Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well conducted GLP study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report Date:
2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Test article name : Emulsamine AT-O
Chemical name : alkylamidoamine
Batch number: 6514
Purity: > 95%
Origin: Feuchy plant
Expiration date of the lot/batch: july 2003
Physical state: dark brown liquid
Storage condition of test material: at room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, l'Arbresle, France
- Age at study initiation: 1-2 months old
- Weight at study initiation: 389+/-23g for the males and 365+/-22g for the females
- Housing: individual polycarbonate cage with stainless steel lid (48cm*27cm*20cm)
- Diet : free access to 106 pelleted diet (UAR, Villemoisson, Epinay sur Orge, France)
- Water : drinking water filtered by a FG Millipore membrane (0.22 micron), ad libitum
- Acclimation period: at least 5 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-2
- Humidity (%): 30 to 70
- Air changes (per hr): approximately 12 cycles/hour of filtered, non recycled air
- Photoperiod (hrs dark / hrs light): 12h/12h

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: intradermal injections : corn oil; topical applications: a 80/20 (w/w) mixture ethanol and purified water(induction phase); acetone (challenge application).
Concentration / amount:
intradermale route: 0.1% (w/w) in corn oil; cutaneous route: 10% (w/w) in ethanol/water (induction phase); 1% (w/w) (challenge application) in acetone
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: intradermal injections : corn oil; topical applications: a 80/20 (w/w) mixture ethanol and purified water(induction phase); acetone (challenge application).
Concentration / amount:
intradermale route: 0.1% (w/w) in corn oil; cutaneous route: 10% (w/w) in ethanol/water (induction phase); 1% (w/w) (challenge application) in acetone
No. of animals per dose:
control group : 10 animals (5 males and 5 females)
treated group : 20 animals ( 10 males and 10 females)
Details on study design:
RANGE FINDING TESTS:
4 males and 4 females

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 30
- Exposure period: -
- Test groups: test substance in corn oil
- Control group: corn oil only
- Site: Right and left flank
- Duration: 0-8 day

B. CHALLENGE EXPOSURE
- No. of exposures: 30
- Day of challenge: 22
- Exposure period: 24h
- Test groups: test substance
- Control group: test substance
- Site: right flank for the test substance and left flank for the vehicle
- Concentrations: 1% (w/w) in acetone
- Evaluation (hr after challenge): 24, 48

Scale :
No visible change : 0
Discrete or patchy erythema : 1
Moderate and confluent erythema : 2
Intense erythema : 3
Challenge controls:
5 males and 5 females
50% Freund’s complete adjuvant in 0.9% NaCl; vehicle (only); vehicle at 50% in the mixture Freund’s complete adjuvant / 0.9% NaCl (50/50)
Positive control substance(s):
yes
Remarks:
mercaptobenzothiazole

Results and discussion

Positive control results:
Sensitization response in 100% animals to mercaptobenzothiazole

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no clinical reactions
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no clinical reactions.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
1%
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
discrete erythema (grade 1)
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1%. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: discrete erythema (grade 1).
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
Dryness of the skin
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: Dryness of the skin.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
1%
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
Dryness of the skin
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1%. No with. + reactions: 3.0. Total no. in groups: 20.0. Clinical observations: Dryness of the skin.

Any other information on results incl. tables

In the treated group, a discrete erythema (grade 1) was noted in 1/20 animals at the 24-hour reading. No erythema was recorded at the 48-hour reading.

Dryness of the skin was observed in 3/20 animals of the treated group and in 1/10 animal of the control group at the 48-hour reading.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Under our experimental conditions and according to the maximization method of Magnusson and Kilgman, the test substance does not induce delayed contact hypersensitivity in guinea pigs.
Executive summary:

The potential of the test substance to induce delayed contact hypersensitivity was evaluated in guinea pigs according to the maximization method of Magnusson and Kligman and to OECD (No. 406, 17th July 1992) and EC (96/54/EEC, B.6, 30 July 1996) guidelines.

The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

Thirty guinea pigs were allocated to two groups: a control group of five males and fives females and a treated group of ten males and ten females.

On day 1, three pairs of interdermal injections were performed in the interscapular region of all animals:

-         Freund’s complete adjuvant (FCA) diluted in 50% (v/v) with 0.9% NaCl (both groups)

-         Test substance at the concentration of 0.1% in corn oil (treated group) or vehicule alone (control group)

-         Test substance at the concentration of 0.1% in a mixture FCA / 0.9% NaCl  (50/50, v/v) (treated group) or vehicle at the concentration of 50% (v/v) in a mixture FCA / 0.9% NaCl (50/50, v/v) (control group).

On day 8, the animals of the treated group received a topical application of the test substance at the concentration of 10% (w/w) in ethanol water (80/20) to the same test site, which was then covered by an occlusive dressing for 48 hours. The animals of the control group received an application of vehicle under the same experimental conditions.

On day 22, all animals of both groups were challenged by a cutaneous application of the test substance at the concentration of 1% (w/w) in acetone to the right flank. The test substance was maintained under an occlusive dressing for 24 hours; The vehicle was applied to the left flank under the same experimental conditions.

Skin reactions were evaluated approximately 24 and 48 hours after removal of the dressing.

At the end of the study, animals were killed without examination of internal organs.

No skin samples were taken from the challenge application sites.

No clinical signs an no deaths were noted during the study.

After the challenge application, no cutaneous reactions were observed in the animals of the control group.

In the treated group, a discrete erythema was noted at the 24-hour reading in 1/20 animals. No erythema was recorded at the 48-hour reading.

Dryness of the skin was observed in 3/20 animals of the treated group and in 1/10 animal of the control group at the 48-hour reading.

Under the experimental conditions and according to the maximization method of Magnusson and Kilgman, the test substance does not induce delayed contact hypersensitivity in guinea pigs.

According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations), the test substance should not be considered as a skin sensitizer.