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Description of key information

In a Buehler test according to OECD Guideline 406, the test substance was demonstrated to be sensitising to the skin (Armondi, 1991).

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 January 1991 - 7 March 1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
The Murine Local Lymph Node Assay (LLNA) is the first-choice method for in vivo testing according to the REACH Regulation. However, this test was performed before entry into force of the REACH Regulation.
Specific details on test material used for the study:
- Name of test material (as cited in study report): 6398-24-1, Project #90-018
- Substance type: clear colorless liquid
- Physical state: liquid
- Analytical purity: responsibility of the sponsor
- Stability under test conditions: no apparent change in the physical appearance of the test article during administration
- Storage condition of test material: responsibility of the sponsor
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
- Weight at study initiation: 300-700 g
- Age at study initiation: no data
- Housing: individually in 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): 20 ± 3°C
- Humidity (%): 30-70 %
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light

Route:
epicutaneous, occlusive
Vehicle:
other: 80% ethanol (inductions) and acetone (challenge)
Concentration / amount:
Volume administered: 0.3 ml/dose
Induction: 5.0% (v/v); challenge: 5.0% (v/v)
Route:
epicutaneous, occlusive
Vehicle:
other: 80% ethanol (inductions) and acetone (challenge)
Concentration / amount:
Volume administered: 0.3 ml/dose
Induction: 5.0% (v/v); challenge: 5.0% (v/v)
No. of animals per dose:
dose range: 8 (4 males, 4 females);
test article: 20 (10 males, 10 females);
positive control: 5 (2 males, 3 females);
negative control: 10 (5 males, 5 females)
Details on study design:
RANGE FINDING TESTS:
8 animals (4 males, 4 females) were each exposed to 4 concentrations of the test material in either 80% ethanol (test concentrations of 1, 10, 50 and 100%) or acetone (test concentrations of 1.0, 2.5, 5.0 and 10%)
Primary irritation responses were graded after 24h as follows:
0 = no reaction
+ = slighly patchy erythema
1 = slight of confluent or moderate patchy erythema
2 = moderate erythema
3 = severe erythema with/without edema
The highest non-irritatting concentration was that concentration in the vehicle that induced responses not exceeding two + and two 0 grades in the group of 4 animals.
Based upon the results of the dose-range-finding studies and in discussion with the sponsor, the test article was dosed at 5.0% in 80% ethanol for induction and at 5.0% in acetone for challenge.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 inductions
- Exposure period: Once a week for three weeks, a total of three six-hour inductions.
- Test groups: test substance in vehicle (80% ethanol)
- Control group: vehicle only
- Site: left shoulder
- Frequency of applications: once a week
- Duration: 6 h
- Concentrations: 5.0%


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 14 days after the last induction sensitizing exposure
- Test groups: test substance in vehicle (acetone)
- Control group: vehicle only (left flank), test article (right flank)
- Site: naive site on left side
- Concentrations: 5.0%
- Evaluation (hr after challenge): 24 and 48h


OTHER:
24h after challenge, all animals were depilated with Neet Cream Hair Remover (Whitehall Laboratories, Inc., New York). A minimum of 2h after depilation, test sites were graded.
Challenge controls:
The negative control group was challenged with vehicle (acetone) on the left flank.
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitrobenzene (0.3%)
Positive control results:
A positive response was elicited in the animals receiving the positive control article, 1-chloro-2,4-dinitrobenezen (DNCB)
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5.0%
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
severity= 0.5
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5.0%
No. with + reactions:
6
Total no. in group:
20
Clinical observations:
severity= 0.7
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.3%
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
severity= 3.0
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.3%
No. with + reactions:
5
Total no. in group:
5
Clinical observations:
severity= 3.0
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle (acetone)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
severity= 0.0
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle (acetone)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
severity= 0.0
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
other: naive control
Dose level:
5.0% test article
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
severity= 0.1
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
other: naive control
Dose level:
5.0% test article
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
severity= 0.1

Results dose-range finding test:

Vehicle 80% ethanol: 1.0% dose caused no reaction, 10% dose caused slightly patchy erythema (1/4), slight or confluent or moderate patchy erythema (2/4) and severe erythema (1/4), 50% and 100% doses caused severe erythema.

Vehicle acetone: 1.0% and 2.5% doses caused no reaction, 5.0% dose caused no reaction (1/4) and slightly patchy erythema (3/4), 10% dose caused no reaction (2/4), slightly patchy erythema (1/4) and severe erythema (1/4).

The highest non-irritating dose (max. two animals with slightly patchy erythema) for challenge exposure was chosen to be 5% in acetone. The highest dose to cause mild irritation for induction exposure was chosen to be 5% in 80% ethanol.

Interpretation of results:
Category 1B (indication of skin sensitising potential) based on GHS criteria
Conclusions:
Based upon the observations made in the assay, the substance induced and challenged at a 5.0% concentration caused delayed contact hypersensitivity in guinea pigs.
Endpoint:
skin sensitisation: in vitro
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Specific details on test material used for the study:
- Name of test material (as cited in study report): Methoxypropylamine
- Batch: FABRICATION F 6612 r (860208)
- Purity c.a. 99%
- Supplier: Atochem, La Chambre
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Iffa Credo : Les Oncins- 69210 L'Arbresle -France
- Age at study initiation:
- Weight at study initiation: 300 ta 500 g.
- Housing: housing by groups of 5 (or of 2 for the preliminary studies), in polystyrene cages,
- Diet: Guinea-pig complete pelleted maintenance food, ad libitum
- Water : Softened and filtered drinking water (15 p.m), ad libitum
- Acclimation period: minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 to 70
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
1st application: Induction 0.25 % intracutaneous
2nd application: Induction 2.5 % occlusive epicutaneous
3rd application: Challenge 0.5 % occlusive epicutaneous
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
1st application: Induction 0.25 % intracutaneous
2nd application: Induction 2.5 % occlusive epicutaneous
3rd application: Challenge 0.5 % occlusive epicutaneous
No. of animals per dose:
20 test, 10 control
Positive control substance(s):
no
Key result
Reading:
other: Reading 6, 24 or 48h
Group:
test group
Dose level:
0.5%
No. with + reactions:
2
Total no. in group:
20
Key result
Reading:
other: Reading 6, 24 or 48h
Group:
negative control
Dose level:
0.5%
No. with + reactions:
0
Total no. in group:
10
Interpretation of results:
GHS criteria not met
Conclusions:
From the macroscopic and histological results obtained under the experimental conditions, the test article provoked a slight cutaneous sensitizing reaction in 2 out of the 20 treated guinea-pigs (10 %); no cutaneous abnormalitv was noted in the 10 control guinea-pigs.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

One key and one supporting study were available.

Armondi (1991) investigated the sensitising properties of the test substance in a Buehler test with male and female Hartley guinea pigs. The study examined the skin sensitising effect on guinea pigs, using 20 animals in total. Positive, solvent and negative controls were included in the study. Animals were induced and challenged with 5.0% (v/v) test substance. Based upon the observations made in the assay, the substance caused delayed contact hypersensitivity in guinea pigs.

Guillot (1986) concluded that the substance should not be classified as sensitising. From the macroscopic and histological results obtained under the experimental conditions, the test article provoked a slight cutaneous sensitizing reaction in 2 out of the 20 treated guinea-pigs (10 %); no cutaneous abnormalitv was noted in the 10 control guinea-pigs.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the study results and the criteria of the CLP Regulation, the test substance is classified as skin sensitising substance (category 1B).