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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:
7 May - 1 Jun 1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
adopted 19. Sept. 1984
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Test was done before LLNA as first-choice method for in-vivo testing was set into force.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): T-4113
- Physical state: white powder
- Analytical purity: > 96%
- 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: D. Hall, Newchurch, Staffordshire, England
- Age at study initiation: 6 to 7 weeks of age
- Weight at study initiation: 327 - 389 g
- Housing: in groups of ten in suspended cages with wire mesh floors
- Diet: vitamin C-enriched guinea pig diet F.D.1., ad libitum; hay was given weekly
- Water: ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 30-70
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 7 May To 01 June 1991

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Alembicol D
Concentration / amount:
Induction: intradermal - 2.5% w/w; epicutaneous - 70% w/w
Challenge: epicutaneous - 25 and 50% w/w
Challengeopen allclose all
Route:
epicutaneous, semiocclusive
Vehicle:
other: Alembicol D
Concentration / amount:
Induction: intradermal - 2.5% w/w; epicutaneous - 70% w/w
Challenge: epicutaneous - 25 and 50% w/w
No. of animals per dose:
10 (controls), 20 (in test group)
Details on study design:
RANGE FINDING TESTS: The intradermal and topical irritancy of a range of dilutions of T-4113 was investigated to identify where possible (a) irritant test concentrations suitable for induction phase of the main study and (b) non-irritant concentrations by the topical route of administration for the challenge phase. Concentration ranges tested in two animals each were: intradermal - 0.1, 0.25, 0.5, 1.0, 2.5, 5.0, 7.5, 10.0% (w/w); topical - 10, 30, 50, 70% (w/w)

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single intradermal injection and 48 hours topical application
- Test groups:
Intradermal - 3 pairs of injections: (1) a 1:1 mixture (v/v) Freund's Complete adjuvant (FCA)/water; (2) test substance in Alembicol D; (3) test substance in a 1:1 mixture of Freund's complete adjuvant and Alembicol D
Epicutaneous - test substance in Alembicol D
- Control group:
Intradermal - 3 pairs of injections: (1) a 1:1 mixture (v/v) FCA/water; (2) Alembicol D; (3) 1:1 mixture Freund's Complete adjuvant and Alembicol D
Epicutaneous - Alembicol D
- Site: scapular region, intradermal and epicutaneous
- Frequency of applications: every 7 days
- Duration: Days 0-9
- Concentrations: intradermal 2.5% w/w, epicutaneous 70% w/w

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 21
- Exposure period: 24 hours
- Test groups: test substance at 25 and 50% w/w in Alembicol D
- Control group: test substance at 25 and 50% w/w in Alembicol D
- Site: anterior and posterior site on the left flank
- Concentrations: 25 and 50% w/w
- Evaluation (hr after challenge): 24, 48 and 72 hours

OTHER:
Justification for choice of vehicle: Alembicol D# is a product of coconut oil. By fractionating the fatty acids of coconut oil and re-esterifying with glycerine, a Medium Chain Triglyceride (MCT) Oil is produced. Alembicol D is such an oil. It is advantageous in this type of test in that it is stable, has a low viscosity and surface tension and is easily absorbed into the skin. These properties make it ideal as a vehicle for intradermal injections and topical applications.
Positive control substance(s):
yes
Remarks:
Formalin (checked periodically)

Results and discussion

Positive control results:
Formalin is used as positive control and checked regularly to test the stability of the assay; concentrations used: intradermal - 0.05 -0.1%; epicutaneous - 5 -10%; challenge - 1 -5%; usually 50 - 100% of the tested animals showed sensitizing reactions in all studies conducted

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
intradermal induction: 2.5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: intradermal induction: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge..
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
intradermal induction: 2.5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: intradermal induction: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge..
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
intradermal induction: 2.5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving Alembicol D. Slight erythema following topical application. No dermal reactions at challenge.
Remarks on result:
other: see Remark
Remarks:
Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: intradermal induction: 2.5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving Alembicol D. Slight erythema following topical application. No dermal reactions at challenge..
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
intradermal induction: 2.5%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: intradermal induction: 2.5%. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge..
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
intradermal induction: 2.5%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge.
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: intradermal induction: 2.5%. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge..
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
intradermal induction: 2.5%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge.
Remarks on result:
other: see Remark
Remarks:
Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: intradermal induction: 2.5%. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: Skin reactions at induction: Necrosis at intradermal application sites receiving FCA and slight irritation at sites receiving 2.5% test substance in Alembicol D. Slight erythema following topical application. No dermal reactions at challenge..

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
DSD: not classified
CLP: not classified