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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:
1986-01-03 to 1986-02-09
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Guideline conform study, conducted under GLP principles, with minor deficiencies when compared to contemporary standards: the purity and the stability of the test material were not stated in the study report. The guinea pig strain used in these laboratories is reported to have been shown to produce a satisfactoy sensitisation response using positive sensitisers. However, it was not stated in the study report which positive sensitisers were tested or when the last test was conducted. According to the guideline, the test have be conducted at least every six months.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986
Report Date:
1986

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1981-05-12
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The conduct of this study was prior to the establishment of the LLNA method (OECD 429).

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Rezal 36 GP powder
- Label: Ref.QC 1433/569 REZAL 36 GP Aluminium Zirconium Tetra Chlorhydrex Gly powder
- Physical state: white powder
- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: approximately seven to eleven weeks old
- Weight at study initiation: 365 - 410 g
- Housing: the animals were housed in groups of up to four in solid floor polypropylene cages furnished with softwood shavings.
- Diet (ad libitum): Guinea Pig FD1 Diet, Special Diet Services Limited, Witham, Essex, U.K.
- Water (ad libitum): mains tap water
- Acclimation period: minimum of five days

ENVIRONMENTAL CONDITIONS
- Temperature: 17 - 23°C
- Relative humidity: 45 - 65%
- Air exchanges: approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Intradermal induction: 0.5% (w/v) in distilled water
Topical induction: 50% (w/w) in distilled water
Topical challenge: 50% (w/w) in distilled water
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Intradermal induction: 0.5% (w/v) in distilled water
Topical induction: 50% (w/w) in distilled water
Topical challenge: 50% (w/w) in distilled water
No. of animals per dose:
Sighting study: 8 animals (intradermal 4 animals; topical 4 animals)
Main study:
test group: 20 animals
vehicle control group: 10 animals
Details on study design:
RANGE FINDING TESTS:
The dose levels for each of the three stages of the main study were determined by a 'sighting study' in which two groups of four guinea pigs were used. The procedure was as follows:
i) Intradermal injection (induction): dilutions of the test material in distilled water (0.5, 0.1, 1 and 5% concentrations) were tested to determine the highest level that could be well tolerated both locally and systemically.
ii) Topical application (induction): dilutions of the test material in distilled water (25% and 50% concentrations) were tested to determine the highest level which did not produce excessive inflammation in animals injected with Freund's Complete Adjuvant (Difco Laboratories, Michigan, U.S.A.) at least seven days previously.
iii) Topical application (challenge): dilutions of the test material in distilled water (25% and 50% concentrations) were tested to determine the highest level which did not produce inflammation or irritation in animals injected with Freund's Complete Adjuvant at least seven days previously.

Results:
- Intradermal sighting test: the concentration of the test material to be used for the intradermal induction stage of the main study was selected to be 0.5% (w/v) in distilled water.
- Topical sighting study: the concentration of the test material to be used for the main study was selected as follows:
topical induction = 50% (w/w) in distilled water
topical challenge = 50% (w/w) in distilled water


MAIN STUDY
A. INDUCTION EXPOSURE
1) Test animals:
The hair was removed from an area approximately 40 mm X 60 mm on the shoulder region of each animal with veterinary clippers and a row of three injections (0.1 mL each) was made on each side of the midline. The injections were:
i) Freund's Complete adjuvant plus distilled water in the ratio 1:1.
ii) a 0.5% (w/v) dilution of test material in distilled water.
iii) a 0.5% (w/v) dilution of test material in a 1:1 preparation of Freund's Complete Adjuvant plus distilled water

One week later, the shoulder region was clipped again and treated with a topical application of the test material formulation (50% w/w in distilled water). the test material formulation (0.2 - 0.3 mL) was applied on filter paper (WHATMAN No. 4: approximate size 40mm X 20 mm) which was held in place with two strips of waterproof adhesive strapping (SLEEK) in the form of a cross and covered with an overlapping length of aluminium foil. The patch and foil were further secured by a strip of elastic adhesive bandage (ELASTOPLAST: approximate size 250 mm X 35 mm) wound in a double layer around the torso of each animal. This occlusive dressing was kept in place for 48 hours.

2) Control animals:
Intradermal injections were administered using an identical procedure to that used for the test animals, except that the injections were:
i) Freund's Complete Adjuvant plus distilled water in the ratio 1:1.
ii) Distilled water
iii) Freund's Complete Adjuvant plus distilled water in the ratio 1:1.
The topical applications followed the same procedure as for the test animals except that the vehicle alone was applied to the filter paper.

B. CHALLENGE EXPOSURE
Two weeks after the topical inductions, an area, approximately 50 - 70 mm X 50 mm on both flanks of each animal, was clipped free of hair with veterinary clippers.
A quantity of 0.1 - 0.2 mL of the test material formulation (50% w/w in distilled water) was applied to the shorn right flank of each animal on a 20 mm X 20 mm square of filter paper (WHATMAN No.4) which was held in place by two strips of waterproof adhesive strapping (SLEEK) in the form of a cross. The vehicle alone was similarly applied to the left shorn flank. The patches were occluded with an overlapping length of aluminium foil and secured by a strip of elastic adhesive bandage (ELASTOPLAST: approximate size 250 mm X 75 mm) wound in a double layer around the torso of each animal.

After 24 hours, the dressing was carefully cut using blunt-tipped scissors, removed and discarded. the position of the sites were identified by using a black indelible pen. After a further 24 and 48 hours, any erythematous reactions were quantified using a slightly modified Magnusson and Kligman scale.

OBSERVATIONS:
The bodyweight of each animal was recorded at the start and end of the study.
Challenge controls:
Vehicle control group: distilled water
10 females were used in the vehicle control group.
Positive control substance(s):
yes
Remarks:
The strain used in these laboratories has been shown to produce a satisfactoy sensitisation response using positive sensitisers (NOTE: it was not stated in the study report which positive sensitisers were tested or when the last test was conducted).

Study design: in vivo (LLNA)

Statistics:
no data

Results and discussion

Positive control results:
The strain used in these laboratories has been shown to produce a satisfactory sensitisation response using positive sensitisers.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50% (w/w) of the test material in distilled water
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No adverse skin reactions were observed at the test material site.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50% (w/w) of the test material in distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No adverse skin reactions were observed at the test material site..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50% (w/w) of the test material in distilled water
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No adverse skin reactions were observed at the test material site.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50% (w/w) of the test material in distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No adverse skin reactions were observed at the test material site..
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
distilled water
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No adverse skin reactions were observed at the vehicle control site.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No adverse skin reactions were observed at the vehicle control site..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
distilled water
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No adverse skin reactions were observed at the vehicle control site.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No adverse skin reactions were observed at the vehicle control site..
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50% (w/w) of the test material in distilled water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No adverse skin reactions were observed at the test material site.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50% (w/w) of the test material in distilled water. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No adverse skin reactions were observed at the test material site..
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50% (w/w) of the test material in distilled water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No adverse skin reactions were observed at the test material site.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% (w/w) of the test material in distilled water. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No adverse skin reactions were observed at the test material site..
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
distilled water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No adverse skin reactions were observed at the vehicle control site.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No adverse skin reactions were observed at the vehicle control site..
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
distilled water
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No adverse skin reactions were observed at the vehicle control site.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No adverse skin reactions were observed at the vehicle control site..

Any other information on results incl. tables

OBSERVATIONS:

- Bodyweights: bodyweight gains of guinea pigs in the test group between day 0 and 24 were comparable to those in the control group over the same period.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material, Rezal 36 GP powder, was found to be a non-sensitiser to guinea pig skin.
According to the EC-Commission directive 67/548/EEC and its subsequent amendments, the test substance is not a skin sensitizer.
According to the EC-Regulation 1272/2008 and its subsequent amendments, the test item is not a skin sensitizer.