Registration Dossier

Administrative data

Description of key information

Skin: (Buehler, GPMT, VAA): positive

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
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
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.
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)
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
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
Positive control results:
Sensitising effects observed in all 5 animals of the positive control group.
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

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

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.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

A Buehler test equivalent to OECD guideline 406 with Amines, polyethylenepoly-, triethylenetetramine fraction was carried out in guinea pigs (Armondi, 1992). 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. Positive control with 1-chloro-2,4-dinitrobenzene resulted in 100% positive reactions showing the test system is valid. Based on the results a classification in CLP/EU GHS Category 1B is warranted.

A supporting GPMT study performed equivalent to OECD guideline 406 with Amines, polyethylenepoly-, triethylenetetramine fraction in guinea pigs is available showing similar results (Auletta, 1990). After a range finding study the concentration used in the study were set at 95% for the induction and 50% for the challenge via topical exposure and 5.0% for the intradermal exposure. One of the test animals died but the death was not related to the test substance. Eleven of the nineteen animals challenged exhibited clear dermal responses (scores of 1 or higher) 24 and/or 48 hours after challenge; four and seven additional animals exhibited scores of 0.5 at 24 or 48 hour challenge reading, respectively. According to the grading scale in the OECD guideline 406 only scores from 1 to 3 are relevant. Therefore, a 58% positive response to the test substance was observed after 24 and 48 hours, respectively. Thus, a classification in CLP/EU GHS Category 1B is warranted.

Four further supporting skin sensitisation studies with reliability 4 are available. All those studies are shortly described in publications. Three GPMT studies (Maisey, 1988; Leung, 1997; Thorgeirsson, 1978) and one mouse ear swelling test (Maisey, 1988) were described. All studies describe a skin sensitising potential for Amines, polyethylenepoly-, triethylenetetramine fraction: In the GMPT study from Maisey 90% guinea pigs showed positive response after 0.5% test substance treatment at intradermal induction. 74% guinea pigs showed positive response after 5% test substance treatment at intradermal induction (Leung 1997) and 80% guinea pigs showed positive response after 0.5% test substance treatment at intradermal induction (Thorgeirsson, 1978).

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Reliable data Amines, polyethylenepoly-, triethylenetetramine fraction on skin sensitisation concludes that the registered substance meets the criteria for classification according to Regulation (EC) No. 1272/2008. Taking into account all data from animal and human studies no sub-categorisation is possible. The test substance is therefore classified as sensitising to skin Cat. 1 (H317).