Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
28.03.2006 - 28.04.2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study performed according to the guideline.

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.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Details on test material:
The appearance of Potassium permanganate is dark violet - purple crystalline powder with bronze lustre.

Potassium permanganate has been used as a surrogate for sodium permanganate where data are not available. Read-across from potassium permanganate to sodium permanganate is appropriate from the toxicological point of view as the most toxicologically relevant part of the substances is the same (permanganate). The contribution of the sodium/potassium ions to the toxicity of the respective substances is likely to be minimal. The toxicity of both substances is therefore likely to be very similar and will be dominated by local (site of contact) irritant/corrosive effects and systemic toxicity due to the absorption of manganese ions. This toxicophore similarity is adequate justification for waiving the conduct of specific studies with sodium permanganate and the dossier reflects this waiving proposal by including summaries of read-across studies where appropriate.

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: BFA
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
The test was performed on albino guinea pigs, 12 young adult males and 23 females, nulliparous and non-pregnant (5 animals for the pilot, 30 for the main study) . The source of the animals was a breeding farm BioTest Konarovice, 281 25 Czech Republic. Age on arrival was 6-7 weeks, and animals were acclimitised for 6 days. Drinking water was supplied unrestricted, pelleted standard diet (MAK1) was fed ad libitumand animals were housed individually. No signs of disease were observed at clinical check-in.
ENVIRONMENTAL CONDITIONS
The room temperature (22±3oC) and relative humidity (30-70%) was continously monitored with 12 hour light/dark cycle.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
other: two induction phases: intradermal followed by topical
Vehicle:
other: vaselinum album (Batch number 416/02) for topical applicaions and and water for injection (Batch number 3130605) for intradermal injections
Concentration / amount:
The appropriate concentrations of the test substance determined from the pilot experiment were as follows: Induction - intradermal injections: 1% test substance in water for injection, Induction - topical application: 10% test substance in vaseline and Challenge - topical application. Total amount of test substance used for the main test was 2.135g. Induction - intradermal injection = 0.12g, Induction - topical application = 2g and Challenge = 0.015g.
Challengeopen allclose all
Route:
other: topical application
Vehicle:
other: vaselinum album (Batch number 416/02) for topical applicaions and and water for injection (Batch number 3130605) for intradermal injections
Concentration / amount:
The appropriate concentrations of the test substance determined from the pilot experiment were as follows: Induction - intradermal injections: 1% test substance in water for injection, Induction - topical application: 10% test substance in vaseline and Challenge - topical application. Total amount of test substance used for the main test was 2.135g. Induction - intradermal injection = 0.12g, Induction - topical application = 2g and Challenge = 0.015g.
No. of animals per dose:
5 females were used in the pilot experiment, 6 males and 14 females in the main test and 6 males and 4 females in the control group.
Details on study design:
The Magnusson and Kligman maximization procedure was followed. The experiment proceeded in three phases: two induction phases - topical application and the challenge phase.

Induction: Intradermal injections
Day 0 - treated group
Three pairs of intradermal injections of 0.1 mL volume were given in the shoulder region, cleared of hair so that one of each pair lies on either side of the midline.
Injection 1: a 1:1 mixture (v/v) FCA/water for injection
Injection 2: the test substance 1% in water for injection
Injection 3: the test substance 1% in water for injection in 1:1 mixture (v/v) FCA/water for injection

Day 0 - control group
Three pairs of intradermal injections of 0.1 mL volume were given in the same sites as in the treated animals.
Injection 1: a 1:1 mixture (v/v) FCA/water for injection
Injection 2: water for injection
Injection 3: a 1:1 mixture (v/v) FCA:water for injection


Induction: Topical application
Day 6 - treated group
The test area was cleared of hair. A filter paper (2 x 4cm) saturated with 10% test substance in vaseline was applied to the test area and held in contact by an occlusive dressing for 48 hours.
Day 6 - control group
The test are was cleared of hair. A filter paper (2 x 4cm) saturated with vaseline only was applied to the test area and held in contact by an occlusive dressing for 48 hours.

Challenge: Topical Application
Day 20 - treated and control groups
The flanks of treated and control animals were cleared of hair. In treated and control animals a filter paper (2x2cm) saturated with 0.1% test substance in vaseline was applied to the left flank of the animals and a filter paper (2x2cm) with the vaseline only was applied to the right flank. The patches were held in contact by an occlusive dressing for 24 hours.

Clinical signs alongwith mortality and viability were monitored daily, bodyweights were measured at T0 and 24h and skin reactions were measured at 24, 48 and 72 hours after the intradermal injection, at 24, 48 and 72 hours after the topical application and after 48 and 72 hours after challenge.
Challenge controls:
The flanks of treated and control animals were cleared of hair. In treated and control animals a filter paper (2x2cm) saturated with 0.1% test substance in vaseline was applied to the left flank of the animals and a filter paper (2x2cm) with the vaseline only was applied to the right flank. The patches were held in contact by an occlusive dressing for 24 hours.
Positive control substance(s):
yes
Remarks:
Benzocaine (ethyl p-aminobenzoate)

Results and discussion

Positive control results:
Reliability of the experimental technique is checked every 6 months with benzocaine. For intradermal application the 5% concentration in olive oil was used and the challenge exposuer was performed in the same concentration of vaseline. The result of the last experiment (March 2006) produced a positive skin reaction in 4 animals out of a total of 10 animals. The 40% result is satisfactory as the guideline states a minimum number of animals with a positive skin reaction is expected to be 30%.

Any other information on results incl. tables

Bodyweight 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

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The potential of Potassium permanganate to induce delayed contact hypersensitivity (skin sensitisation) was investigated in a Manusson and Kligman maximisation study using 35 albino guinea pigs (12 males and 23 females). The concentrations of the test material used for induction and challenge applications were based on the results of a preliminary study. The experiment consisted of three phases: two induction phases – topical application and the challenge phase. The results of an acceptable positive control study are also reported, confirming the sensitivity of the assay. Induction was performed on 20 test animals using 3 pairs of injections containing 1% test substance in water. Then at day 6, a 48-hour semi-occlusive application of 10% test material in Vaseline was applied. Ten control animals were similarly treated, using vehicle. Two weeks following the final induction application, all test and control animals were challenged using a 24- hour occlusive application of the test material (0.1% test substance in vaseline). Dermal reactions were assessed at 24, 48 and 72 hours following patch removal. No dermal reactions were seen in test or control animals. No evidence of sensitisation was seen under the conditions of this study.
Executive summary:

The potential of Potassium permanganate to induce delayed contact hypersensitivity (skin sensitisation) was investigated in a Manusson and Kligman maximisation study using 35 albino guinea pigs (12 males and 23 females). The concentrations of the test material used for induction and challenge applications were based on the results of a preliminary study. The experiment consisted of three phases: two induction phases – topical application and the challenge phase. The results of an acceptable positive control study are also reported, confirming the sensitivity of the assay. Induction was performed on 20 test animals using 3 pairs of injections containing 1% test substance in water. Then at day 6, a 48-hour semi-occlusive application of 10% test material in Vaseline was applied. Ten control animals were similarly treated, using vehicle. Two weeks following the final induction application, all test and control animals were challenged using a 24- hour occlusive application of the test material (0.1% test substance in vaseline). Dermal reactions were assessed at 24, 48 and 72 hours following patch removal. No dermal reactions were seen in test or control animals. No evidence of sensitisation was seen under the conditions of this study.