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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
Study period:
Aug 1992 - december 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report date:
1993

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
no test substance purity given
GLP compliance:
yes (incl. QA statement)
Type of study:
Buehler test
Justification for non-LLNA method:
Test was performed before LLNA method has to be used as first choice.

Test material

Constituent 1
Chemical structure
Reference substance name:
Amines, polyethylenepoly-, triethylenetetramine fraction
EC Number:
292-588-2
EC Name:
Amines, polyethylenepoly-, triethylenetetramine fraction
Cas Number:
90640-67-8
Molecular formula:
C6H18N4, C8H20N4
IUPAC Name:
(2-aminoethyl)({2-[(2-aminoethyl)amino]ethyl})amine; tris(2-aminoethyl)amine
Test material form:
liquid

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Buckberg Lab Animals, Tomkins Cove, New York
- Age at study initiation: 4 - 6 weeks
- Weight at study initiation: 301-369g
- Housing: individually in stainless steel wire mesh cages
- Diet (e.g. ad libitum): purina guinea pig diet, ad libitum
- Water (e.g. ad libitum): fresh tap water, ad libitum
- Acclimation period: minimum 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light


IN-LIFE DATES: From: 5 Dec 1992 To: 12 Dec 1992 (Dose-Range-Finding)
IN-LIFE DATES: From: 14 Dec 1992 To: 13 Jan 1993 (Assay)

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, occlusive
Vehicle:
other: 80% ethanol
Concentration / amount:
0.3 mL/site (70%)
Day(s)/duration:
3 inductions, one week in between, 6 h exposure duration
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
0.3 mL/site (50%)
Day(s)/duration:
1 and 2
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
dose range finding: 12 (6 males, 6 females)
test article: 20 (10 males, 10 females)
positive control: 5 (3 males, 2 females)
negative control: 10 (5 males, 5 females)
Details on study design:
RANGE FINDING TESTS:
2 males, 2 females: each animal is exposed to 4 different concentrations: 1%, 10%, 50%, 100%; 80% ethanol as the vehicle.
1 male, 1 female: each animal is exposed to 4 different concentrations: 60%, 70%, 80%, 90%; 80% ethanol as the vehicle.
2 males, 2 females: each animal is exposed to 4 different concentrations: 1%, 10%, 50%, 90%; acetone as the vehicle
1 male, 1 female: each animal is exposed to 4 different concentrations: 20%, 30%, 40%, 50%; acetone as the vehicle
primary challenge responses were graded
Highest non-irritatting concentration = concentration that induced responses not exceeding 2 + and 2 0 grades in the group of 4 animals.
the dose chosen for induction: 70%
the dose chosen for challenge: 50%


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 4 (3 inductions, 1 challenge)
- Exposure period: -
- Test groups: test substance in vehicle (80% ethanol)
- Control group: vehicle only (80% ethanol)
- Site: L shoulder (1st and 2nd induction), R shoulder (3rd induction)
- Frequency of applications: once a week
- Duration: 6 h
- Concentrations: 70%


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: day 29
- Exposure period: -
- Test groups: test substance in vehicle (aceton)
- Control group: vehicle (aceton) only (left flank), test article (right flank)
- Site: naive site on left flank
- Concentrations: 50%
- Evaluation (hr after challenge): 24 and 48h

Other:
24h after challenge and rechallenge, all animals were depilated with Neet Cream Hair Remover (Whitehall Laboratories, Inc., New York). A minimum of 2h after depilation test sites were graded. The grading was repeated 24h later (48h grade).
All animals were euthanized by CO2 inhalation at study termination.
Challenge controls:
The negative control group was challenged with vehicle (aceton) on the left flank and test article on the right flank.
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitrobenzene

Results and discussion

Positive control results:
Sensitising effects observed in all 5 animals of the positive control group.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
70% induction, 50% challenge
No. with + reactions:
18
Total no. in group:
20
Clinical observations:
severity= 2.2
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
70% induction, 50% challenge
No. with + reactions:
19
Total no. in group:
20
Clinical observations:
severity= 2.4
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50% challenge
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
severity= 0.0
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50% challenge
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
severity= 0.0
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.3% challenge
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
severity= 3.0
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.3% challenge
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
severity= 3.0
Remarks on result:
positive indication of skin sensitisation

Any other information on results incl. tables

The results above on the test group have been corrected, based on the raw data presented in the study.

Applicant's summary and conclusion

Interpretation of results:
other: CLP/EU GHS Category 1B (H317) according to Regulation (EC) No 1272/2008
Conclusions:
Based upon the observations made in the assay, the test article (induction and challenge concentrations 70% and 50%, respectively), caused delayed contact hypersensitivity in guinea pigs.
Executive summary:

A Buehler test for skin sensitisation with the test substance was carried out in guinea pigs. Based on preliminary testing, the concentration chosen for induction was 70% and the concentration chosen for challenge 50% (occlusive applications). At the first reading (24 h after challenge), 18/20 animals showed skin reactions; at the second reading (48 h after challenge), 19/20 animals were positive. It was concluded that the test substance is a skin sensitiser.