[3aR-(3aα,5aβ,9aα,9bβ)]-dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan

Endpoint summary

Description of key information

- GPMT, Not sensitising (OECD 406, GLP, WoE, Rel.2)
- LLNA, Not sensitising (Read-Across, OECD 429, GLP, WoE, Rel. 1)
- OET, Not sensitising (OECD 406, GLP, S, Rel.3)
- Buehler, Not sensitising (S, Rel.3)
- Not sensitising in humans up to 10% (2 HRIPTs).

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

From September 17 to October 18, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP study performed according to OECD test guideline No. 406 with deviations. Rationale for dose-selection (max. conc.) is not reported even if the physical state of the substance is expected to be the limiting factor.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

(Adopted in May 1981)

Deviations:

yes

Remarks:

Topical induction duration was 24 hours instead of 48 hours. FCA/saline was used in the 3rd control injection instead of ethanol FCA/saline

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

(Adopted in March 1984)

Deviations:

yes

Remarks:

Topical induction duration was 24 hours instead of 48 hours. FCA/saline was used in the 3rd control injection instead of ethanol FCA/saline

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Study conducted before the adoption of the LLNA OECD Test Guideline

Species:

guinea pig

Strain:

other: Ibm: GoHI; SPF quality (Himalayan spotted)

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wölferstrasse 4. CH-4414 Füllinsdorf.
- Age at study initiation: M: 7 weeks; F: 8 weeks.
- Weight at study initiation: M: 370-394 g; F: 374-393 g.
- Housing: individually, in Makrolon type-3 cages with standard softwood bedding.
- Diet: pelleted standard Kliba 342, Batch 59/0 guinea-pig breeding / maintenance diet, ad libitum. Not contaminated.
- Water: community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid via the drinking water. Not contaminated.
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12. Music during the light period.

IN-LIFE DATES: From: 1990-09-17 To: 1990-10-28

Route:

intradermal

Vehicle:

other: ethanol

Concentration / amount:

5 %

Day(s)/duration:

D1

Adequacy of induction:

other: assumed to be highest technically applicable concentration

Route:

epicutaneous, occlusive

Vehicle:

other: ethanol

Concentration / amount:

30%

Day(s)/duration:

D8 / 24 hrs

Adequacy of induction:

other: assumed to be highest technically applicable concentration, skin pre-treated with 10% SLS

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

other: ethanol

Concentration / amount:

30, 10 and 3%

Day(s)/duration:

D22 / 24 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

10/sex/test group, 5/sex/control group

Details on study design:

RANGE FINDING TESTS:
- Intradermal injections: 2 animals; concentrations of 5, 3 and 1% in ethanol (0.1 mL/site). Assessment of dermal reactions 24 hours later. The concentration selected for the main study was 5%.
- Epidermal application: 4 animals; concentrations of 30, 10, 3 and 1% in ethanol. Assesment of dermal reactions immediately after patch removal, 24 and 48 hours later. The concentration selected for epidermal application was 30%.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: intradermal + epicutaneous.
- Test groups:
Intradermal injections: 3 pairs of intradermal injections (0.1 mL/site) made at the border of a 4 x 6 cm area in the clipped region:
1/ FCA 50:50 with physiological saline
2/ Test article, diluted to 5% with ethanol
3/ Test article diluted to 5% by emulsion in a 50:50 mixture of FCA and physiological saline.
Epidermal applications: on day 7, the test area was pre-treated with 10% Sodiul-Lauryl-Sulfate (SLS) in petrolatum oil, because no irritation concentration could be determined in the corresponding pre-test. On day 8, patches of filter paper (2 x 2 cm) were saturated with concentration of 30% test article in ethanol and applied to the clipped and shaved flanks of each animal. The patches were covered by a strip of aluminium foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. Reactions sites were assessed immediately, 24 and 48 hours after removal of the dressing.
- Control group:
Intradermal injections: 3 pairs of intradermal injections (0.1 mL/site) made at the border of a 4 x 6 cm area in the clipped region:
1/ FCA 50:50 with physiological saline
2/ Ethanol
3/ FCA 50:50 with physiological saline.
Epidermal applications: treated as described above with the omission of test article (ethanol only).
- Site: dorsal skin from the scapular region

B. CHALLENGE EXPOSURE
- No. of exposures: 1, epicutaneous
- Day(s) of challenge: 2 weeks after the epidermal induction application
- Exposure period: 24 hours
- Test groups: patches (2 x 2 cm) of filter paper were saturated with the test article and applied to the left flank using the same method as for the epidermal application.
- Control group: patches (2 x 2 cm) of filter paper were saturated with the vehicle and applied to the right flank using the same method as for the epidermal application.
- Site: left and right flanks
- Concentrations: 0, 3, 10 and 30 %
- Evaluation (hr after challenge): 24, 48 and 72 (0, 24 and 48 after patch removal)

OTHER: Erythema and oedema were assessed using the numerical grading system of Draize.

Challenge controls:

None

Positive control substance(s):

not required

Remarks:

a control group (Formaldehyde-solution) is tested twice a year for sensitivity check of the guinea-pig.

Positive control results:

The most recent test was run from April 23 to May 24, 1990. Clear positive results were observed in 7/10 Formaldehydlosung (HCHO) treated animals after the epidermal challenge application.

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

30 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

30 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

10 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

10 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

3 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

3 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

2nd reading

Hours after challenge:

72

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

other: Positive control most recent test

Group:

positive control

Dose level:

NA

No. with + reactions:

7

Total no. in group:

10

Remarks on result:

positive indication of skin sensitisation

No death occurred during the study. No systemic symptoms were observed in the animals. The body weight gain of the animals was not affected during the study.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test material did not produce evidence of skin sensitization (delayed contact hypersensitivity).

Executive summary:

A dermal sensitization study was performed according to OECD guideline No. 406 and in compliance with GLP with the test material diluted in ethanol. Male and female Himalayan spotted guinea-pigs were tested using the Guinea pig maximisation method (20 treated animal per concentration + 10 control animals).

The preliminary study determined the lowest irritant test substance concentration used at induction phases and the highest non-irritant test substance concentration used at challenge.

The test material diluted in ethanol at 5% was administered by injection for intradermal induction. On day 7, as the substance was not a skin irritant, the site was pre-treated with 10% sodium lauryl sulphate in petrolatum-oil. 24 -hour topical induction was performed with the test material diluted in ethanol at 30 % on Day 8. For the challenge, the test material was applied topically during 24 hours at 30, 10 and 3 % in ethanol.

 

The sensitivity of the guinea-pig was checked every 6 months at RCC with a Formaldehyde-solution, a known sensitizer.

 

No signs of ill health or toxicity were recorded in any animals during the observation period.

The test material did not produce evidence of skin sensitization (delayed contact hypersensitivity).

 

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

This study is considered as acceptable within a weight of evidence approach to satisfy the requirement for sensitisation endpoint.

 

Reference 2

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

From 2012-10-23 to 2012-11-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study performed according to OECD test guideline No. 429 and in compliance with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP Compliance Monitoring Programme (inspected on 2012-07-10/ signed on 2012-11-30)

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 15-23 g
- Housing: Individually housed, in suspended solid-floor polypropylene cages furnished with softwood woodflakes
- Diet: : ad libitum (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK).
- Water: mains tap water ad libitum.
The diet and water are routinely analysed and are considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

10%, 5% and 2.5%

No. of animals per dose:

5

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: maximum attainable concentration = 10%
- Irritation: No signs of systemic toxicity, local irritation or irritation indicated by ≥25% increase in mean ear thickness noted at the maximum attainable concentration).
Based on this information the dose levels selected for the main test were 10%, 5% and 2.5% w/w in acetone/olive oil 4:1.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: individual approach, using tritiated (3H)-methyl thymidine, according to the OECD 429 test guideline.
- Additional investigation: measurement of ear thickness. The ear thickness of each ear was measured using an Oditest micrometer (Dyer, PA) immediately prior to and 1-hour post application of test item on Day 1 and 1-hour post application of the test item on Days 2 & 3. Additional measurements will be taken on Days 4, 5 & 6 (Day of lymph node harvest for the LLNA) to monitor for any changes in ear thickness. Group mean ear thickness changes were calculated between Days 1 to 3 and Days 1 to 6. A mean ear thickness of ≥ 25% was considered to indicate excessive irritation and limited biological relevance to the endpoint of sensitization.
- Criteria used to consider a positive response: The decision process with regard to identification of a positive response will include a SI of ≥ 3, together with consideration of dose-response, and where appropriate, statistical significance. Any test item failing to produce a SI of ≥ 3 at all test concentrations was not regarded as a skin sensitiser.

TREATMENT PREPARATION AND ADMINISTRATION:
All formulations were used within two hours of preparation and will be assumed to be stable for this period. The concentration and homogeneity of the formulations were not determined by analysis.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Data was processed to give group mean values for disintegrations per minute and standard deviations where appropriate. Individual and group mean disintegrations per minute values were assessed for dose response relationships by analysis of homogeneity of variance followed by one way analysis of variance (ANOVA). In the event of a significant result from the ANOVA, pairwise comparisons were performed between control and treated groups. For homogenous datasets Dunnett’s Multiple Comparison test was used and for non homogenous datasets Dunnett’s T3 Multiple Comparison Method was used.

Positive control results:

A group of five animals was treated with 50 µl (25 µl per ear) of α-Hexylcinnamaldehyde, tech., 85% as a solution in acetone/olive oil 4:1 at a concentration of 25% v/v. A further control group of five animals was treated with acetone/olive oil 4:1 alone. With a SI = 6.29, α-Hexylcinnamaldehyde, tech., 85% was considered to be a sensitiser under the conditions of the test.

Key result

Parameter:

SI

Value:

0.7

Test group / Remarks:

2.5%

Key result

Parameter:

SI

Value:

1.46

Test group / Remarks:

5%

Key result

Parameter:

SI

Value:

1.09

Test group / Remarks:

10%

Cellular proliferation data / Observations:

CELLULAR PROLIFERATION DATA
0% : mean DPM / animal = 2918.93.
2.5%: mean DPM / animal = 2050.60.
5%: mean DPM / animal = 4271.47.
10%: mean DPM / animal = 3183.67

CLINICAL OBSERVATIONS: There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test. No visual local skin irritation or excessive irritation indicated by an equal to or greater than 25% increase in mean ear thickness were noted at any dose concentration evaluated.

BODY WEIGHTS: Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, ST 10 C 08 is not classified as a skin sensitiser according to the annex I of the Regulation EC No. 1272/2008 (CLP).

Executive summary:

A study was performed to assess the skin sensitisation potential of ST 10 C 08 in the CBA/Ca strain mouse following topical application to the dorsal surface of the ear. The method was conducted according to the OECD test guideline No 429 and in compliance with GLP.

Following a preliminary screening test in which no clinical signs of toxicity were noted at the maximum attainable concentration (10%), ST 10 C 08 concentration of 10% was selected as the highest dose to be investigated in the main test.

Three groups, each of five animals, were treated with 50 μl (25 μl per ear) of ST 10 C 08 as a solution in acetone/olive oil 4:1 at concentrations of 10%, 5% or 2.5% v/v for 3 consecutive days. A further group of five animals was treated with acetone/olive oil 4:1 alone. The animals were allowed to rest without dosing on days 4 and 5. On day 6, the proliferation of lymphocytes in the lymph node draining the application site was measured by incorporation of ³H-Methyl Thymidine.

The irritant potential of the test item was assessed in parallel by measurement of ear thickness on days 1 to 6.

The Stimulation Index values expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as follows:

Concentration	Mean dpm/animal	Test / Control Ratio	Result
Vehicle	2918.93	N/A	N/A
2.5%	2050.60	0.70	Negative
5%	4271.47	1.46	Negative
10%	3183.67	1.09	Negative

The historical positive control, α-Hexylcinnamaldehyde, gave a SI of 6.29, when tested at 25 % v/v. The test system was therefore considered to be valid.

There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test. No visual local skin irritation or excessive irritation indicated by an equal to or greater than 25% increase in mean ear thickness were noted at any dose concentration evaluated.

Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period.

Under the test conditions, ST 10 C 08 is not classified as a skin sensitiser in the Local Lymph Node Assay according to the annex I of the Regulation (EC) No. 1272/2008 (CLP).

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

Reference 3

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
[Further information is included as attachment to Iuclid section 13]

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across approach is based on the hypothesis that the source and target substances have similar physico-chemical, toxicological, ecotoxicological and environmental fate properties because of their structural similarity.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The target and the source substances (Table 1) are Cycloalkane ethers. The source substance is the racemic form, while the target substance is the (-)-isomer.

3. ANALOGUE APPROACH JUSTIFICATION
Based on structural similarity and comparable physicochemical and toxicological properties, the source and the target substances are expected to have similar skin sensitisation profile. This hypothesis is supported by the GPMT study result on the target substance itself (See Table 4). The reliability score of the GPMT test performed on the target substance was restricted to 2 because the rationale for dose-selection (max. conc.) was not reported even if the physical state of the substance was expected to be the limiting factor. Therefore, a read-across to the source substance was used to strengthen the conclusion on this endpoint.
The study performed on the source substance (OECD 429, GLP) is adequate and reliable for the purpose of the prediction based on read-across. The test material used represents the source substance as described in the hypothesis in terms of purity and impurities. The results of the studies are adequate for the purpose of classification and labelling.
Therefore, based on the considerations above, it can be concluded that the results of the skin sensitisation study conducted with the source substance are likely to predict the properties of the target substance and are considered as adequate to fulfil the information requirement of Annex VIII, 8.7.2.

4. DATA MATRIX
See Iuclid section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across: supporting information

Positive control results:

A group of five animals was treated with 50 µl (25 µl per ear) of α-Hexylcinnamaldehyde, tech., 85% as a solution in acetone/olive oil 4:1 at a concentration of 25% v/v. A further control group of five animals was treated with acetone/olive oil 4:1 alone. With a SI = 6.29, α-Hexylcinnamaldehyde, tech., 85% was considered to be a sensitiser under the conditions of the test.

Key result

Parameter:

SI

Value:

0.7

Test group / Remarks:

2.5%

Key result

Parameter:

SI

Value:

1.46

Test group / Remarks:

5%

Key result

Parameter:

SI

Value:

1.09

Test group / Remarks:

10%

Cellular proliferation data / Observations:

CELLULAR PROLIFERATION DATA
0% : mean DPM / animal = 2918.93.
2.5%: mean DPM / animal = 2050.60.
5%: mean DPM / animal = 4271.47.
10%: mean DPM / animal = 3183.67

CLINICAL OBSERVATIONS: There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test. No visual local skin irritation or excessive irritation indicated by an equal to or greater than 25% increase in mean ear thickness were noted at any dose concentration evaluated.

BODY WEIGHTS: Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results on the source substance ST 10 C 08, the target substance is not classified as a skin sensitiser according to the annex I of the Regulation EC No. 1272/2008 (CLP).

Executive summary:

A study was performed to assess the skin sensitisation potential of the source substance ST 10 C 08 in the CBA/Ca strain mouse following topical application to the dorsal surface of the ear. The method was conducted according to the OECD test guideline No 429 and in compliance with GLP.

Following a preliminary screening test in which no clinical signs of toxicity were noted at the maximum attainable concentration (10%), ST 10 C 08 concentration of 10% was selected as the highest dose to be investigated in the main test.

Three groups, each of five animals, were treated with 50 μl (25 μl per ear) of ST 10 C 08 as a solution in acetone/olive oil 4:1 at concentrations of 10%, 5% or 2.5% v/v for 3 consecutive days. A further group of five animals was treated with acetone/olive oil 4:1 alone. The animals were allowed to rest without dosing on days 4 and 5. On day 6, the proliferation of lymphocytes in the lymph node draining the application site was measured by incorporation of ³H-Methyl Thymidine.

The irritant potential of the test item was assessed in parallel by measurement of ear thickness on days 1 to 6.

The Stimulation Index values expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as follows:

Concentration	Mean dpm/animal	Test / Control Ratio	Result
Vehicle	2918.93	N/A	N/A
2.5%	2050.60	0.70	Negative
5%	4271.47	1.46	Negative
10%	3183.67	1.09	Negative

The historical positive control, α-Hexylcinnamaldehyde, gave a SI of 6.29, when tested at 25 % v/v. The test system was therefore considered to be valid.

There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test. No visual local skin irritation or excessive irritation indicated by an equal to or greater than 25% increase in mean ear thickness were noted at any dose concentration evaluated.

Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period.

Based on the results on the source substance ST 10 C 08, the target substance is not classified as a skin sensitiser in the Local Lymph Node Assay according to the annex I of the Regulation (EC) No. 1272/2008 (CLP).

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:

Several skin sensitisation studies were identified. Although some of the studies provided information to assess the skin sensitisation potential of the test substance, none of them was considered as sufficient as the key study on its own. Therefore, a weight-of-evidence approach was built, using the most reliable study (RCC, 1991, Rel.2) together with studies performed for the racemic form of the registered substance (Harlan, 2013, Rel.2):

- In a Guinea-pig maximisation test (GPMT) performed by RCC according to OECD guideline No. 406 and in compliance with GLP, the test material diluted in ethanol at 5% was administered by injection for intradermal induction. On day 7, as the substance was not a skin irritant, the site was pre-treated with 10% sodium lauryl sulphate in petrolatum-oil. Topical induction was performed with the test material diluted in ethanol at 30 % Day 8. For the challenge, the test material was applied topically at 30, 10 and 3 % in ethanol. The rationale for dose-selection (max. conc.) was not reported in the report even if the physical state of the substance is expected to be the limiting factor. The sensitivity of the guinea-pig was checked every 6 months at RCC with a Formaldehyde-solution, a known sensitizer. No signs of ill health or toxicity were recorded in any animals during the observation period. The test material did not produce evidence of skin sensitization (delayed contact hypersensitivity).

- In a Local Lymph Node Assay (LLNA) performed by Harlan according to the OECD test guideline No 429 and in compliance with GLP, groups of mice were treated with the source substance ST 10 C 08 (see Iuclid section 13 for read-across justification) as a solution in acetone/olive oil 4:1 at concentrations of 10% (maximum achievable concentration), 5% or 2.5% v/v. The irritant potential of ST 10 C 08 was assessed in parallel by measurement of ear thickness on days 1 to 6. The Stimulation Index values were 0.70, 1.46 and 1.09 for the concentrations of 2.5%, 5% and 10%, respectively. The historical positive control, α-Hexylcinnamaldehyde, gave a SI of 6.29, when tested at 25 % v/v. The test system was therefore considered to be valid. There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test. No visual local skin irritation or excessive irritation indicated by an equal to or greater than 25% increase in mean ear thickness were noted at any dose concentration evaluated. Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period. Under the test conditions, the source substance ST 10 C 08 is not classified as a skin sensitiser.

Although poorly reliable, a Buehler test (Huntington, 1971, Rel.3) and an Open Epidermal test (Henkel, 1993, Rel.3) support the conclusion.

Two human repeated insult patch tests in human have also been performed with the substance at up to 10 % (TKL, 2003 and Biosearch 1979). Both studies were concluded to be negative for sensitisation.

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 I of the Regulation (EC) No. 1272/2008 (CLP).

No data was available regarding respiratory sensitisation.