Registration Dossier

Administrative data

Description of key information

- GPMT, Not sensitising (OECD 406, GLP, Rel.1)
- Not sensitising in humans up to 10% (1 HRIPT).

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:
From December 22, 1997 to January 23, 1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
details of environmental conditions not reported
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
At the time of study completion (1998), the LLNA OECD test method was not adopted.
Species:
guinea pig
Strain:
Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles RIVER, 76410 Saint-Aubin-Les-Elbeuf, France.
- Weight at study initiation: 315-374 g
- Housing: Animals were housed individually in polypropylene cage (310 x 465 x 190).
- Diet: Complete pelleted diet UAR 106 (91360 - Epinay sur Orge, France), ad libitum
- Acclimation period: 5 days

IN-LIFE DATES: From December 22, 1997 to January 23, 1998
Route:
intradermal
Vehicle:
other: mineral oil
Concentration / amount:
40%
Day(s)/duration:
Day1
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: acetone
Concentration / amount:
80%
Day(s)/duration:
Day 8
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:
2.5 and 5%
Day(s)/duration:
Day 21
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
5 and 10 animals for control and treatment group, respectively
Details on study design:
RANGE FINDING TESTS:
- Determination of the MIC (Maximal concentration slightly to moderately irritating) by intradermal route (MICID): One week before the main test, two guinea-pigs received intradermal injections of test material at 10, 20, 40 and 80 % w/v in mineral oil and evaluated for skin reactions at 24 h after injections. The concentration selected for the main study (intradermal induction) was 40 % w/v in mineral oil.
- Determination of the MIC by topical route (MICT): Between Day -8 and Day 4, two guinea pigs were applied topically with test material at 10, 20, 40 and 80 % in acetone and acetone only under occlusive patch for 48 ± 1 h and cutaneous irritation reactions were evaluated after the patch removal. The concentration selected for the main study (topical induction) was 80 % in acetone.
As irritation reactions were still observed with the lowest concentration (10 %), in order to determine the concentrations to be used for the maximal non irritant concentration (MNIC) determination, a second test was performed in two other animals with lower concentrations (1.25, 2.5, 5 and 10 % in acetone).
-Determination of the MNIC by topical route: One week before the main test, at Day -8, two guinea pigs received the intradermal injection of 0.1 mL of the 3 following preparations on both sides of the spinal column, on the clipped dorsal area: Freund's Complete Adjuvant (FCA) diluted at 50 % with distilled water (v/v); vehicle : mineral oil; a 50/50 mixture of the FCA diluted at 50 % with distilled water (v/v) and mineral oil. After a 21 day resting period, the animals were applied topically with test material at 2.5 and 5% in acetone (determined in w/v from the results obtained during the evaluation of the MICT) under occlusive patches for 24 ± 1 h. Skin reactions were evaluated 24, 48 and 72 h after patch removal. The concentrations selected for the main study (challenge phase) were 2.5 and 5 % in acetone (MNIC/2 and MNIC).

MAIN STUDY
A. INDUCTION EXPOSURE: INTRADERMAL
- No. of exposures: One
- Test groups: Intradermally injected with 3 injections (0.1 mL each) of FCA and distilled water in the ratio 1:1 (v/v); a 40 % w/v formulation of the test material in mineral oil; a 40 % w/v formulation of the test material in a 1:1 (v/v) preparation of FCA and distilled water, on Day 1.
- Control group: Intradermally injected with 3 injections (0.1 mL each) of FCA and distilled water in the ratio 1:1 (v/v); vehicle (mineral oil); a mixture 1:1 (v/v) of FCA plus distilled water and mineral oil, on Day 0.
- Site: On both sides of the spinal column, at the scapular level
- The injections 1 and 2 were performed close to each other, as nearer as possible to the head. The injection 3 was performed towards the caudal part of the experimental area.

On Day 7: After 6 days resting period, animals were clipped at the injection site level. The experimental area of the control and treated group animals were coated with 0.5 mL of a sodium lauryl sulphate at 10 % in petrolatum to produce a local irritation.

B. INDUCTION EXPOSURE: TOPICAL
- No. of exposures: One, on Day 8
- Exposure period: 48 ± 1 h
- Test groups: 0.5 mL of the test material at 80 % in acetone was applied to the filter paper patch of 8 cm2 (2x4 cm), put on a piece of gauze of 8 cm2 (2x4 cm) put on a square Blenderm occlusive film of 25 cm2. The occlusive patch was held in place with an adhesive Micropore film put around the animal trunk during 48 ± 1 h exposure.
- Control group: 0.5 mL of the vehicle (acetone COOPER batch 122470) was applied according to the method described above and let in contact with the skin during 48 ± 1 h exposure.
- Site: Same intradermally injected area
- Frequency of applications: Single application

C. CHALLENGE EXPOSURE: TOPICAL
- No. of exposures: One
- Day of challenge: Day 21
-After a 11-day rest period, the guinea-pigs from both groups were clipped one last time at the dorso-lumbar region, the skin surface chosen corresponding to a zone which had not previously been in contact with the test product.
- Exposure period: 24 ± 1 h
- Test groups and control groups: 0.2 mL of the test material at 2.5 and 5 % in acetone were applied respectively on 2 patches made with a square of filter paper of about 2 cm2 which receives the product, put on a square of gauze of 4 cm2, put in the middle of an occlusive Blenderm film of 25 cm2. The patches were loosely held on the right flank of each animal by a Micropore film. As the vehicle used was not distilled water, 0.2 mL of acetone was put on a patch and applied to the same flank of each animal as the test material.
- Evaluation (h after removal of challenge patch): 24, 48 and 72 h

OTHER:
- Body weight of each animal was recorded at the start and end of the study.
Challenge controls:
See "Details on study design"
Positive control substance(s):
yes
Remarks:
2-Mercapto Benzothiazole
Positive control results:
The sensitivity and the validity of the experimental technique adopted were confirmed by the study EVIC-CEBA Td 718 of October 09, 1997. The product 2-Mercapto Benzothiazole appeared highly to very highly sensitizing for the guinea-pig (80 % to 90 % of reactive animals at the 24, 48 and 72 h reading - class 4) when it was used at challenge diluted at 5 % and 2 % with petrolatum.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5 % in acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5 % in acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
5 % in acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
2.5 % in acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
2.5 % in acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
2.5 % in acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
other: Solvent control
Dose level:
Acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Solvent control
Dose level:
Acetone
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
other: Solvent control
Dose level:
Acetone
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
2.5 or 5 % in acetone or acetone only
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
2.5 or 5 % in acetone or acetone only
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
2.5 or 5 % in acetone or acetone only
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
5 to 2%
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Remarks:
80 to 90% of reactive animals; exact number not reported
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
5 to 2%
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Remarks:
80 to 90% of reactive animals; exact number not reported
Reading:
other: 3rd readling
Hours after challenge:
72
Group:
positive control
Dose level:
5 to 2%
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Remarks:
80 to 90% of reactive animals; exact number not reported

Determination of the sensitizing potential (main test):

In the control group, no local irritation reaction was observed 24, 48 and 72 h after patch removal.

In the treated group (previously induced by intradermal and epidermal routes on another site of the skin), the presence of slight local irritation reaction in 1 animal for the three reading times after application of the tested substance at the concentrations 2.5 and 5 % in acetone. The same animal presented a slight erythematous reaction after application of the vehicle only for the 24 and 48 h readings.

 

Table 7.4.1/1: Main study – Challenge phase results

 

Group / Animal No.

5 % with acetone

2.5 % with acetone

Acetone

24 h

48 h

72 h

24 h

48 h

72 h

24 h

48 h

72 h

Group 1: Control group

8983

0

0

0

0

0

0

0

0

0

8984

0

0

0

0

0

0

0

0

0

8985

0

0

0

0

0

0

0

0

0

8986

0

0

0

0

0

0

0

0

0

8987

0

0

0

0

0

0

0

0

0

Reactive animals (%)

0

0

0

0

0

0

0

0

0

Group 2: Treated group

8988

1

2

1

1

1

1

1

1

0

8989

0

0

0

0

0

0

0

0

0

8990

0

0

0

0

0

0

0

0

0

8991

0

0

0

0

0

0

0

0

0

8992

0

0

0

0

0

0

0

0

0

8993

0

0

0

0

0

0

0

0

0

8994

0

0

0

0

0

0

0

0

0

8995

0

0

0

0

0

0

0

0

0

8996

0

0

0

0

0

0

0

0

0

8997

0

0

0

0

0

0

0

0

0

Reactive animals (%)

10

10

10

10

10

10

10

10

0

 

General state - Mortality

The general state of the guinea-pigs was not modified throughout the test period.

The weight growth of all the animals was satisfactory. The mean weight gain calculated for the treated animals during the test period was not significantly different from the control animals.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the test item is not classified as sensitiser according to the criteria of Annex VI to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In a Magnusson & Kligman maximisation study (GPMT) performed according to OECD Guideline 406 and in compliance with GLP, 10 female Hartley guinea pigs were induced with three pairs of intradermal injections (0.1 mL each) of FCA and distilled water in the ratio 1:1 (v/v); a 40 % w/v formulation of the test material in mineral oil; a 40 % w/v formulation of the test material in a 1:1 (v/v) preparation of FCA and distilled water, on Day 1 on both sides of the spinal column, at the scapular level. Whereas, control group of 10 females was intradermally induced with 0.1 mL of FCA and distilled water in the ratio 1:1 (v/v); vehicle (mineral oil); a mixture 1:1 (v/v) of FCA plus distilled water and mineral oil. On Day 8, the experimental area of the control and treated group animals were coated with 0.5 mL of a sodium lauryl sulphate at 10 % in petrolatum to produce a local irritation. On Day 8, the same area was topically induced with 80 % test material in acetone via occluded filter paper patch for 48 ± 1 h in test group, whereas control group was applied with vehicle alone. On Day 21, a challenge filter paper patch loaded with test material (2.5 and 5 % in acetone) was applied to the right flank of all animals, and acetone was applied to the same flank of each animal as the test material. The test concentrations for the main study were determined from a sighting study.

In the control group, no local irritation reaction was observed after 24, 48 and 72 h after patch removal. In the treated group, the presence of slight local irritation reaction in 1 animal for the three reading times after application of the tested substance at the concentrations 2.5 and 5 % in acetone. The same animal presented a slight erythematous reaction after application of the vehicle only for the 24 and 48 h readings. The sensitivity and validity of the experiment was confirmed with known sensitiser, 2-Mercapto benzothiazole.

 

Under the test conditions, the test material is not classified according to the annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for sensitisation endpoint.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

A key study was identified (EVIC-CEBA, 1998, rel. 1). In a Guinea-pig maximisation test (GPMT) performed according to OECD guideline No. 406 and in compliance with GLP, the test material diluted in mineral oil at 40% was administered by injection for intradermal induction. On Day 8, the experimental area of the control and treated group animals were coated with 0.5 mL of a sodium lauryl sulphate at 10 % in petrolatum to produce a local irritation. Topical induction was performed with the test material diluted in acetone at 80 % on Day 8. For the challenge, the test material was applied topically at 5 and 2.5 % in acetone. The test concentrations for the main study were determined from a sighting study. The sensitivity and validity of the experiment was confirmed with known sensitiser, 2-Mercapto benzothiazole.

In the control group, no local irritation reaction was observed after 24, 48 and 72 h after patch removal. In the treated group, the presence of slight local irritation reaction in 1 animal for the three reading times after application of the tested substance at the concentrations 2.5 and 5 % in acetone. The same animal presented a slight erythematous reaction after application of the vehicle only for the 24 and 48 h readings. Under the test conditions, the test material should be considered as not sensitising by contact with the skin.

A human repeated insult patch test has also been performed with the substance at up to 10 % (TKL, 1998).Under the conditions employed in this study, there was no evidence of sensitisation to the test material at 10 %.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.

Self classification:

Based on the available data, no additional self-classification is proposed according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

No data was available for respiratory sensitisation. However, this substance is not a skin sensitizer, therefore according to Figure R.7.3 -2 of the Chapter R.7 (V 4.1 - October 2015) the chemical is not considered as a respiratory sensitizer.