Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 September 1991 to 13 January 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study in accordance with internationally recognised guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): ADK STAB PEP-36
- Physical state: White powder
- Purity: >99%
- Lot/batch No.: L-10181
- Expiration date of the lot/batch: 1 August 1992
- Storage condition of test material: At room temperature (approximately 20°C)

In vivo test system

Test animals

Species:
guinea pig
Strain:
Himalayan
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: biological research laboratory
- Age at study initiation: 8 weeks (including acclimatisation)
- Weight at study initiation: pre-test 320 - 344 g, start of acclimatisation control and test group 302 - 371 g
- Housing: individually in Makrolon type-3 cages (size: 27 x 42 x 15 cm) with standard softwood bedding
- Diet: pelleted standard guinea pig breeding/maintenance diet, ad libitum
- Water: community tap water, ad libitum. Once weekly additional supply of ascorbic acid (1g/l) via drinking water
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12

IN-LIFE DATES: From: 7 October 1901 To: 31 October 1991

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: vaselinum album
Concentration / amount:
Pre-test:
1) Intradermal: 5, 3 and 1% in ethanol
2) Epidermal: 25, 15, 10, 5 % in vaselinum
Main test:
Intradermal: 5% in ethanol
Epidermal: 25% in vaselinum
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: vaselinum album
Concentration / amount:
Pre-test:
1) Intradermal: 5, 3 and 1% in ethanol
2) Epidermal: 25, 15, 10, 5 % in vaselinum
Main test:
Intradermal: 5% in ethanol
Epidermal: 25% in vaselinum
No. of animals per dose:
Number of animals in test group: 20
Number of animals in negative control group: 10
Details on study design:
RANGE FINDING TESTS:
To identify irritant concentrations for induction phase of main study and non-irritant concentration by topical route for challenge application.
Intradermal injection - 5, 3 and 1% in ethanol. Dermal reactions assessed after 24 hours. 5% test substance in ethanol was selected for intracutaneous induction in the main study.
Epidermal application - 25, 15, 10 and 5%. Assessed for erythema and oedema 24 hours and 48 hours after exposure. 25% test substance in ethanol was selected for the main study.

MAIN STUDY
A. INDUCTION EXPOSURE
Intradermal
- No. of exposures: 6 exposure sites (3 pairs) per animal
- Exposure period: 7 days

- Test groups:
i)) Freund's complete adjuvant 50:50 with physiological saline.
ii) The test article, diluted to 5% with ethanol
iii) The test article diluted to 5% by emulsion in a 50:50 mixture of Freund's complete adjuvant and physiological saline.

- Control group:
1) Freund's complete adjuvant 50:50 with physiological saline.
2) Ethanol.
3) Freund's complete adjuvant 50:50 with physiological saline.

- Site: scrapular region
- Frequency of applications: 1
- Duration: 7 days
- Concentrations: 5 % test substance in ethanol

Epidermal
On day 7 of the test and approximately 24 hours prior to the epidermal application the scapular area (approximately 6 x 8 cm) was clipped, shaved free of hair and the test areas were pre-treated with 10% Sodium-Lauryl-Sulfate (SLS) in petrolatum oil because no primary irritation concentration could be determined in the corresponding pre-test. The SLS was massaged into the skin with a glass rod without bandaging. This SLS-concentration enhances sensitization by provoking a mild inflammatory reaction.

On day 8 a 2 x 4 cm patch of filter paper was covered with a thin layer of the selected test article concentration (25% in vaselinum album) and placed over the injection sites of the test animals. The patch was covered by aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The dressings were left
in place for approximately 48 hours. The epidermal application procedure described ensured intensive contact of the test article.

The guinea-pigs of the control group were treated as described above with the omission of test article (vaselinum album only).
Reaction sites were assessed for erythema and oedema 24 and 48 hours after removal of the dressing, using the numerical grading system according to Draize.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: two weeks after epidermal exposure
- Exposure period: 24 hours
- Test groups:
i) non-irritant concentration of the test article (25% in vaselinum album
ii) vaselinum album only
- Control group: treated in the same way as the test groups described above.
- Site: 5 x 5 cm area on the left and right flank
- Concentrations: 25%
- Evaluation (hr after challenge): 24 and 48 hours

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
20
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: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
20
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: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
20
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: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
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: negative control. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
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: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
10
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: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
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: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.

Any other information on results incl. tables

Maximum concentration not causing irritating effects in preliminary test: 25 % Signs of irritation during induction: mild erythema and edema
Evidence of sensitisation of each challenge concentration:
A 25 %  0
A 0 %   0

Applicant's summary and conclusion

Interpretation of results:
other: not classified
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
From the results described above no allergenic potency of the test substance was concluded. The results were interpreted according to the rating of Magnusson and Kligman (1969).