Registration Dossier

Diss Factsheets

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 10-03-1990 to 11-23-1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study - no positive control in the study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
Reliability scoring based on 1981 guideline for test n°406
Deviations:
yes
Remarks:
: no data reported on the positive control in the report
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Reliable results from a guinea pig maximisation test is already available.

Test material

Constituent 1
Reference substance name:
Armeen M2HT
IUPAC Name:
Armeen M2HT
Constituent 2
Reference substance name:
Amines, bis(hydrogenated tallow alkyl)methyl
EC Number:
262-991-8
EC Name:
Amines, bis(hydrogenated tallow alkyl)methyl
Cas Number:
61788-63-4
IUPAC Name:
61788-63-4
Details on test material:
- Name of test material (as cited in study report): Armeen M2HT
- CAS 61788-63-4 described as ditallowmethylamine,hydrogenated
- Physical state: cream coloured waxy solid
- Analytical Purity: 100.2%
- Purity test date: 19 july 1990
- Batch No.: AT08/C-29.06.90
- Storage condition of test material: at room temperature, protected from light

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Olac Limited, Bicester, Oxfordshire, England
- Age before acclimatation period: 4-6 weeks old
- Weight at study initiation: 330-451 g
- Housing: in stainless steel cages with grid floors, 5 animals of the same sexe in each cage
- Diet : ad libitum (samples of diet were analysed to detect contamination by PCB and chlorinated hydrocarbons)
- Water: ad libitum
- Acclimation period: yes but no more precision

ENVIRONMENTAL CONDITIONS
- Temperature : 18-20 °C
- Humidity: below 40 %
- Air changes : 15 per hour
- Photoperiod (hrs dark / hrs light):12/12 (no source of natural light)

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
paraffin oil
Concentration / amount:
- Induction exposure:
intrademal: 10% w/v Test Substance (TS) in vehicle
occlusive epicutaneous: 10% w/v
- Challenge exposure:
occlusive epicutaneous: 3 % w/v and 0.5 % w/v
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
- Induction exposure:
intrademal: 10% w/v Test Substance (TS) in vehicle
occlusive epicutaneous: 10% w/v
- Challenge exposure:
occlusive epicutaneous: 3 % w/v and 0.5 % w/v
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: conducted to define the concentrations to be tested in the main study.

Intradermal exposure:
24 hours before treatment: dorsal region clipped.
Intradermal administration of 0.1 mL of the test substance (TS) at increasing concentrations.
Evaluation of the potential cutaneous reactions 24 and 48 hours and approximately 168 hours after injection.

Epicutaneous exposure:
24 hours before treatment: dorsal region clipped.
0.4 mL of each concentration applied to a gauze patch of approximately 4 cm2 for 48 hours by occlusive dressing (4 concentrations per animal).
Evaluation of the potential cutaneous reactions 24 and 48 hours after patch removal.


MAIN STUDY

A. INDUCTION EXPOSURE
- No. of exposures: 1 intradermal, 1 epicutaneous
- Exposure period: epicutaneous: 48 hours

-> TEST GROUPS:
Intradermal exposure
Three injections of 0.1 mL were injected into each side of the animal as follows:
. Freund's complete adjuvant (FCA) diluted to 50% with distilled water
. TS at 10% w/v in vehicle
. TS at 5% w/v in a 50/50 (v/v) mixture of FCA and distilled water

Epicutaneous exposure
Application of 0.6 mL of TSat 10% w/v in paraffin oil applied to the scapular region and held in place for 48 hours using an occlusive dressing.

-> CONTROL GROUP:
Intradermal exposure
Three injections of 0.1 mL were injected into each side of the animal as follows:
. Freund's complete adjuvant (FCA) diluted to 50% with distilled water
. vehicle
. A mixture 50/50 (v/v) FCA diluted to 50% with distilled water, and vehicle

Epicutaneous exposure
Application of 0.6 mL of vehicle applied to the scapular region and held in place for 48 hours using an ocular dressing.

- Site:
Intradermal exposure
6 injections in the clipped area (4 x 2 cm) in the scapular region

Epicutaneous exposure
4 x 2 cmarea over the scapulae

- Frequency of applications:
One intradermal injection and one epicutaneous application 8 days after on the same site.

- Duration:
Epicutaneous exposure: 48 hours


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
- Exposure period: 24 hours

-> TEST GROUPS:
0.03 mL of the TS at 3% w/v in vehicule to one site and 0.03 mL of the TS at 0.5% w/v in vehicule to a second site on the posterior right flank (occlusiv e epicutaneous application)
0.03 mL of the vehicle on the posterior left flank (occlusive epicutaneous application)

-> CONTROL GROUPS:
Same treatment as test group
- Site: posterior right and left flanks
- Evaluation (hr after challenge): 24 and 48 hours after removal of the dressing

Challenge controls:
Vehicle was applied at the challenge application on the posterior left flank of both test and control groups of animals.
Positive control substance(s):
not specified

Results and discussion

Positive control results:
no data.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
challenged with Armeen M2HT 3%
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: challenged with Armeen M2HT 3%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Challenged with Armeen M2HT 3%
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: Challenged with Armeen M2HT 3%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
Induced with Armeen M2HT and challenged with Armeen M2HT 3%
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
slight erythema
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Induced with Armeen M2HT and challenged with Armeen M2HT 3%. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: slight erythema.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
Induced with Armeen M2HT and challenged with Armeen M2HT 3%
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
slight erythema
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Induced with Armeen M2HT and challenged with Armeen M2HT 3%. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: slight erythema.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
challenged with Armeen M2HT 0.5 %
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: challenged with Armeen M2HT 0.5 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
challenged with Armeen M2HT 0.5 %
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: challenged with Armeen M2HT 0.5 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
Induced with Armeen M2HT and challenged with Armeen M2HT 0.5 %
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
slight erythema
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Induced with Armeen M2HT and challenged with Armeen M2HT 0.5 %. No with. + reactions: 3.0. Total no. in groups: 20.0. Clinical observations: slight erythema.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
Induced with Armeen M2HT and challenged with Armeen M2HT 0.5 %
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
slight erythema
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Induced with Armeen M2HT and challenged with Armeen M2HT 0.5 %. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: slight erythema.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
challenged with paraffin oil
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: challenged with paraffin oil. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
challenged with paraffin oil
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: challenged with paraffin oil. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
induced with Armeen M2HT and challenged with paraffin oil
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: induced with Armeen M2HT and challenged with paraffin oil. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
Induced with Armeen M2HT and challenged with paraffin oil
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: Induced with Armeen M2HT and challenged with paraffin oil. No with. + reactions: 0.0. Total no. in groups: 20.0.

Any other information on results incl. tables

CHALLENGE PHASE: individual dermal responses after removal of occlusive dressings

Group

Sexe

Animal

Topical application of 0.03 ml 3 %w/v Armeen in paraffin oil

Topical application of 0.03 ml 0.5 %w/v Armeen in paraffin oil

Topical application of 0.03 ml paraffin oil

24 hours after removal of dressing

48 hours after removal of dressing

24 hours after removal of dressing

48 hours after removal of dressing

24 hours after removal of dressing

48 hours after removal of dressing

Control

M

601

0

0

0

0

0

0

602

0

0

+-

+-

0

0

603

0

0

0

0

0

0

604

0

0

0

0

+-

+-

605

0

0

0

0

0

0

Control

F

606

0

0

0

0

0

0

607

0

0

0

0

0

0

608

0

0

0

0

+-

+-

609

0

0

0

0

0

0

610

0

0

0

0

0

0

Treated

M

611

+-

0

0

0

0

0

612

+-

0

0

0

0

0

613

0

0

0

+-

0

0

614

+-

+-

+-

+-

+-

+-

615

+-

+-

+-

0

+-

0

616

0

0

0

+-

0

0

617

+-

0

0

0

+-

0

618

0

0

0

0

+-

+-

619

+-

0

+-

0

0

0

620

0

+-

0

+-

+-

0

Treated

F

621

0

0

0

0

0

0

622

0

0

0

0

0

0

623

0

0

0

0

+-

+-

624

+-

0

0

0

0

0

625

+-

1

0

1

+-

+-

626

+-

+-

+-

1

0

0

627

+-

0

+-

0

+-

+-

628

1

0

0

+-

0

0

629

+-

0

0

0

+-

0

630

0

0

0

1

0

0

0: No response

+-: Barely perceptible erythema

1: Slight erythema

2: Moderate erythema

3: Severe erythema

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
In the treated group, following cutaneous reactions were noted:
-Challenge application of 3 % w/v Armeen M2HT in paraffin oil caused slight erythema in 1 out of 20 animals in the treated group after the 24 and the 48-hour readings and no cutaneous reactions in the control group.
-Challenge application of 0.5 % w/v Armeen M2HT in paraffin oil caused slight erythema in 3 and 1 out of 20 treated animals after the 24 and 48-hour readings respectively. No cutaneous reactions were observed in the control group.
-Challenge application of paraffin oil alone caused caused no significant cutaneous reaction.
Executive summary:

The potential of Armeen M2HT to induce delayed contact hypersensitivity was assessed in guinea pigs according to the OECD guideline 406 and conducted in compliance with the principles of Good Laboratory Practice regulations.

The induction phase has been realized both by intradermal route on day 1 (Armeen M2HT 10 % w/v in paraffin oil) and by cutaneous route on day 8 (Armeen M2HT 10% w/v in paraffin oil ) in 2 groups of guinea pigs: 5 males and 5 females for control group and 10 males and 10 females for treated group. The challenge phase was realized on day 22 by cutaneous application of Armeen M2HT 3% and Armeen M2HT 0.5% w/v in paraffin oil on two sites on the right flank (vehicle on the left flank). the cutaneous reactions were scored 24 and 48 hours after the challenge phase.

After the challenge application with 3 % w/v Armeen 2HT in paraffin oil, a slight erythema was observed in 1/ 20 animals of the treated group at the 24 and 48-hour readings. No cutaneous reactions were recorded in the control group.

After the challenge application with 0.5 % w/v Armeen 2HT in paraffin oil, a slight erythema was observed in 3 and 1 out the 20 animals of the treated group at the 24 and 48-hour readings respectively.

The persistent cutaneous reactions observed in 4/20 animals of the treated group after the challenge application may be attributable to delayed contact hypersensitivity but as the cutaneous reactions were more confined to the 24 hour examination, they were more consistent with irritation than delayed contact hypersensitivity.

Therefore, the cutaneous reactions observed in 4/20 animals (20%) of the treated groups were considered as irritative response and the substance was not considered as a skin sensitiser.