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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:
1984-09-17 to 1984-10-12
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study with acceptable restrictions: No positive control (not required by 1981 version of Test Guideline)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1984
Report date:
1984

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
no
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A skin sensitization test according to OECD 406 has already excisted since 1984 and is sufficient for evaluation of the skin sensitisation potential of the test substance.

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2,4(or 2,4,4)-trimethylhexane-1,6-diamine
EC Number:
247-063-2
EC Name:
2,2,4(or 2,4,4)-trimethylhexane-1,6-diamine
Cas Number:
25513-64-8
Molecular formula:
C9H22N2
IUPAC Name:
2,2,4-trimethylhexane-1,6-diamine; 2,4,4-trimethylhexane-1,6-diamine
Test material form:
other: liquid
Details on test material:
2,2,4(or 2,4,4)-trimethylhexamethylenediamine of Hüls AG, purity > 99.7 %

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS: 
- Strain: Dunkin-Hartley (Bor: DHPW)
- Sex: female
- Source: Winkelmann, Borchen (Germany)
- Weight at study initiation: 428 g (mean)
- Controls: vehicle treatment
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 1 °C
- Humidity (%): 60 +/- 5 %
- Air changes (per hr): 15 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
1st application: Induction 0.1 % intracutaneous
2nd application: Induction 10 % occlusive epicutaneous
3rd application: Challenge 2 % occlusive epicutaneous
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
1st application: Induction 0.1 % intracutaneous
2nd application: Induction 10 % occlusive epicutaneous
3rd application: Challenge 2 % occlusive epicutaneous
No. of animals per dose:
20 females (test) / 20 females (control)
Details on study design:
ADMINISTRATION/EXPOSURE 
- Induction schedule: injection followed 1 week later by patch treatment  for 48 hours
- Injection details: 0.1 ml each at 6 positions on shoulders:   
2 x Freund's Complete Adjuvant   
2 x test substance 0.1 % in aqua dest.   
2 x Freund's Complete Adjuvant / 0.2 % test substance (50:50)   
simultaneous and symmetrical application of each solution   
controls: aqua dest. instead of test substance
- Challenge schedule: 2 weeks after end of induction patch treatment for  24 hours
- Concentrations used for challenge: 2 %; readings 24, 48, and 72 hours  after removal of patch
- Rechallenge: no
- Positive control: no
EXAMINATIONS
- Grading system: possible scores 0 / 1 / 2 / 3       
0 % of animals scored > 0: no sensitization  
1 -  8 % of animals scored > 0: very slight sensitization  
9 - 28 % of animals scored > 0: slight sensitization
29 - 64 % of animals scored > 0: distinct sensitization
65 - 80 % of animals scored > 0: severe sensitization
81 -100 % of animals scored > 0: extreme sensitization
Challenge controls:
similarly treatment, but with vehicle alone
Positive control substance(s):
no

Results and discussion

Positive control results:
not examined

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
2 %
No. with + reactions:
16
Total no. in group:
20
Clinical observations:
see "Remarks on results"
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 2 %. No with. + reactions: 16.0. Total no. in groups: 20.0. Clinical observations: see "Remarks on results".
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
Vehicle
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no reactions
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Vehicle. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no reactions.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
2 %
No. with + reactions:
16
Total no. in group:
20
Clinical observations:
see "Remarks on results"
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 2 %. No with. + reactions: 16.0. Total no. in groups: 20.0. Clinical observations: see "Remarks on results".
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Vehicle
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no reactions
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: Vehicle. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no reactions.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test chemical
Dose level:
2 %
No. with + reactions:
8
Total no. in group:
20
Clinical observations:
see "Remarks on results"
Remarks on result:
other: Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 2 %. No with. + reactions: 8.0. Total no. in groups: 20.0. Clinical observations: see "Remarks on results".
Reading:
other: 3 rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
Vehicle
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no reactions
Remarks on result:
other: Reading: other: 3 rd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: Vehicle. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no reactions.

Any other information on results incl. tables

RESULTS OF TEST
- Sensitization reaction: 
  16/20 animals positive at 24 and 48 hours = severe sensitization
  8/20 animals positive at 72 hours
  no animals positive in control group
- Clinical signs: 
  1st induction, FCA application sites: distinct erythema and edema and slight necrosis
  1st induction, FCA + test substance application sites: severe erythema and edema, necroses and brown / black discoloration
  1st induction, vehicle application sites: slight erythema
  2nd induction, test group: inflammation, black discoloration
  2nd induction, control group: slight inflammation at few injection sites

Applicant's summary and conclusion

Conclusions:
In a guinea pig maximization test (according to OECD Guideline 406, positive controls not required by 1981 guideline version), sensitization was observed in 16 of 20 female animals. Based on these results, 2,2,4(or 2,4,4)-trimethylhexane-1,6 -diamine should be classified as contact sensitizer.
Executive summary:

The skin sensitizing properties of 2,2,4(or 2,4,4)-trimethylhexane-1,6-diamine were conducted in a guinea pig maximization test according to OECD 406 (positive controls not required by 1981 guideline version). Twenty female guinea pigs were intradermally injected with a 0.1 % concentration of 2,2,4(or2,4,4)-trimethylhexane-1,6 -diamine (in water) and one week later epidermally exposed to a 10 % concentration of test substance. Twenty control animals were similary treated, but with vehicle alone. Two weeks after the epidermal application all animals were challegend with 2 % test substance and the vehicle.

Under the conditions of the test sensitizing effects were observed in 16 of the 20 animals at 24 and 48 hours. Based on these results, 2,2,4(or 2,4,4)-trimethylhexane-1,6 -diamine should be classified as contact sensitizer.