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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:
1995-10-23 to 1996-01-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report date:
1996

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes (incl. QA statement)
Type of study:
Buehler test
Justification for non-LLNA method:
The study was conducted before the entry into force of the amendments to Annex VII of REACH. Furthermore, the testing protocol was completed on 18th January 1996. The LLNA according to OECD Guideline 429 was officially adopted by the OECD on the 22nd of July 2010.

Test material

Constituent 1
Chemical structure
Reference substance name:
N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine
EC Number:
219-145-8
EC Name:
N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine
Cas Number:
2372-82-9
Molecular formula:
C18H41N3
IUPAC Name:
N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine
Details on test material:
- Name of test material (as cited in study report): P4150
- Lot/batch No.: PN93-12
- Expiration date of the lot/batch: 21 september 1997
- Storage condition of test material: Room temp in the dark
- Purity: 30.2%
- Physical state: a clear colourless liquid
- Storage condition of test material: room temperature in the dark

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: D. Hall, Newchurch, Staffordshire, England.
- Age at study initiation: ca. 6-7 weeks
- Weight at study initiation: 270 to 315 g
- Housing: in groups of five in suspended metal cages with wire mesh floors
- Diet: A vitamin C enriched guinea-pig diet FD2, ad libitum.
- Water (e.g. ad libitum): Hay was given weekly.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 21
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
for induction: 1% v/v (0.33% test substance) in distilled water

Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
for challenge: 0.5% v/v (0.15% test substance) in distilled water
No. of animals per dose:
20
Details on study design:
RANGE FINDING TESTS: The topical irritancy of a range of dilutions of the test substance was investigated. Based on the results of this study, 1% v/v concentration was chosen for induction and 0.5% v/v concentration was chosen for challenge. The former concentration was the concentration that produced some irritation but did not give rise to adverse effects, while the latter was the maximum concentration not giving rise to irritating effects.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: 6 hours
- Test groups: 20 test animals
- Control group: 10 negative control animals
- Site: left shoulder region, 20 x 20 mm
- Frequency of applications: days 1, 8 and 15
- Duration: 2 weeks
- Concentrations: 1% v/v

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 2 weeks after final induction
- Exposure period: 6 hours
- Test groups: 20 test animals
- Control group: 10 negative control animals
- Site: right flank, 50 x 50 mm
- Concentrations: 0.5% v/v
- Evaluation (hr after challenge): 24 and 48 hours after the removal of the patches
Positive control substance(s):
yes
Remarks:
formalin

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0.15%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Erythema, edema and dryness and sloughing of the epidermis
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0.15%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Erythema, edema and dryness and sloughing of the epidermis
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
not specified
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
Erythema, edema and dryness and sloughing of the epidermis
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
not specified
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
Erythema, edema and dryness and sloughing of the epidermis
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
15%
No. with + reactions:
9
Total no. in group:
10
Clinical observations:
not specified
Remarks on result:
positive indication of skin sensitisation
Remarks:
based on historical positive control data (study period: 25.01.1995 - 25.02.1995)
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
15%
No. with + reactions:
9
Total no. in group:
10
Clinical observations:
not specified
Remarks on result:
positive indication of skin sensitisation
Remarks:
based on historical positive control data (study period: 25.01.1995 - 25.02.1995)

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Conclusions:
The test substance is not sensitising to the skin of guinea-pigs under the conditions of the study. 0.15% of the test material for 6 hours seems to be the threshold for irritation in guinea pigs.
Executive summary:

A contact hypersensitivity study in the guinea pig was performed in compliance with Method B6, 92/69/EEC, in accordance with a modification of the method described in BUEHLER, E.V (1965) Arch. Dermatol., 91, 171. Twenty test and ten control animals were used.

In a preliminary study, the topical irritancy of a range of dilutions of the test substance in distilled water was investigated.

The study identified the concentration suitable for producing irritation suitable for the induction phase and a maximum non-irritant concentration for the challenge:

-For the induction phase; 1% v/v in distilled water. (0.3% a.s.)

-For the challenge phase; 0.5% v/v in distilled water. (0.15% a.s.)

Induction: Prior to each application, the skin on the left shoulder region was clipped free of hair. A 20 x 20 mm patch of surgical gauze (3 layers thick) was saturated with approximately 0.5 mL of the 1% v/v test substance.

The patch was placed on the skin and covered by impermeable plastic adhesive tape (50 mm width “Blenderm”). This was secured with elastic adhesive bandage (50 mm width “Elastoplast”) wound round the torso of the animal and fixed with “Sleek” impervious plastic adhesive tape. Contact was maintained for approximately 6 hours for each induction exposure. Reactions were noted 24 hours after dressing removal.

Induction took place on Days 1, 8, and 15 (a total of 3 inductions).

Control animals were treated similarly to test animals with the exception of test substance omission from the induction applications.

Challenge: Control and test animals were topically challenged two weeks after the final induction. An area of 50 x 50 mm on the right flank of the animal was clipped free of hair. A 20 x 20 mm gauzepatch (3 layers thick) was saturated with approximately 0.5 ml of the 0.5% v/v test substance. The patch was sealed to the skin under a 5cm strip of “Blenderm” covered by “Elastoplast” wound around the trunk and sealed with “Sleek”. After approximately 6 hours the dressings were removed.

Challenge sites were evaluated 24 and 48 hours after removal of the patches. Dermal reactions were scored on a numerical system for oedema and erythema. Evaluations were performed blind (i.e. the scorer had no knowledge of previous scores).

The animals were observed daily for clinical signs of toxicity.

Body weights were recorded on Day 1 (first day of topical application) and on the last day of observations for the challenge. Localised dermal reactions were observed in 3/10 control animals at the 24-hour observation and 1/10 control animals at the 48-hour observation, consisting of varying dermal responses of erythema, oedema and dryness and sloughing of the epidermis. 5/20 test animals at the 24-hour observation and 4/20 test animals at the 48-hour observation exhibited very slight erythema with or without very slight oedema and dryness and sloughing of the epidermis. Two other test animals exhibited dryness and sloughing of the epidermis only, at the 48-hour observation. It was concluded that none of the test animals showed a positive response.