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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:
04 April, 1995 - 05 May, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
(1992)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
(1992)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: "Allergic Contact Dermatitis in the Guinea-Pig: Identification of Contact Allergens" Magnusson B. Kligman A.M., 1970 published by C.C. Thomas, Springfield, Illinois, USA.
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Adequate data from a reliable GPMT study were already available.

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
422-150-1
EC Name:
-
Molecular formula:
Hill formula: Mg4.3 Al2(OH)12.6 (CO3)0-0.75 (ClO4)0.5-2.0 . (0-5)H2O CAS formula: Mg4.3 Al2(OH)12.6 (CO3)0-0.75 (ClO4)0.5-2.0 . (0-5)H2O
IUPAC Name:
Aluminium-magnesium-carbonate-hydroxide-perchlorate-hydrate
Test material form:
solid: particulate/powder
Details on test material:
- Appearance: white powder
- Storage condition of test material: at room temperature in the dark

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany
- Age at study initiation: approx. 6-7 weeks
- Weight at study initiation: 446-556 g
- Housing: Group housing of 2 animals, except for 1 control animal, per labelled metal cage with wire-mesh floors.
- Diet: Free access to standard guinea pig diet, including ascorbic acid (1600 mg/kg) (LC 23-B, pellet diameter 4mm, Hope Farms, Woerden, The Netherlands)
- Water: Free access to tap water, diluted with decalcified water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): approx. 15
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: distilled water
Concentration / amount:
- 25% for the intradermal induction
- 50% for the epidermal induction
- 0, 10, 25 and 50% for the challenge
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: distilled water
Concentration / amount:
- 25% for the intradermal induction
- 50% for the epidermal induction
- 0, 10, 25 and 50% for the challenge
No. of animals per dose:
Number of animals in test group: 10
Number of animals in negative control group: 5
Details on study design:
RANGE FINDING TESTS (4 animals, age: approx. 6-7 weeks)
For the intradermal induction exposure (2 animals):
- Concentration: 10 and 25%; and 2 and 5%
- Amounts: 0.1 mL. Each of the 2 animals received 2 different concentrations in the right and left clipped shoulder regions.
- Scoring: the skin reactions were scored 24 and 48 hours after application

For the epicutaneous induction, using Scotchpak-non-woven patch (the same animals as above):
- Concentration: 50 and 25%
- Amount: 0.5 mL. Each of the 2 animals received a differerent concentration to the shaved flank of the animal.
- Exposure period: 24 hours (occlusive)
- Scoring: the skin reactions were scored 24 and 48 hours after patch removal

For the epicutaneous induction, using square chambers (2 animals):
- Concentration: 50, 25, 10 and 5%
- Amount: 0.05 mL were applied occlusively on a shaved flank of each of the 2 animals.
- Exposure period: 24 hours (occlusive)
- Scoring: the skin reactions were scored 24 and 48 hours after patch removal

MAIN STUDY
Test animals: 10
Control animals: 5

A. INDUCTION EXPOSURE
- No. of exposures: 2
1) Intradermal injections on day 1:
- Concentration: 25%
- Site: clipped scapular region. One of each pair was on each side of the midline and from cranial A) to caudal C):
Three pairs of intradermal injections:
1) 0.1 mL: FCA (50% in water)
2) 0.1 mL: test substance at a 25% concentration in distilled water (control animals: 0.1 mL distilled water)
3) 0.1 mL: 50% test substance in distilled water + 50% FCA (undiluted) (control animals: 50% with distilled water + 50% FCA (undiluted))
-Readings: on day 3 (48 hrs after the injections). Irritating results were observed in the test groups, in each of the 3 sites.
- Use of SDS (all animals): on day 8, 24 hrs before the topical induction application, the scapular area was clipped and subsequently rubbed with 10% SDS in vaseline using a spatula.

2)Topical application on day 9:
- Concentration: 50%
- Amount: 0.5 mL (control animals: 0.5 mL of distilled water), on the SDS treated area
- Exposure period: 48 hours (occlusive)
- Readings: scores were rated directly after patch removal (skin cleaned of residual test substance)

B. CHALLENGE EXPOSURE (all animals)
- Concentration: 0, 10, 25 and 50%
- Day of challenge: day 22 (13 days after the start of the 2nd induction)
- Exposure period: 24 hours (occlusive)
- Site: flank
- Amount: 0.5 mL
- Readings: scores were rated 24 and 48 hours after patch removal
Challenge controls:
Not applicable.

Positive control substance(s):
yes
Remarks:
(the results of the latest reliability check, performed in December 1994 with Alpha-hexylcinnamicaldehyde, tech. 85%, are reported)

Results and discussion

Positive control results:
The latest reliability check (performed less than 6 months ago) shows a sensitisation rate of 90%. See the attached background material.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50 %
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 2.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25 %
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 %. No with. + reactions: 2.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10 %. No with. + reactions: 2.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
10%, 25% and 50%
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
10%, 25% and 50%
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation

Any other information on results incl. tables

- In the pre-liminary test, discrete or patchy erythema observed at intradermal induction with 25% test substance in the animal. Therefore 25% for the intradermal induction was used.

- Maximum concentration not causing irritating effects in the preliminary epidermal test was 50 %.

Main test:

Signs of irritation after induction:
After intradermal injection (sites without FCA): On day 3 moderate/severe erythema with or without necrosis was observed in all animals (experimental). On day 11, 9/10
animals showed eschar formation and 10/10 animals showed moderate/severe erythema. Control animals showed no reactions.

Signs after epidermal application: 9/10 experimental animals showed very slight to severe erythema. 2/10 animals also showed eschar/crust formation.
1/5 control animals showed very slight erythema.

Challenge:

No skin reactions were evident after the challenge exposure.

Two animals showed discrete or patchy erythema in repsonse to the 50, 25 and 10% test substance concentrations 48 hours after patch removal. One other animal showed scaliness is response to the 50% concentration.

Two of the ten experimental animals were considered indicative of sensitisation.

One control animal died by accident on day 22. Pink discolouration of the lungs was found at post mortem macroscopic examination of this animal.

No further mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study during the study period.

The average body weight of the experimental animals and the control animals was within an acceptable range at the beginning and the end of the study.

Applicant's summary and conclusion

Interpretation of results:
other: not sensitising
Remarks:
According to Regulation (EC) No. 1272/2008 and its amendments.
Conclusions:
In an adjuvant type guinea pig test method for skin sensitisation according to OECD 406 guideline and GLP principles, two of the ten experimental animals showed positive reactions during challenge with Alcamizer 5.

Executive summary:

Alcamizer 5 was tested in an adjuvant type guinea pig test method for skin sensitisation according to OECD 406 guideline and GLP principles.

After intradermal injection (sites without FCA) on day 3, moderate/severe erythema with or without necrosis was observed in all animals (experimental). On day 11, nine of the ten animals showed eschar formation and  all ten animals showed moderate/severe erythema. Control animals showed no reactions. After epidermal application, nine of the ten experimental animals showed very slight to severe erythema. Two of the ten animals also showed eschar/crust formation. One of the five control animals showed very slight erythema.

One control animal died by accident on day 22. Pink discolouration of the lungs was found at post mortem macroscopic examination of this animal. No further mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study during the study period.

After the challenge, no skin reactions were evident after the challenge exposure.

Two animals showed discrete or patchy erythema in repsonse to the 50, 25 and 10% test substance concentrations 48 hours after patch removal. One other animal showed scaliness is response to the 50% concentration. These results lead to a sensitisation rate of 20%.

The latest reliability check (performed less than 6 months ago) shows a sensitisation rate of 90%.

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