no. in group:
10

Any other information on results incl. tables

Traditional sensitisation test

Body weight

The body weight of animals was recorded one day before the first application (day 0) and after observation of skin reaction (day 24). The average of group was counted from individual body weight values of animals.

Table 1.: Body weight in grams– mean values +SD

 

Number of animals

Start of study

End of study

Increment

Exposed group

20

406.42±53.04

539.14±68.62

+132.72

Control group

10

379.87±50.95

507.33±87.46

+127.46

Evaluation of skin reaction after induction

Intradermal injections

The skin reaction evaluation of the test area of all animals from the exposed group showed moderate erythema around the sites of intradermal injections puncture. These changes faded away after 48 hours.

The skin reaction evaluation of the test area of all animals from control group showed discrete erythema around first sites of intradermal injections puncture. These changes faded away after 24 hours.

 

Topical application The evaluation of skin reactions of exposed group showed in all animals a moderate confluent erythema without oedema in the test area with fade-out time 72 hours.The evaluation of skin reactions in control group with vaseline did not show any changes in skin of all animals.

 

Evaluation of skin reactions after challenge

Exposed group at 48 hours after the challenge

The evaluation of skin reactions in the exposed group carried out at 48 hoursafter the start of the challenge phase of study did not show any changes on skin of exposed left flank of animals. The other flank of the animals applied with the vaseline only, did not show any skin reactions in all the 20 animals.

 

Exposed group at 72 hours after the challenge

The evaluation of skin reactions in the exposed group carried out at72 hours after the start of challenge phase of study did not show any changes in skin of the exposed flank of all animals. The other flank did not show any changes in skin of all the 20 animals.

 

Control group at 48 and 72 hours after the challenge

   The evaluation of skin reactions in the control groupcarried out at 48 and 72 hours after the start of the challenge phase of study did not show any changes in skin of animals in both flanks.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Conclusions:
The exposed animals showed no pathological skin reactions and no other negative clinical symptoms throughout the experiment.
The body weight of animals increased throughout the study and it was not affected by the treatment. In the course of experiment the animals did not show any clinical symptoms of intoxication.
The test substance, Potassium permanganate, was not shown to be a contact allergen in guinea pigs.
Executive summary:

The test substance,Potassium permanganate,was tested for the assessment of skin allergic effects using albino guinea pigs (strain BFA).

 

      The test was performed according to the OECD Test Guideline No. 406, Skin sensitisation, which is analogous to the EU Method B.6, Skin sensitisation.

 

    The Magnusson and Kligman maximization procedure was followed. The test was performed on albino guinea pigs (12 males and 23 females).

    The experiment proceeded in three phases: two induction phases - topical application and the challenge phase. Potential skin reactions were evaluated at the end of experiment.

    The exposed animals showed no allergic skin reactions and no other negative clinical symptoms throughout the experiment.

    

     The test substance, Potassium permanganate, was not shown to be a contact allergen in guinea pigs.