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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1982
Report Date:
1982
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
(1981-05-12)
Deviations:
yes
Remarks:
(number of animals in test and control group smaller than recommendations in guideline (12 vs. 20), no data of positive controls given)
Principles of method if other than guideline:
Test procedure was conducted according to Magnusson and Kligmann (Magnusson, B. and Kligman A.M.: The Identification of Contact Allergens by Animal Assay. The Guinea Pig Maximization Test. J. Invest. Dermatol. 52, 268 – 276 (1969))
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Equivalent study of good quality from 1982 is available, additional conduct of LLNA is not deemed scientifically justified.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): N,N,N',N'-Tetramethyl-1,6-hexanediamine
- Analytical purity: 97.5 %
- Expiration date of the lot/batch: 1984-01 (on account of the sufficient stability no analytical evidence of the stability was necessary during the study)
- Impurities (identity and concentrations): N,N',N'-Trimethyl-1,6-hexanediamine 2%

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Lippische Versuchstierzucht, Hagemann GmbH & Co. KG, Extertal, Germany
- Weight at study initiation: 235- 310 g
- Housing: 6 animals per cage (Type IV Makrolon cage)
- Diet: SSNIFF GK 4mm, standard diet for rabbits and guinea pigs, Ssniff Versuchstierdiäten GmbH, Soest, Germany
- Water (e.g. ad libitum): ad libitum (fully demineralized from monday to friday, tap water on saturdays and sundays; about 2g ascorbic acid/ 10 L water was added to the drinking water twice per week)
- Acclimation period: at least 7 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 2
- Humidity (%): 30- 70
- Air changes (per hr): 25- 20
- Photoperiod (hrs dark / hrs light): 12/ 12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
pretest: 0.5, 1, 3, 5, 10 % aquous solution
Induction: 5% aquous solution (for injection and epicutaneous application)
Challenge: 3% aquous solution
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
pretest: 0.5, 1, 3, 5, 10 % aquous solution
Induction: 5% aquous solution (for injection and epicutaneous application)
Challenge: 3% aquous solution
No. of animals per dose:
6 (control group 1 and 2)
12 (test group)
Details on study design:
RANGE FINDING TESTS:
Amount applied: 0.15 g of test substance preparation occlusive (by filter strips soaked with a 0.5mm layer of the test substance)
Preparations of the test substance: 0.5, 1, 3, 5, 10 % in aqua dest
Exposure period: 24 hrs
Number of animals: 4/ test concentration
Readings: 24 and 48 hrs after application

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous)
- Exposure period: single injection (intradermal), after 7 days epicutaneous application (48 hrs)

Intradermal iinjection:
- Site: shoulder region
- Frequency of application: once
- Test groups:
3 pairs of injections
1.) 1:1 mixture (Freund's adjuvant/ water)
2.) test substance preparation (5%)
3.) mixture of Freund's adjuvant/water (1:1) and test substance preparation (5%)

- Control group (2 identical control groups)
3 pairs of injections
1.) 1:1 mixture (Freund's adjuvant/ water)
2.) aqua dest
3.) mixture of Freund's adjuvant/water (1:1) and aqua dest

Epicutaneous application: test substance 5% in aqua dest. (controls were not applied), occlusiv

- Site: shoulder region (same region as with intradermal injection)
- Frequency of applications: once, 7 days after intradermal injection
- Duration: 48 hrs
- Concentrations: 5% test substance in aqua dest. (0.3 g test substance preparation using 2x4 cm test filter paper strip)

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge:
1.) First challenge:
14 days after epicutanous application
2.) Rechallenge:
about one week after first challenge

- Exposure period: 24 hrs for challenge and rechallenge
- Test groups: test substance in aqua dest.
- Control group: 1st control group: test substance in aqua dest (3%), 2 nd control group: aqua dest
- Site: right flank (1st challenge), left flank(2nd challenge)
- Concentrations: 3%
- Evaluation (hr after challenge): 24, 48, 72

Assessment of Skin findings:
1. Erythema and eschar formation:
a) No erythema: 0
b) Very slight erythema (barely perceptible): 1
c) Well-defined erythema: 2
d) Moderate to severe erythema: 3
e) Severe erythema (beet redness) to slight eschar
formation (injuries in depth): 4
2. Edema formation:
a) No edema: 0
b) Very slight edema (barely perceptible): 1
c) Slight edema (edges of area weil defined by definite raising): 2
d) Moderate edema (raised approximately 1 mm): 3
e) Severe edema (raised more than 1 mm and extending beyond the area of exposure: 4

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
3 % (first control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no observations
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 3 % (first control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no observations.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
3 % (first control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no observations
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 3 % (first control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no observations.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle only (second control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no application since distilled water was used as vehicle
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: vehicle only (second control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no application since distilled water was used as vehicle.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle only (second control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no application since distilled water was used as vehicle
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: vehicle only (second control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no application since distilled water was used as vehicle.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
3%
No. with + reactions:
0
Total no. in group:
12
Clinical observations:
no observations
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 3%. No with. + reactions: 0.0. Total no. in groups: 12.0. Clinical observations: no observations.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
12
Clinical observations:
no observations
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 12.0. Clinical observations: no observations.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
3 % (first control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no observations
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 3 % (first control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no observations.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
3 % (first control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no observations
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 3 % (first control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no observations.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
3 % (second control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no observations
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 3 % (second control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no observations.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
3 % (second control group)
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
no observations
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 3 % (second control group). No with. + reactions: 0.0. Total no. in groups: 6.0. Clinical observations: no observations.
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
12
Clinical observations:
no observations
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 12.0. Clinical observations: no observations.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
12
Clinical observations:
no observations
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 12.0. Clinical observations: no observations.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Executive summary:

A test on substance N,N,N',N'-Tetramethyl-1,6-hexanediamine was conducted for assessing the skin sensitization potential of the compound. The test was conducted according to the method of Magnusson and Kligmann which was comparable to the OECD guideline study 406 (1981) with deviations (no data of positive control, smaller number of test and control animals; 12 vs. 20).

The substance was applied on female guinea pigs with 2 groups of negative control (6 animals per group) and one test group (12 animals). For induction, a 5% solution of test substance with water was injected intradermally and after 7 days applied epicutaneously. Then a first challenge 14 d after percutanous application and a 2nd challenge one week later was done with control and test substance (3% in aqua dest.). No positive results were obtained for the test substance -neither in the challenge nor in the rechallenge application. Consequently the substance is to be considered as non-sensitizing in the Magnusson-Kligman-Test under the test conditions chosen.