Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 10 December 1991 and 4 January 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
EEC Directive 84/449/EEC, Annex V, Method B6
Deviations:
yes
Remarks:
0.2 mL of sodium lauryl sulphate instead of 0.5 mL recommended in the guideline
Principles of method if other than guideline:
not applicable
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): P-#620

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: 302 to 367 g
- Housing: in groups of 10 in suspended metal cages with wire mesh floors
- Diet (e.g. ad libitum): vitamin C-enriched guinea-pig diet F.D.1 ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

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

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Alembicol D
Concentration / amount:
Concentration of test material and vehicle used at induction:
- Intradermal injection: 20% in Alembicol D or in 50:50 mixture FCA:Alembicol D.
-Topical application: As supplied

Concentration of test material and vehicle used for each challenge:
- Topical application: As supplied
- Topical application: 50% v/v in Alembicol D
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Alembicol D
Concentration / amount:
Concentration of test material and vehicle used at induction:
- Intradermal injection: 20% in Alembicol D or in 50:50 mixture FCA:Alembicol D.
-Topical application: As supplied

Concentration of test material and vehicle used for each challenge:
- Topical application: As supplied
- Topical application: 50% v/v in Alembicol D
No. of animals per dose:
20 test animals, 10 control animals
Details on study design:
RANGE FINDING TESTS:
- Intradermal injections: 1, 2.5, 5, 7.5, 10, 20, 30, and 40% in Alembicol D. using 4 animals
- Topical application: 50, 60, 75 and 100% in Alembicol D. using 4 animals

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2, intradermal injections and topical application
- Exposure period: 48h for topical application
- Test groups: 3 pairs of intradermal injection (0.1 mL) on Day 0 after shaving. As the substance is not a skin irritant, 6 days after the injections, after close-clipping, the site was pre-treated with 0.2 mL of 10% w/w sodium lauryl sulphate in petrolatum. 24 hours later, 48 hours occlusive application on Day 7 after shaving (Whatman No. 3 paper covered by a length of impermeable plastic adhesive tape and firmly secured by elastic adhesive bandage wound around the torso of the animal and fiwed "Sleek" impervious plastic adhesive tape).
- Control group: similarly treated with the exception that the test substance was omitted
- Site: 2 x 4 cm area of dorsal skin on the scapular region
- Concentrations: 20 % w/v in Alembicol D for intradermal exposure and as supplied for topical exposure

B. CHALLENGE EXPOSURE
- No. of exposures: 1 topical exposure
- Day(s) of challenge: 2 weeks after the topical induction application
- Exposure period: 24 hours
- Test groups: Hair was removed by clipping and then shaving. A 20 x 20 mm patch of Whatman No. 3 paper was used for each site. The patches were sealed to the flank for 24 hours under strips "Blenderm" covered by "Elastoplast" wound around the trunk and secured with "Sleek".
- Control group: similarly treated with the exception that the test substance was omitted
- Site: area of the left flank
- Concentrations: 100 % (anterior site of the flank) and 50 % (posterior site of the flank) in Alembicol D.
- Evaluation (hr after challenge): 24, 48 and 72 h after removal of the patches
Challenge controls:
The control test animals were challenged topically two weeks after the topical induction application.
Positive control substance(s):
no
Remarks:
the sensitivity of the guinea-pig is checked periodically with formalin

Results and discussion

Positive control results:
The sensitivity of the guinea-pig is checked periodically with formalin

In vivo (non-LLNA)

Resultsopen allclose all
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:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50 %
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: 50 %. 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:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50 %
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: 50 %. No with. + reactions: 0.0. Total no. in groups: 20.0.
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:
1st reading
Hours after challenge:
24
Group:
negative control
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: negative control. Dose level: 50 %. 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.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50 %
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: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0.

Any other information on results incl. tables

Signs of irritation during induction:
Intradermal injection: Necrosis was recorded at sites receiving Freund's complete Adjuvant in test and control animals.
Irritation was seen in test animals at sites receiving the substance, 20% v/v in Alembicol D.
Topical application: Very slight to moderate erythema in test animals with the substance as supplied.

Evidence of sensitisation of each challenge concentration: None.

Other observations:

No signs of ill health or toxicity were recorded in any animals during the observation period. Bodyweight gain appears normal in all animals.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Under the test conditions, P-#620 is not classified according to the Annex VI of the Regulation (EC) No 1272/2008 (CLP) and the Annex VI of the Directive 67/548/EC.
Executive summary:

In a dermal sensitization study performed according to the EU test method B.6 and in compliance with GLP, P-#620 (89.01% a.i.) was tested in female Dunkin/Hartley guinea-pigs using the Guinea-Pig Maximisation Test method (20 treated animals + 10 controls).

 

The preliminary study determined the lowest irritant test substance concentration used at induction phases and the highest non-irritant test substance concentration used at challenge.

P-#620 diluted in Alembicol D at 20% (v/v) was administered by intradermal injection for the induction. As the substance is not a skin irritant, 6 days after the injections the site was pre-treated with 0.2 mL of 10% w/w sodium lauryl sulphate in petrolatum. Then topical induction was performed with P-#620 as supplied on day 7.

For the challenge, P-#620 was tested at 100% and 50% (v/v in Alembicol D.).

 

The sensitivity of the guinea-pig is checked periodically at HRC with formalin, a known sensitizer.

 

No signs of ill health or toxicity were recorded in any animals during the observation period.

 

P-#620 did not produce evidence of skin sensitization (delayed contact hypersensitivity).

 

Under the test conditions, P-#620 is not classified according to the annex VI of the Regulation EC No. 1272/2008 (CLP) and of the Directive 67/548/EEC.

This study is considered as acceptable and satisfies the requirement for sensitisation endpoint.