Registration Dossier

Administrative data

Description of key information

Weight of evidence: The skin sensitization potential of the main constituents are available from experimental data. Based on these experimental data and since content of those classified as skin sensitizers 1B is higher than 1%, it is determined that the substance is classified as skin sensitizer 1B, H317 according to the CLP Regulation (EC) no. 1272/2008.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
Lack of data on test animals and housing conditions.
GLP compliance:
not specified
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
not specified
Sex:
not specified
Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
0 (vehicle), 1, 5 10, 15 and 25% w/v
No. of animals per dose:
three females
Details on study design:
MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: murine local lymph node assay.
- Criteria used to consider a positive response:
Test materials that at one or more concentrations gave rise to an stimulation index (SI) greater than 3 were considered to be sensitizers in the LLNA.
The sensitizing potency of the test compound was classified according to the following: <0.1, extreme; ≥0.1-<1, strong; ≥1- <10, moderate; and ≥10-<100, weak ((ECETOC (2003), Kimber et al. (2003)).
Positive control substance(s):
other: The results of this study were compared with the prohaptenic cinnamic alcohol and its reactive metabolite cinnamic aldehyde.
Statistics:
Results were expressed as mean dpm/lymph node for each experimental group and as stimulation index (SI), i.e., test group/control group ratio.
EC3 values were calculated by linear interpolation (Basketter et al. (1999)) and used to determine the sensitizing potency of the test substance.

Positive control results:
Prohaptenic cinnamic alcohol and its reactive metabolite cinnamic aldehyde gave EC3 values of 1.5 (20.1%) and 0.10 M (1.3%), respectively (Elahi et al. (2004)).
Key result
Parameter:
SI
Value:
1.1
Test group / Remarks:
1 % w/v
Key result
Parameter:
SI
Value:
1.5
Test group / Remarks:
5 % w/v
Key result
Parameter:
SI
Value:
3.4
Test group / Remarks:
10 % w/v
Key result
Parameter:
SI
Value:
8.9
Test group / Remarks:
15 % w/v
Key result
Parameter:
SI
Value:
23
Test group / Remarks:
25% w/v
Key result
Parameter:
EC3
Value:
8.9
Remarks on result:
other: EC3= estimated concentration to induce a Stimulation index of 3

Table 1. LLNA results for alpha terpinene

Alpha terpinene and test concentrations

(% w/v)

[3H]thymidine

incorporation

(dpm/lymph node)

SI

 

EC3 value of  

Alpha terpinene and test concentrations

(% w/v)

sensitizing potency

% w/v

M

0

755

 

8.9

0.65

moderate

1

848

1.1

5

1153

1.5

10

2595

3.4

15

6740

8.9

25

17176

23

Interpretation of results:
other: Category 1B (CLP Regulation EC no. 1272/2008)
Conclusions:
Based on the results of this study, alpha terpinene was shown to be sensitizer of moderate potency with the EC3 value of 0.65 M (8.9 % w/v). The SI was greater than 3 at the concentrations of 10, 15 and 25 %.
Executive summary:

The murine LLNA assay was used in this study to assess the sensitizing potency of alpha terpinene which was tested at concentrations of 0 (acetone/olive oil, 4:1 v/v), 1, 5, 10, 15 and 25 (% w/v) using mice in groups of three. The results were expressed as mean dpm/lymph node for each experimental group and as stimulation index (SI), i.e., test group/control group ratio. Test material that at one or more concentrations gave rise to an SI greater than 3 were considered to be sensitizers in the LLNA. The results of this study were compared with the prohaptenic cinnamic alcohol and its reactive metabolite cinnamic aldehyde, which have EC3 values of 1.5 (20.1%) and 0.10 M (1.3%), respectively. The results demonstrated that the SI was greater than 3 at the concentrations of 10, 15 and 25 % of alpha terpinene. Based on these results, alpha terpinene was shown to be a sensitizer of moderate potency with EC3 value of 0.65 M (8.9 % w/v).

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
maximization test
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline available
Principles of method if other than guideline:
A maximization test (Kligman, 1966; Kligman & Epstein, 1975) was carried out on 25 volunteers.
No more data was provided on the method.
GLP compliance:
no
Type of study:
other: maximization test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Reading:
1st reading
Group:
test group
Dose level:
4 % in petrolatum
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

The material was tested at a concentration of 4 % in petrolatum and produced no sensitization reactions.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The material was tested on humans at a concentration of 4 % in petrolatum and produced no sensitization reactions.
Executive summary:

A maximization test was carried out on 25 volunteers. The material was tested at a concentration of 4 % in petrolatum and produced no sensitization reactions.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
maximization test
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline available
Principles of method if other than guideline:
A maximization test (Kligman, 1966; Kligman & Epstein, 1975) was carried out on 25 volunteers.
No more data was provided on the method.
GLP compliance:
no
Type of study:
other: maximization test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Reading:
1st reading
Group:
test group
Dose level:
5 % in petrolatum
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

The material was tested at a concentration of 5 % in petrolatum and produced no sensitization reactions.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The material was tested on humans at a concentration of 5 % in petrolatum and produced no sensitization reactions.
Executive summary:

A maximization test was carried out on 25 volunteers. The material was tested at a concentration of 5 % in petrolatum and produced no sensitization reactions.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
maximization test
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline available
Principles of method if other than guideline:
-Principle of test: A maximization test (Kligman, 1966; Kligman & Epstein, 1975) was carried out on 24 volunteers.
GLP compliance:
no
Type of study:
other: maximization test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Reading:
1st reading
Group:
test group
Dose level:
20% in petrolatum
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

The material was tested at a concentration of 20 % in petrolatum and produced no sensitization reactions.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The material was tested on humans at a concentration of 20 % in petrolatum and produced no sensitization reactions.
Executive summary:

A maximization test was carried out on 24 volunteers. The material was tested at a concentration of 20 % in petrolatum and produced no sensitization reactions.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
maximization test
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline available
Principles of method if other than guideline:
A maximization test (Kligman, 1966; Kligman & Epstein, 1975) was carried out on 25 volunteers.
No more data was provided on the method.
GLP compliance:
no
Type of study:
other: maximization test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Key result
Reading:
1st reading
Group:
test group
Dose level:
10 % in petrolatum
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

The material was tested at a concentration of 10 % in petrolatum and produced no sensitization reactions.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The material was tested on humans at a concentration of 10% in petrolatum and produced no sensitization reactions.
Executive summary:

A maximization test was carried out on 25 volunteers. The material was tested at a concentration of 10% in petrolatum and produced no sensitization reactions.

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
not specified
Remarks:
No information is provided on the report.
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: Scanbur or NOVA SCB/Charles River (Sollentuna, Sweden or Germany, respectively)
- Age at study initiation: around 8 weeks
Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
0 (vehicle), 0.1, 1, 5 10 and 25% w/v (3 weeks air exposure assay)
0 (vehicle), 1, 5 10, 15 and 30% w/v (7 weeks air exposure assay)
No. of animals per dose:
three females
Details on study design:
MAIN STUDY
Groups of female CBA/Ca mice (n = 3) received 25 µL of the test compound dissolved in vehicle (acetone/olive oil 4:1, v/v), on the dorsum of the ears daily for 3 consecutive days. Control animals were treated with a vehicle alone.

All mice were injected intravenously 5 days after the first treatment, with 250 µL of phosphate-buffered saline (PBS) containing 20 µCi of [3H]thymidine. After 5 h, the draining auricular lymph nodes were excised and pooled for each group, and single cell suspensions of lymph node cells were prepared. The relative thymidine incorporation was measured by ß-scintillation counting. Results were expressed as mean dpm/lymph node for each experimental group and as a stimulation index (SI), i.e., test group/control group ratio.

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: murine local lymph node assay.
- Criteria used to consider a positive response:
Test materials that at one or more concentrations gave rise to an stimulation index (SI) greater than 3 were considered to be sensitizers in the LLNA.
The sensitizing potency of the test compound was classified according to the following: <0.1, extreme; ≥0.1-<1, strong; ≥1- <10, moderate; and ≥10-<100, weak ((ECETOC (2003), Kimber et al. (2003)).

Positive control substance(s):
other: The results of this study were compared with the prohaptenic cinnamic alcohol and its reactive metabolite cinnamic aldehyde.
Statistics:
Results were expressed as mean dpm/lymph node for each experimental group and as stimulation index (SI), i.e., test group/control group ratio.
EC3 values were calculated by linear interpolation (Basketter et al. (1999)) and used to determine the sensitizing potency of the test substance.

Parameter:
SI
Value:
0.8
Test group / Remarks:
0.1 (%w/v)
Remarks on result:
other: 3 week air exposure assay
Parameter:
SI
Value:
3.2
Test group / Remarks:
1 (% w/v)
Remarks on result:
other: 3 week air exposure assay
Parameter:
SI
Value:
13
Test group / Remarks:
5 (% w/v)
Remarks on result:
other: 3 week air exposure assay
Parameter:
EC3
Value:
0.9
Remarks on result:
other: 3 week air exposure assay
Parameter:
SI
Value:
3
Test group / Remarks:
1 (% w/v)
Remarks on result:
other: 7 week air exposure assay
Parameter:
SI
Value:
10
Test group / Remarks:
5 (% w/v)
Remarks on result:
other: 7 week air exposure assay
Parameter:
SI
Value:
13
Test group / Remarks:
10 (% w/v)
Remarks on result:
other: 7 week air exposure assay
Parameter:
EC3
Value:
1
Remarks on result:
other: 7 week air exposure assay

Table 1. LLNA Responses for 3-week air-exposed alpha terpinene

air-exposed alpha terpinene(% w/v)

[3H]thymidine

incorporation

(dpm/lymph node)

SI

 

EC3 value ofair-exposed alpha terpinene(% w/v)

sensitizing potency

% w/v

M

Control

1075

 

0.9

 

 

strong

0.1

857

0.8

1

3380

3.2

5

14168

13

10

18399

17

25

13365

12

Table 2. LLNA Responses for 7-week air-exposed alpha terpinene

air-exposed alpha terpinene(% w/v)

[3H]thymidine

incorporation

(dpm/lymph node)

SI

 

EC3 value ofair-exposed alpha terpinene(% w/v)

sensitizing potency

% w/v

M

Control

1290

 

1

 

 

strong

1

3900

3.0

5

13074

10

10

16781

13

15

10593

8.2

25

12074

9.4

Interpretation of results:
study cannot be used for classification
Conclusions:
The results demonstrated that alpha terpinene is able to auto-oxidise rapidly on exposure to air and to acquire strong skin sensitising potential, with an EC3 of approximately 1%.


Executive summary:

The murine LLNA assay was used in this study to assess the sensitizing potency of air exposed alpha terpinene and its oxidation products. A sample of pure alpha terpinene was air-exposed (3 week and 7 weeks) at room temperature and under a daylight lamp lit for 12 h a day. The oxidation mixture was stirred four times daily for 1 h. Samples were taken on a regular basis and analysed using LC/UV or LC/MS/MS for quantification of alpha terpinene and its major oxidation products. It was shown, that the concentration of alpha terpinene decreased rapidly when exposed to air at room temperature. The concentration of alpha terpinene decreased to 53% after 10 days and 21% after 24 days, and after 66 days, alpha terpinene was not detected in the oxidation mixture (method limit of detection 0.2%). Groups of female CBA/Ca mice (n = 3) received 25 µL of the test compound dissolved in vehicle (acetone/olive oil 4:1, v/v), on the dorsum of the ears daily for 3 consecutive days. Control animals were treated with a vehicle alone. All mice were injected intravenously 5 days after the first treatment, with 250 µL of phosphate-buffered saline (PBS) containing 20 µCi of [3H]thymidine. After 5 h, the draining auricular lymph nodes were excised and pooled for each group, and single cell suspensions of lymph node cells were prepared. The relative thymidine incorporation was measured by ß-scintillation counting. Results were expressed as mean dpm/lymph node for each experimental group and as stimulation index (SI), i.e., test group/control group ratio. The results of the study showed that alpha terpinene is able to auto-oxidise rapidly on exposure to air, and to acquire strong skin sensitising potential, with an EC3 of approximately 1%. The following EC3 values were afforded for air exposed alpha terpinene: three weeks oxidized alpha terpinene: 0.9% w/v (content of pure alpha terpinene: ca. 20%); seven weeks oxidized alpha terpinene: 1.0% w/v (content of pure alpha terpinene: ca. 2%).

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2001
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
no data on physicochemical properties of test substance; no data on individual weights, housing conditions of test animals; no information on time course of onset and signs of toxicity
GLP compliance:
not specified
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Seralab, Oxon, UK
- Age at study initiation: 8-12 weeks
- Housing: four/cage on flushing metal racks
- Diet: SDS PCD pelleted diet; Special Diets Services, Witham, UK; ad libitum
- Water: ad libitum

Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
0 (vehicle), 25, 50 and 100% v/v
No. of animals per dose:
Four females
Details on study design:
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local lymph node assay (Kimber and Basketter, 1992)
- Criteria used to consider a positive response: Stimulation index (SI) ≥ 3 (Kimber and Basketter, 1992).

TREATMENT PREPARATION AND ADMINISTRATION:
Groups of mice received 25 µL of various concentrations of limonene in vehicle, or vehicle alone, on the dorsum of both ears daily for 3 consecutive days. Five days following the initiation of treatment, all mice were injected intravenously with 250 µL of 20 µCi 3H-TdR in PBS. Five hours later draining auricular lymph nodes were excised and a single cell suspension prepared. The incorporation of 3H-TdR was measured by β-scintillation counter (dpm/node and stimulation index [SI]) (Kimber and Basketter, 1992).

The estimated concentration of chemical required to induce a stimulation index of 3 relative to concurrent vehicle-treated controls (EC3 value) was derived by linear interpolation by method as explained by Basketter et al. (1999).

The EC3 value was calculated by interpolating between two points on the SI axis, one immediately above, and the other immediately below, the SI value of three. The vehicle-treated control value (SI=1) cannot be used for the latter. Where the data points falling immediately above and below the SI value of three have the co-ordinates (a,b) and (c,d), respectively, then the EC3 value may be calculated using the following equation (Basketter et al., 1999): EC3=c + [3−d) / (b−d)] × (a−c).
Statistics:
None
Key result
Parameter:
SI
Value:
1
Test group / Remarks:
0 % v/v
Key result
Parameter:
SI
Value:
1.84
Test group / Remarks:
25 % v/v
Key result
Parameter:
SI
Value:
2.44
Test group / Remarks:
50 % v/v
Key result
Parameter:
SI
Value:
3.95
Test group / Remarks:
100 % v/v
Key result
Parameter:
EC3
Value:
68.5
Remarks on result:
other: EC3= estimated concentration to induce a Stimulation index of 3

Table 1: Local lymph node assay responses to limonene

Exposure Concentration (% v/v)

dpm/node

Stimulation index (SI)

EC3 (%)

0

476

1

68.5

25

877

1.84

50

1164

2.44

100

1882

3.95

Interpretation of results:
other: Category 1B (CLP Regulation EC no. 1272/2008)
Conclusions:
The calculated EC3-value for d-limonene was found to be 68.5%. Under these test conditions, d-limonene was found to be sensitising and should be clasified as skin sensitiser 1B according to the criteria of the CLP Regulation (EC) N° (1272-2008).
Executive summary:

In a skin sensitization study conducted following a method similar to OECD guidelines 429, four groups of CBA/Ca strain female mice (4/concentration) were exposed topically on the dorsum of both ears to 25 µL of vehicle (acetone/olive oil, 4:1 v/v), 25, 50 and 100% v/v of d-limonene daily for 3 consecutive days. Five days following initiation of exposure all mice were injected 250 µL of phosphate buffered saline (PBS) containing 20 µCi of [3H] methyl thymidine (3H-TdR). The animals were then sacrificed after 5 hours and suspension of lymph node cells was prepared from auricular lymph nodes. The incorporation of 3H-TdR was measured by β-scintillation counting as disintegrations per minute (dpm) per node for each experimental group. A Stimulation index of 3 or greater was considered to be indicative of a potential to cause contact sensitization. D-limonene at exposure concentration of 0 (vehicle), 25, 50 and 100% v/v resulted in 476, 877, 1164 and 1882 dpm/node and stimulation index of 1, 1.84, 2.44 and 3.95, respectively. The calculated EC3-value for d-limonene was found to be 68.5%. Under these test conditions, d-limonene was found to be sensitising and should be clasified as skin sensitiser 1B according to the CLP Regulation (EC) N° (1272-2008).

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils.
No more data provided on the method.
GLP compliance:
no
Type of study:
patch test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
no data
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
No data
No. of animals per dose:
No data
Positive control substance(s):
not specified
Key result
Reading:
1st reading
Group:
test group
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

alpha pinene was found not to be a sensitizer for human skin.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
Alpha pinene was found not to be a sensitizer for human skin.
Executive summary:

A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils. Alpha pinene was found not to be a sensitizer for human skin.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils.
No more data provided on the method.
GLP compliance:
no
Type of study:
patch test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
no data
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
No data
No. of animals per dose:
No data
Positive control substance(s):
not specified
Reading:
1st reading
Group:
test group
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

Camphene was found not to be a sensitizer for human skin.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
Camphene was found not to be a sensitizer for human skin.
Executive summary:

A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils. Camphene was found not to be a sensitizer for human skin.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
-Principle of test: A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils. No more data provided on the method.
GLP compliance:
no
Type of study:
patch test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
no data
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
No data
No. of animals per dose:
No data
Positive control substance(s):
not specified
Reading:
1st reading
Group:
test group
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: no data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

Terpinolene was found not to be a sensitizer for human skin.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
Terpinolene was found not to be a sensitizer for human skin.
Executive summary:

A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils. Terpinolene was found not to be a sensitizer for human skin.

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
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
In accordance with column 2 of REACH Annex VII, the study does not need to be conducted since in vivo studies on sensitisation are available.
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
- Principle of test: Rat popliteal lymph node assay (PLNA). Although the predictive value of PLNA was initially focused on auto-immune-like reactions, more recently it has been regarded as a screening test useful for detecting a broader class of sensitizing or immuno-stimulating chemicals (Pieters and Albers, 1999; Ravel and Descotes, 2005). In this study, the rat PLNA was used to evaluate the immuno-sensitizing potential of 10 monoterpenes found in the essential oils of a variety of aromatic, edible and medicinal plants.

GLP compliance:
not specified
Type of study:
other: Popliteal lymph node assay (PLNA)
Justification for non-LLNA method:
The PLNA seems to be a reliable test for screening chemicals causing sensitization via routes of exposure other than the skin (Goebel et al., 1996; Tuschl et al., 2002).
Species:
other: rat
Strain:
other: Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Oswaldo Cruz Foundation (FIOCRUZ) breeding stock
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 7–8 week-old
- Weight at study initiation: 145 ± 23 g
- Housing: All rats were individually housed in standard plastic cages with stainless steel cover lids and wood shavings as bedding.
- Diet (e.g. ad libitum): ad libitum (Nuvitall, Nuvilab, Curitiba, PR, Brazil)
- Water (e.g. ad libitum): ad libitum (tap water)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 1ºC,
- Humidity (%): 70%
- Photoperiod (hrs dark / hrs light): 12-h dark/light cycle
Route:
other: subcutaneous
Vehicle:
DMSO
Concentration / amount:
5 mg/paw of test substance (50 µL of test substance in vehicle)
Day(s)/duration:
7 days
Adequacy of induction:
other: It is of note that, as a rule, 5 mg/paw has been the highest dose of a test compound assayed in the rat PLNA (Vial et al., 1997; Descotes et al., 1997).
No. of animals per dose:
10
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: one per animal
- Exposure period: Seven days after footpad injection, rats were killed by CO2 inhalation
- Test groups: one treated group (10 animals)
- Control group: barbital (negative control), saline (vehicle control for negative) and DMSO (vehicle control for treated group)
- Site: right footpad (treated group). The contralateral (left) hind footpad was used as the control and thus it was injected with 50 µL of vehicle alone. When the vehicles were tested, the right hind footpad was injected with 50 µL of DMSO or saline while the left hind footpad remained untreated.

OTHER:
Seven days after footpad injection, rats were killed by CO2 inhalation. The popliteal lymph nodes (PLN) were carefully removed, placed in phosphate buffer saline (PBS), freed from adherent fatty tissue and weighed. PLNs were then transferred to a glass tube where cells were extracted by agitation and gentle grinding using a micropestle and suspended in a known volume of PBS on ice. PLN cell number (cellularity) was counted using a Neubauer chamber. Results were expressed as weight
(WI) and cellularity (CI) indices. WI and CI were calculated by dividing the value (weight or cellularity) obtained for the treated (right) side PLN by that obtained for the control (left) side PLN.

Positive control substance(s):
yes
Remarks:
(chlorpromazine)
Positive control results:
see table on "any other information on results incl. tables"
Key result
Reading:
1st reading
Group:
test group
Dose level:
5 mg
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Group:
negative control
Dose level:
5 mg
No. with + reactions:
0
Total no. in group:
8
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Group:
positive control
Dose level:
5 mg
Remarks on result:
positive indication of skin sensitisation

Table 1: Primary PLNA responses

 Criteria

Negative controls

Vehicle

Chlorpromazine

 

 Terpineol 

 Barbital 

DMSO

 Saline

0.5 mg/paw

2.5 mg/paw

5.0 mg/paw

 N 

 10 

 8 

 47 

 50 

 4 

 6 

 11 

 WI 

 1.32 ± 0.71 

 1.06 ± 0.36 

 1.48 ± 0.65 

 1.12 ± 0.90 

 1.30 ± 0.35 

 2.06 ± 0.81* 

 3.22 ± 1.13* 

 CI 

 2.00 ± 3.32 

 1.34 ± 0.92 

 2.95 ± 3.68 

 2.04 ± 2.03 

 1.26 ± 0.81 

 8.49 ± 8.91* 

 8.28 ± 8.19* 

 IPR, no. (%) 

 2 (20) 

 0 (0) 

 5 (10.6) 

 3 (6) 

 0.0 

 50.0 

 63.6* 

* indicates that the value differs from that of the lowest dose group (0.5 mg/paw)

- Injected doses were 5 mg/paw (terpineol and barbital) or 50 µL /paw (vehicles).

- Values are means ± SD;

- Weight (WI) and Cellularity (CI) indices: values for the draining popliteal lymph node of the treated (right paw) side divided by that of the control (left paw) side.

- IPR, number (%) of rats within the group with WI ≥ 2 and CI ≥ 5.

- Terpineol was classified as negative because group mean values for WI and CI were lower than 2 and 5, respectively.

Interpretation of results:
other: No category (CLP Regulation EC no. 1272/2008)
Conclusions:
Under these test conditions, alpha-terpineol was not considered as a sensitiser.
Executive summary:

In a popliteal lymph node assay (PLNA), a group of 10 female Wistar rats were injected subcutaneously with alpha terpineol at 5 mg/paw into the right hind footpad while the contralateral footpad was injected with the vehicle (DMSO) alone. Chlorpromazine (CPZ) and barbital were used as positive and negative controls, respectively. Weight (WI) and cellularity (CI) indices for draining PLNs were determined 7 days after treatment. Weight (WI) and cellularity (CI) indices for alpha terpineol was determined to be 1.32 and 2.00, respectively. Alpha terpineol was classified as negative because group mean values for WI and CI were lower than 2 and 5, respectively. Under these test conditions, alpha terpineol was not considered as a sensitiser.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
modified FCA-method (Freund’s complete adjuvant) (Hausen and Angel, 1992)
Deviations:
yes
Remarks:
(no details of housing and evironmental conditions of the animals)
GLP compliance:
not specified
Type of study:
Freund's complete adjuvant test
Justification for non-LLNA method:
Test was performed before OECD LLNA guideline approval and based on reliable method according to OECD 406 guideline.
Species:
guinea pig
Strain:
other: Pirbright-white stem
Sex:
female
Route:
epicutaneous, open
Vehicle:
physiological saline
Concentration / amount:
30 mg of oxidized tea tree oil (TTO) dissolved in 4 mL of FCA and emulsified with 4 mL of physiological saline.
Day(s)/duration:
11 days
Adequacy of induction:
not specified
No.:
#1
Route:
epicutaneous, open
Vehicle:
other: acetone
Concentration / amount:
0.05 mL of 10% alpha terpineol-solution in acetone
Adequacy of challenge:
not specified
No. of animals per dose:
10
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 1
- Exposure period: 11 days
- Test groups: one treated group (10 animals)
- Concentrations: 30 mg of oxidized tea tree oil (TTO) dissolved in 4 mL of FCA and emulsified with 4 mL of physiological saline.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Test groups: one treated group (10 animals)
- Site: clipped and shaved right flank of the animals.
- Concentrations: 10% alpha terpineol-solution in acetone
- Evaluation (hr after challenge): 24, 48 and 72 hours (according to the rules of the International Contact Dermatitis Research Group (ICDRG)).


Positive control substance(s):
no
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.05 mL of 10% alpha terpineol-solution in acetone
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.05 mL of 10% alpha terpineol-solution in acetone
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Group:
negative control
Remarks on result:
other: no data
Key result
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

Table 1: Results of the Sensitization and Challenging Experiments

Nº of guinea pigs

Sensitized with

Challenged with

Concentration

Mean response

24 h

48 h

10

TTO, oxidized

Alpha terpineol

10%

0

0

Interpretation of results:
other: No category (CLP Regulation EC no. 1272/2008)
Conclusions:
Under these test conditions, alpha-terpineol was found to be a non skin sensitiser.

Executive summary:

Experimental sensitization was performed in albino guinea pigs by a modified FCA-method (Freund’s complete adjuvant). For induction, 30 mg of oxidized tea tree oil (TTO) dissolved in 4 mL of FCA and emulsified with 4 mL of physiological saline were used on 10 female guinea pigs. Eleven days after induction, open epicutaneous challenge was performed by application of 0.05 mL of 10% alpha terpineol-solution in acetone on the clipped and shaved right flank of the animals. The reactions were read at 24 hours, 48 hours, and 72 hours. As the maximum reaction was observed in most of the animals already in the 48 hour reading, the 72 hour readings were not noted. The mean responses obtained after 24 h and 48 h were 0 and thus it is concluded that alpha terpineol is not a skin sensitizer.

In a popliteal lymph node assay (PLNA), a group of 10 female Wistar rats were injected subcutaneously with alpha terpineol at 5 mg/paw into the right hind footpad while the contralateral footpad was injected with the vehicle (DMSO) alone. Chlorpromazine (CPZ) and barbital were used as positive and negative controls, respectively. Weight (WI) and cellularity (CI) indices for draining PLNs were determined 7 days after treatment. Weight (WI) and cellularity (CI) indices for alpha terpineol was determined to be 1.32 and 2.00, respectively. Alpha terpineol was classified as negative because group mean values for WI and CI were lower than 2 and 5, respectively. Under these test conditions, alpha terpineol was not considered as a sensitiser.

Endpoint:
skin sensitisation, other
Remarks:
maximization test
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline available
Principles of method if other than guideline:
-Principle of test: A maximization test (Kligman, 1966; Kligman & Epstein, 1975) was carried out on 25 volunteers.
GLP compliance:
no
Type of study:
other: maximization test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Key result
Remarks on result:
no indication of skin sensitisation

The material was tested at a concentration of 5 % in petrolatum and produced no sensitization reactions.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The material was tested on humans at a concentration of 5 % in petrolatum and produced no sensitization reactions.
Executive summary:

A maximization test was carried out on 25 volunteers. The material was tested at a concentration of 5 % in petrolatum and produced no sensitization reactions.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
-Principle of test: A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils. No more data provided on the method.
GLP compliance:
no
Type of study:
patch test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
no data
Route:
other: No data
Vehicle:
other: No data
Concentration / amount:
No data
No. of animals per dose:
No data
Positive control substance(s):
not specified
Key result
Reading:
1st reading
Group:
test group
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

gamma terpinene was found not to be a sensitizer for human skin.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
Gamma terpinene was found not to be a sensitizer for human skin.
Executive summary:

A Patch-Test was performed on humans for stuying the sensitizing properties of 17 terpenes and related compounds present in essential oils. Gamma terpinene was found not to be a sensitizer for human skin.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
maximization test
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Qualifier:
no guideline available
Principles of method if other than guideline:
-Principle of test: A maximization test (Kligman, 1966; Kligman & Epstein, 1975) was carried out on 25 volunteers.
GLP compliance:
no
Type of study:
other: maximization test
Justification for non-LLNA method:
Data suitable for a weight of evidence approach.
Species:
other: human
Strain:
other: Not applicable
Sex:
not specified
Reading:
1st reading
Group:
test group
Dose level:
3% in petrolatum
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
negative control
Remarks on result:
other: No data
Reading:
1st reading
Group:
positive control
Remarks on result:
other: No data

The material was tested at a concentration of 3% in petrolatum and produced no sensitization reactions.

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The material was tested on humans at a concentration of 3 % in petrolatum and produced no sensitization reactions.
Executive summary:

A maximization test was carried out on 25 volunteers. The material was tested at a concentration of 3 % in petrolatum and produced no sensitization reactions.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
- Principle of test: modified Draize procedure (Draize J H, 1959)
GLP compliance:
no
Type of study:
other: modified Draize test
Justification for non-LLNA method:
An appropriate modified Draize test in guinea pigs is available which would not justify conducting an additional LLNA due to animal welfare.
Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
- Weight at study initiation: about 350 g (main test); about 450 g (preliminary irritation test)
- Housing: During testing animals were housed in wire mesh cages in pairs of the same sex.
- Diet (e.g. ad libitum): pelleted guinea pig diet, cabbage, hay ad libitum.
- Water (e.g. ad libitum): ad libitum

Route:
intradermal
Vehicle:
not specified
Concentration / amount:
0.1 mL (test substance in suitable solvent) at 2.5 x 0.25%.
Day(s)/duration:
14 days
Adequacy of induction:
other: 2.5 x concentration giving slight but perceptible irritation with no oedema when examined in a preliminary test for size (two largest diameters), erythema and oedema for 24 h (see "Details on study design")
No.:
#1
Route:
intradermal
Vehicle:
not specified
Concentration / amount:
0.1 mL (test substance in suitable solvent) at 0.25%.
Day(s)/duration:
7 days
Adequacy of challenge:
other: Concentration giving slight but perceptible irritation with no oedema when examined in a preliminary test for size (two largest diameters), erythema and oedema for 24 h (see "Details on study design")
No.:
#2
Route:
epicutaneous, open
Vehicle:
not specified
Concentration / amount:
0.1 mL (test substance in suitable solvent) at 50%.
Day(s)/duration:
7 days
Adequacy of challenge:
other: The highest concentration which caused no irritation when examined in a preliminary test for erythema for 24 h (see "Details on study design")
No. of animals per dose:
10
Details on study design:
RANGE FINDING TESTS: preliminary irritation test
Intradermal injection: Four animals of the same sex and weighing about 450 g were each injected intradermally on the shaved flanks with 0.1 mL of a range of concentrations of test material in a suitable solvent.
The reactions were examined for size (two largest diameters), erythema and oedema 24 h later and the concentration giving slight but perceptible irritation with no oedema was selected as the injection challenge concentration (ICC).

Topical application: 0.1 mL of a range of concentrations of test substance in a suitable solvent were applied in small circular areas to the shaved flanks of 4 guinea pigs of the same sex and weighing about 450 g.
The reactions were examined for erythema 24 h later and the highest concentration which caused no irritation was selected as the application challenge concentration (ACC).

MAIN STUDY
A. INDUCTION EXPOSURE
For each animal 0.1 mlL of test substance at 2.5 times the ICC were injected intradermally at 4 sites which overlie the 2 auxillary and 2 inguinal lymph nodes.
Duration: 14 days until challenge exposure.

B. CHALLENGE EXPOSURE
Each animal was challenged intradermally in one flank and topically in the other with 0.1 mL of test substance at the respective ICC and ACC: the topical application was made by spreading 0.1 mL of the test substance onto the shaved flank in a
small circular area which was not covered.

24 hours later the reactions were scored and apparent sensitization reactions confirmed 7 days later by a second challenge with controls included.

In the absence of sensitization reactions at first challenge the induction and challenge procedures were repeated, but this time a comfirmatory challenge with controls was included irrespective of any apparent sensitization reactions at the previous
challenge.

Challenge controls:
At each challenge with controls, 4 previously untreated animals of the same sex and similar weight to the test animals were treated intradermally and topically on opposite flanks with 0.1 mL of test substance at the ICC and ACC respectively.
Positive control substance(s):
no
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
24
Group:
test group
Dose level:
0.25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
24
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
0.25%
No. with + reactions:
0
Total no. in group:
4
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
4
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
4
Remarks on result:
no indication of skin sensitisation
Key result
Group:
positive control
Remarks on result:
not measured/tested
Interpretation of results:
other: No category (CLP Regulation EC no. 1272/2008)
Conclusions:
Under these test conditions, cineole was not considered as a sensitiser.
Executive summary:

The sensitization potential of cineole was tested using a modified Draize procedure with Hartley strain albino guinea pigs. Based on a preliminary irritation test, ten guinea pigs were fisrt inducted by intradermal injection of 0.1 mL of test substance diluted in suitable solvent at concentration of 2.5 x 0.25% (concentration giving slight but perceptible irritation with no oedema when examined after intradermal injection for size (two largest diameters), erythema and oedema for 24 h). After 14 days, the animals were challenged intradermally in one flank and topically in the other with 0.1 mL of test substance diluted in suitable solvent at the respective concentrations of 0.25% (same as in the induction) and 50% ( highest concentration which caused no irritation when examined after topical application for erythema for 24 h). 7 days later, as no sensitization reactions were observed, the induction and challenge procedures were repeated. Once again, no reactions were reported in these sencond readings and a rechallenge with controls were finally performed with 4 previously untreated animals which confirmed that cineole is not a skin sensitizer.

In a popliteal lymph node assay (PLNA), a group of 10 female Wistar rats were injected subcutaneously with alpha terpineol at 5 mg/paw into the right hind footpad while the contralateral footpad was injected with the vehicle (DMSO) alone. Chlorpromazine (CPZ) and barbital were used as positive and negative controls, respectively. Weight (WI) and cellularity (CI) indices for draining PLNs were determined 7 days after treatment. Weight (WI) and cellularity (CI) indices for alpha terpineol was determined to be 1.32 and 2.00, respectively. Alpha terpineol was classified as negative because group mean values for WI and CI were lower than 2 and 5, respectively. Under these test conditions, alpha terpineol was not considered as a sensitiser.

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

Weight of evidence: The skin sensitization potential of the main constituents are available from experimental data:

Alpha terpineol: Popliteal lymph node assay (Friedrich K, 2007). Alpha terpineol was not considered as a sensitiser.

Alpha terpineol: Modified FCA-method (Freund’s complete adjuvant) (Hausen B M, 1999). The test material was not found as a skin sensitizer.

Terpinolene: Patch test on humans (Woeber K, 1969). Terpinolene was found not to be a sensitizer for human skin.

Terpinolene: Maximization test on 24 volunteers (Opdyke DLJ, 1976). The material was tested on humans at a concentration of 20 % in petrolatum and produced no sensitization reactions.

Cineole: Modified Draize test (Sharp D W, 1978). Cineole was found not to be a skin sensitizer in guinea pigs.

D-Limonene: Test method similar to OECD Guideline 429 (Warbrick EV, 2001). The calculated EC3-value was found to be 68.5%, and thus, classified as skin sensitizer 1B according to the CLP Regulation (EC) no. 1272/2008.

Alpha terpinene: Maximization test on 25 volunteers (Opdyke DLJ, 1976). The material was tested on humans at a concentration of 5 % in petrolatum and produced no sensitization reactions.

Alpha terpinene: Test method similar to OECD Guideline 429 (Rudbäck J, 2012). The results demonstrated that alpha terpinene is able to auto-oxidise rapidly on exposure to air and to acquire strong skin sensitising potential, with an EC3 of approximately 1%.

Alpha terpinene: Test method similar to OECD Guideline 429 (Bergstrom MA, 2006). Alpha terpinene was shown to be sensitizer of moderate potency with the EC3 value of 0.65 M (8.9 % w/v). The SI was greater than 3 at the concentrations of 10, 15 and 25 %. According to the CLP Regulation (EC) no. 1272/2008, the substance is classified as skin sensitizer 1B.

Gamma terpinene: Patch test on humans (Woeber K, 1969). Gamma terpinene was found not to be a sensitizer for human skin.

Gamma terpinene: Maximization test on 25 volunteers (Opdyke DLJ, 1976). The material was tested on humans at a concentration of 5 % in petrolatum and produced no sensitization reactions.

Alpha pinene: Patch test on humans (Woeber K, 1969). Alpha pinene was found not to be a sensitizer for human skin.

Alpha pinene: Maximization test on 25 volunteers (Opdyke DLJ, 1978). The material was tested on humans at a concentration of 10 % in petrolatum and produced no sensitization reactions.

Camphene: Patch test on humans (Woeber K, 1969). Camphene was found not to be a sensitizer for human skin.

Camphene: Maximization test on 25 volunteers (Opdyke DLJ, 1975). The material was tested on humans at a concentration of 4 % in petrolatum and produced no sensitization reactions.

Estragole: Maximization test on 25 volunteers (Opdyke DLJ, 1976). The material was tested on humans at a concentration of 3 % in petrolatum and produced no sensitization reactions.

Based on the experimental data available on the components of the substance and, since content of those classified as skin sensitizers 1B is higher than 1%, it is determined that the substance is classified as skin sensitizer 1B, H317 according to the CLP Regulation (EC) no. 1272/2008.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the available data, the substance needs to be classified for skin sensitization cat. 1B according to CLP Regulation no. 1272/2008.