Schinus molle, ext.

Endpoint summary

Description of key information

Skin sensitisation using read across from Schinus Terebinthifolius(OECD TG 429): Sensitising (extrapolated EC3 of 22.3%)

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 06 to 28, 2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 429 without some restrictions: pooled method was used instead of individual method; and ear thickness measurements were not included. However, these deviations did not affect the outcome of this study since the observed positive evidence of skin sensitisation is coherent with the composition of the UVCB tested.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 24 April 2002

Deviations:

yes

Remarks:

pooled method instead of individual method; no ear thickness measurements

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

national GLP Compliance Programme (inspected on September 02, 2006/ signed on January 19, 2007)

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

CBA/Ca

Remarks:

CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands, B.V. Postbus 6174, NL - 5960 AD Horst / The Netherlands
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 17.5-22 g
- Housing: single housing. Makrolon Type I, with wire mesh top (EHRET GmbH, D-79302 Emmendingen). Granulated soft wood bedding (Harlan Winkelmann GmbH, D-33178 Borchen)
- Diet: pelleted standard diet, ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)
- Water: tap water, ad libitum, (Gemeindewerke, D-64380 Rossdorf)
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature:122 +/- 3°C
- Humidity: 30-70 %
- Air changes: not reported
- Photoperiod: 12 hours continuous light and 12 hours darkness

- IN-LIFE DATES: not reported

Vehicle:

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

Concentration:

10%, 25% or 50% w/w

No. of animals per dose:

4

Details on study design:

PRE-SCREEN TESTS:
- Compound solubility: up to 50% in acetone/olive oil (4:1 v/v)
- Irritation & systemic toxicity: To determine the highest non-irritant test concentration, a pre-test was performed in two animals. Two mice were treated with concentrations of 10, 25, 50, and 100 % on one ear each on three consecutive days. Clinical signs were recorded 24 ± 4 hours after each application. The animal treated with 50 and 100% of the test item died after the second application of the test item. Therefore, a second experiment was performed using test item concentrations of 25% on both ears of one animal and 50 % on both ears of the second animal. At the tested concentrations the animals did not show any signs of irritation or systemic toxicity.
- Ear thickness measurements: not included
- Erythema scores: not reported
The test item in the main study was assayed at 10, 25, and 50%. The top dose is the highest technically achievable concentration whilst avoiding systemic toxicity and excessive local irritation. No severe irritant effects were tolerated choosing the test concentrations.

MAIN STUDY:

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local Lymph Node Assay - pooled method
- Criteria used to consider a positive response: A test item is regarded as a sensitiser in the LLNA if the following criteria are fulfilled:
First, that exposure to at least one concentration of the test item resulted in an incorporation of 3HTdR at least 3-fold or greater than that recorded in control mice, as indicated by the stimulation index.
Second, that the data are compatible with a conventional dose response, although allowance must be made (especially at high topical concentrations) for either local toxicity or immunological suppression.

TREATMENT PREPARATION AND ADMINISTRATION:
The test item was placed into a volumetric flask on a tared balance and acetone:olive oil (4+1) was quantitatively added.
The preparations were made freshly before each dosing occasion.
Concentrations were in terms of material as supplied.
- Topical Application:
Each test group of mice was treated by topical (epidermal) application to the dorsal surface of each ear lobe (left and right) with different test item concentrations of 10, 25,and 50% (w/v) in acetone:olive oil (4+1). The application volume, 25 Ql, was spread over the entire dorsal surface (diameter ca. 8 mm) of each ear lobe once daily for three consecutive days. A further group of mice was treated with an equivalent volume of the relevant vehicle alone (control animals).
- Administration of 3H-Methyl Thymidine:
3H-methyl thymidine (3HTdR) was purchased from GE Healthcare (GE Healthcare product code no. TRA 310; specific activity, 2 Ci/mmol; concentration, 1 mCi/ml). Five days after the first topical application, all mice were administered with 250 Ql of 81.0 QCi/ml 3HTdR (corresponds to 20.25 QCi 3HTdR per mouse) by intravenous injection via a tail vein.
- Determination of Incorporated 3HTdR
Approximately five hours after treatment with 3HTdR all mice were euthanised by intraperitoneal injection of Pentobarbital-Natrium (Release, WDT, D-30827 Garbsen). The draining lymph nodes were rapidly excised and pooled per group (8 nodes per group). Single cell suspensions (in phosphate buffered saline) of pooled lymph node cells were prepared by gentle mechanical disaggregation through stainless steel gauze (200 Qm mesh size). After washing two times with phosphate buffered saline (approx. 10 ml) the lymph node cells were resuspended in 5 % trichloroacetic acid (approx. 1 ml) and incubated at approximately +4 °C for at least 18 hours for precipitation of macromolecules. The precipitates were then resuspended in 5 % trichloroacetic acid (1 ml) and transferred
to plastic scintillation vials with 10 ml of ‘Ultima Gold’ scintillation liquid (Perkin Elmer (LAS) GmbH, D-63110 Rodgau) and thoroughly mixed. The level of 3HTdR incorporation was then measured on a beta-scintillation counter (Tricarb 2900 TR, Perkin Elmer (LAS) GmbH, D-63110 Rodgau). Similarly, background 3HTdR levels were also measured in two 1ml-aliquots of 5 % trichloroacetic acid. The beta-scintillation counter expresses 3HTdR incorporation as the number of radioactive
disintegrations per minute (DPM).

Positive control substance(s):

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

Statistics:

The mean values and standard deviations were calculated.
A statistical analysis was conducted for assessment of the dose-response relationship, and the EC3 value was calculated according to the equation
EC3 = (a-c) [(3-d)/(b-d)] + c
where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining lymph node cell proliferative activity; (a, b) and (c, d) are respectively the co-ordinates of the two pair of data lying immediately above and below the S.I. value of 3 on the local lymph node assay dose response plot.

Positive control results:

The positive control α Hexylcinnamaldehyde gave a Stimulation Index of greater than 3 (4.87) when tested at a concentration of 25% w/v in acetone/olive oil 4:1 and an EC-3 of 15.7 % (w/v) , thus, demonstrating the sensitivity and reliability of the test system.

Key result

Parameter:

SI

Value:

1.62

Test group / Remarks:

10% w/v in acetone/olive oil 4:1

Key result

Parameter:

SI

Value:

3.3

Test group / Remarks:

25% w/v in acetone/olive oil 4:1

Key result

Parameter:

SI

Value:

4.77

Test group / Remarks:

50% w/v in acetone/olive oil 4:1

Key result

Parameter:

EC3

Value:

22.3

Cellular proliferation data / Observations:

DETAILS ON STIMULATION INDEX CALCULATION
Stimulation index for 10, 25 and 50% w/v in acetone/olive oil 4:1 were 1.62, 3.30 and 4.77, respectively.

EC3 CALCULATION
EC3 = (a-c) [(3-d)/(b-d)] + c = 22.3% (w/v)
a = 10
b = 1.62
c = 25
d = 3.30

VIABILITY/MORTALITY/ No deaths occurred during the study period

CLINICAL SIGNS: No symptoms of local toxicity at the ears of the animals and no systemic findings were observed during the main experiment.

BODY WEIGHTS: The body weight of the animals, recorded prior to the first application and prior to treatment with 3HTdR, was within the range commonly recorded for animals of this strain and age.

Table 7.4.1/1: Grouped Disintegrations per Minute and Stimulation Index

Test item concentration % (w/v)	Group	Measurement DPM	Calculation			Result
			DPM-BGa)	number of lymph nodes	DPM per lymph nodeb)	S.I.
---	BG I	27	---	---	---	---
---	BG II	27	---	---	---	---
---	1	6405	6378	8	797.3
10	2	10364	10337	8	1292.1	1.62
25	3	21102	21075	8	2634.4	3.30
50	4	30476	30449	8	3806.1	4.77

BG = Background (1 ml 5% trichloroacetic acid) in duplicate

1 = Control Group

2-4 = Test Group

S.I. = Stimulation Index

a) = The mean value was taken from the figures BG I and BG II

b) = Since the lymph nodes of the animals of a dose group were pooled, DPM/node was determined by dividing the measured value by the number of lymph nodes pooled

Interpretation of results:

other: sensitizer (Category 1B)

Remarks:

in accordance with EU CLP (EC 1272/2008 and its updates)

Conclusions:

Under the test conditions, test material is classified as a contact sensitizer (Category 1B) according to the Regulation (EC) No. 1272/2008 and to the GHS.

Executive summary:

A study was performed to assess the skin sensitisation potential of test material 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. 

In the preliminary screening test, two mice were treated with concentrations of 10, 25, 50, and 100 % w/v on one ear each on three consecutive days. Clinical signs were recorded 24 ± 4 hours after each application. The animal treated with 50 and 100% w/vof the test item died after the second application of the test item. Therefore, a second experiment was performed using test item concentrations of 25% w/v on both ears of one animal and 50 % w/v on both ears of the second animal. At the tested concentrations the animals did not show any signs of irritation or systemic toxicity, therefore, the concentration of 50% w/v was selected as the highest dose investigated in the main test of the Local Lymph Node Assay. Three groups, each of four animals, were treated with 50 µL (25 µL per ear) of the test item as a solution in acetone/olive oil 4:1 at concentrations of 10%, 25% or 50% w/v. A further group of five animals was treated with acetone/olive oil 4:1 alone. The results of a positive control test, using a group of four animals, performed with the known sensitizer, α‑Hexylcinnamaldehyde, at concentrations of 5%, 10% and 25% w/v in acetone/olive oil 4:1 is reported. The proliferative response of the lymph node cells (LNC) from the draining auricular lymph nodes was assessed five days following the initial application, by measurement of the incorporation of 3H-methyl Thymidine (3HTdR) by β-scintillation counting of LNC suspensions. The response was expressed as radioactive disintegrations per minute per pooled lymph node and as the ratio of 3HTdR incorporation into LNC of test nodes relative to that recorded for control nodes (test/control ratio), termed as Stimulation Index (SI).

Stimulation index for 10%, 25% and 50% w/v in acetone/olive oil 4:1 were 1.62, 3.30 and 4.77, respectively. The concentration of test item expected to cause a 3 fold increase in 3HTdR incorporation (extrapolated EC3 value) was calculated to be 22.3 % (w/v). No sign of systemic toxicity or excessive local skin irritation were noted at the concentrations tested.

The positive control α Hexylcinnamaldehyde gave a Stimulation Index of greater than 3 (4.87) when tested at a concentration of 25% w/v in acetone/olive oil 4:1 and an EC-3 of 15.7 % (w/v) , thus, demonstrating the sensitivity and reliability of the test system.

Reference 2

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: read across

Justification for type of information:

The read across justification is presented in the endpoint summary and the accompanying files are also attached there.

Reason / purpose for cross-reference:

read-across source

Key result

Parameter:

SI

Value:

1.62

Test group / Remarks:

10% w/v in acetone/olive oil 4:1

Key result

Parameter:

SI

Value:

3.3

Test group / Remarks:

25% w/v in acetone/olive oil 4:1

Key result

Parameter:

SI

Value:

4.77

Test group / Remarks:

50% w/v in acetone/olive oil 4:1

Key result

Parameter:

EC3

Value:

22.3

Interpretation of results:

other: Skin sensitizer (Category 1B)

Remarks:

in accordance with EU CLP (EC 1272/2008 and its updates)

Conclusions:

Under the test conditions, test material is classified as a contact sensitizer (Category 1B) according to the Regulation (EC) No. 1272/2008 and to the GHS.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

Skin sensitisation (read across from Schinus Terebinthifolius)

The skin sensitisation of Schinus Molle oil was assessed by using read across from Schinus Terebinthifolius (CAS No.: 949495-68-5). First the experimental information of the source substance will be summarised. Thereafter the read across justification is presented. The accompanying files are attached in the present endpoint summary.

 

Local lymph node assay

A study was performed to assess the skin sensitisation potential of test material 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. In the preliminary screening test, two mice were treated with concentrations of 10, 25, 50, and 100 % w/v on one ear each on three consecutive days. Clinical signs were recorded 24 ± 4 hours after each application. The animal treated with 50 and 100% w/v of the test item died after the second application of the test item. Therefore, a second experiment was performed using test item concentrations of 25% w/v on both ears of one animal and 50 % w/v on both ears of the second animal. At the tested concentrations the animals did not show any signs of irritation or systemic toxicity, therefore, the concentration of 50% w/v was selected as the highest dose investigated in the main test of the Local Lymph Node Assay. Three groups, each of four animals, were treated with 50 µL (25 µL per ear) of the test item as a solution in acetone/olive oil 4:1 at concentrations of 10%, 25% or 50% w/v. A further group of five animals was treated with acetone/olive oil 4:1 alone. The results of a positive control test, using a group of four animals, performed with the known sensitizer, α‑Hexylcinnamaldehyde, at concentrations of 5%, 10% and 25% w/v in acetone/olive oil 4:1 is reported. The proliferative response of the lymph node cells (LNC) from the draining auricular lymph nodes was assessed five days following the initial application, by measurement of the incorporation of 3H-methyl Thymidine (3HTdR) by β-scintillation counting of LNC suspensions. The response was expressed as radioactive disintegrations per minute per pooled lymph node and as the ratio of 3HTdR incorporation into LNC of test nodes relative to that recorded for control nodes (test/control ratio), termed as Stimulation Index (SI). Stimulation index for 10%, 25% and 50% w/v in acetone/olive oil 4:1 were 1.62, 3.30 and 4.77, respectively. The concentration of test item expected to cause a 3 fold increase in 3HTdR incorporation (extrapolated EC3 value) was calculated to be 22.3 % (w/v). No sign of systemic toxicity or excessive local skin irritation were noted at the concentrations tested. The positive control α Hexylcinnamaldehyde gave a Stimulation Index of greater than 3 (4.87) when tested at a concentration of 25% w/v in acetone/olive oil 4:1 and an EC-3 of 15.7 % (w/v) , thus, demonstrating the sensitivity and reliability of the test system.

 

Read across justification

Schinus Molle oil (CAS 94334-31-3; target) and its sensitising properties using read across from Essential oil of Schinus Terebinthifolius (Anacardiaceae) obtained from red berries by supercritical carbon dioxide extraction (indicative), hereafter named Pink Pepper oil (CAS 949495-68-5; source)

 

Introduction and hypothesis for the read across

Schinus Molle oil is a UVCB containing hydrocarbon constituents. The main constituents have three C=C double bonds (myrcene), a 6 ring with two C=C double bonds (alpha-phellandrene), a 6 ring with a C=C double bond inside and a C=C double bond outside the ring (DL-Limonene), a 6 ring with a C=C double bond inside and a C=C double bond outside the ring (beta-phellandrene). For Schinus Molle oil (target) no skin sensitisation data are available. Therefore additional information is used in accordance with Article 13 of REACH where it is said that lacking information should be generated whenever possible by means other than vertebrate animal tests, i.e. applying alternative methods such as in vitro tests, SARs, grouping and read-across. For assessing the skin sensitisation of Schinus Molle oil, the sensitisation data of Pink Pepper oil is used.

 

Hypothesis:Schinus Molle oil (target) is expected to have similar sensitising properties based on information from Pink Pepper oil (source) because both have common constituents in comparable concentrations in the oil.

 

Available skin sensitisation information:

The source substance Pink Pepper oil (Essential oil of Schinus Terebinthifolius (Anacardiaceae) obtained from red berries by supercritical carbon dioxide extraction) has been tested in a well performed OECD TG 429 study, under GLP. In the local lymph node assay the test item dissolved in acetone:olive oil (4+1) was assessed for its possible skin sensitising potential using test item concentrations of 10, 25, and 50%. The animals did not show any clinical signs during the course of the main experiment and no cases of mortality were observed. In this study Stimulation Indices (SI) of 1.62, 3.30, and 4.77 were determined with the test item at respective concentrations of 10, 25, and 50%. The test item Pink Pepper oil (ST 08 C 08) was found to be a skin sensitiser as an EC3 value of 22.3 % (w/v) was determined.

 

Target and Source chemical(s):

The composition of the target chemical (Schinus Molle oil) and the source chemical (Pink Pepper oil) are shown in data matrix 1. Schinus Molle oil and Pink Pepper oil are both substances of Unknown or Variable Composition, Complex Reaction Products and Biological Materials (UVCBs), obtained by steam distillation from the Berries from Schinus Molle tree (Anacardiaceae) and by supercritical carbon dioxide extraction (indicative) from the Schinus Terebinthifolius red berries, respectively.

 

Similarities in composition:

The target and the source share nine common constituents: Pin-2(3)-ene, Thuj-4(10)-ene, Pin-2(10)-ene, 7-methyl-3-methyleneocta-1,6-diene, p-mentha-1,5-diene, para-cymene, Dipentene, p-mentha-1(7),2-diene and beta-caryophyllene.

Schinus Molle oil and Pink Pepper oil have a comparable amount of minors/unknowns: 10.00-25.00% w/w present in Schinus Molle oil and 0.01-20.00% w/w present in Pink pepper.

 

Differences in composition:

- p-mentha-1,5-diene (Alpha-phellandrene) and 7-methyl-3-methyleneocta-1,6-diene (myrcene) are present in higher amounts in Schinus Molle oil (target) than in Pink Pepper oil (Source).

- The following two constituents (> 1%) are only present in Schinus Molle oil (target): (+)-Delta cadinene (1.0-5.00%) and Alpha muurolene (0.5-3.00%).

- The following seven constituents (> 1%) are only present in Pink Pepper oil (source): Delta 3-carene (1.00-29.00%), Germacrene D (3.00-14.00%), Germacrene B (0.01-3.00%), Elemol (0.01-6.00%), Spathulenol (0.00-3.00%), Bicyclogermacrene (0.01-3.00%) and Terpinolene (0.01-3.00%).

 

The focus will be on the common constituents that are present in both Schinus Molle oil and Pink Pepper oil, as well as the constituents present in Schinus Molle oil but not in Pink Pepper oil. The information on Schinus Molle oil (target) and the information from Pink Pepper oil (source) are presented in the data matrices below. This includes physico-chemical properties and toxicological information, relevant for sensitisation.

 

Purity / Impurities:

The impurities are not relevant for Schinus Molle oil (target) and Pink Pepper oil (source) as they are both UVCBs, however constituents are applicable. For target and source the constituents are presented in the data matrices below. The similarity and differences in constituents between target and source will be discussed below in the analogue justification.

 

Analogue justification:

According to Annex XI 1.5, read across can be used to replace testing when the similarity between the substances based on composition can be established. When using read across the result derived should be applicable for C&L and/or risk assessment and it should be presented with adequate and reliable documentation.

 

Analogue selection:

Pink Pepper oil was selected as analogue because of the similarity in constituents with Schinus Molle oil (see Data matrix 2). Pink Pepper oil and Schinus Molle oil have many constituents in common in comparable amounts which justifies the analogue approach. The constituents that are present in significant concentrations in Schinus Molle oil but not (or in a lower concentration range) in Pink Pepper oil will be addressed in the Toxico-dynamics section.

 

Bioavailability:Dermal absorption of both UVCBs is expected despite variation in properties of some constituents. The molecular weight and liquid appearance of the constituents in both UVCBs are favorable for dermal absorption. Based on the water solubility and log Kow of all constituents, absorption is anticipated to be low to moderate. However due to the generally high LogKow (>4) of the majority of constituents the rate of penetration may be limited by the rate of transfer between the stratum corneum and the epidermis, but uptake into the stratum corneum will be high.

 

Toxico-dynamics:Reactivity is the key parameter for assessing the skin sensitization potential. Both Schinus Molle and Pink Pepper are expected to have the comparable reactivity based on the similarity in sensitising constituents (a.o. beta caryophyllene, Pin-2(10)-ene (beta pinene) and terpinolene) present in both UVCBs. Two known sensitising constituents (according to the ECHA disseminated dossier) are present in a lower amount in the target than in the source; Pin-2(3)-ene (alpha pinene; 2-8% vs. 8-23%), delta-3-carene (0% vs 1-29%) which could indicate a greater potential for sensitisation of the source UVCB, and thus pose a worst-case situation. Furthermore the concentration range of 7-methyl-3-methyleneocta-1,6-diene (myrcene) is higher in the target substance compared to the source (10-25% vs. 0-5%). Myrcene is reported as Skin Sens. 1 in the classification section of the registration dossier, but did not induce delayed contact hypersensitivity in the murine Local Lymph Node Assay. Based on the test result, no higher sensitising potential is expected in the target substance.

 

Data matrix:

The relevant information on constituents, physico-chemical properties and toxicological characteristics are presented in the data matrix in Table 1 and 2.

 

Uncertainty of the prediction:

Schinus Molle oil (target) and Pink Pepper (source) contain many constituents in common, in comparable concentrations. The constituents of Schinus Molle oil that are not present in Pink Pepper (alpha muurolene and delta cadinene) have similar functional groups as the common constituents, and are not expected to have an effect on the skin sensitising potential. Based on the previous, these constituents are not expected to impair the reliability of this read-across prediction.

 

Conclusions for skin sensitisation:

For Schinus Molle oil skin sensitisation information is not available, and read across from Pink Pepper oil is used. When using read across the result derived should be applicable for C&L and/or risk assessment and be presented with adequate and reliable documentation. The current document presents such documentation. The source substance has been tested in a well conducted OECD TG 429 (LLNA) test, in which it was found to be a skin sensitizer.

 

Final conclusion on hazard and application in the risk characterization: Based on the available data and the read across justification, the target substance Schinus Molle oil is considered a skin sensitiser.

 

Data matrix 1. Information on source and target relevant for assessment of skin sensitisation potential

	Target	Source
Name	Schinus Molle oil	Pink Pepper oil*
Molecular structure	N/A (UVCB)	N/A (UVCB)
CAS	94334-31-3	949495-68-5
REACH registration	To be registered (Annex VII)	REACH registered
EC Number	305-104-2	481-880-7
Molecular formula	N/A (UVCB)	N/A (UVCB)
Molecular weight	N/A	N/A
Physico-chemical properties
Appearance	Liquid	Liquid
Vapour pressure (Pa)	168.9 (Exp)	Range: 2.6 – 372 for 12/13 constituents, one major constituent (alpha-pinene) has a higher value at 851 Pa Range for all constituents: 2.6-851(QSAR and experimental data)
Water solubility (mg/L)	36.9 (Exp)	Range: 0.0007 - 7.14 (10/13 constituents have a water solubility < 1 mg/L) (QSAR and experimental data)
Log Kow	4.7 – 5.7 (Exp)	Range: 3.68 - 6.4, only one minor constituent has <4. (QSAR and experimental data)
Human health
Acute oral tox	LD50 > 5000 mg/kg bw	LD50 > 5000 mg/kg bw
Skin sensitisation	Read-across	Skin Sens. 1B
Skin irritation	Read-across	Not classified
Eye irritation	Not classified	Not classified
Genetic toxicity (Ames)	Not mutagenic	Not mutagenic

Exp =Experimental. Reference: IFF, 2017. The Determination of Physico-Chemical Properties of Schinus Molle oil. Report No. 191244.

QSAR = Quantitative Structure-Activity Relationship

*Physico-chemical, and human health information from Pink Pepper oil are derived from ECHA disseminated dossier (May 2018)

N/A= Not applicable

Data matrix 2. Composition of Target and Source and skin sensitisation information available for these constituents

NAME	CAS	Target	Target	Source	Source	Endpoint specific classification*
		Min.	Max.	Min.	Max.
Pin-2(3)-ene (DL-alpha-pinene)	80-56-8	2.00%	8.00%	8.00%	23.00%	Skin Sens. 1 (based on effects of beta pinene)
Thuj-4(10)-ene (Sabinene)	3387-41-5	2.00%	8.00%	0.01%	5.00%	Not available
Pin-2(10)-ene (Beta-pinene)	127-91-3	0.50%	3.00%	0.01%	3.00%	Skin Sens. 1B
7-methyl-3-methyleneocta-1,6-diene (Myrcene)	123-35-3	10.00%	25.00%	0.01%	5.00%	Skin Sens. 1, but not supported by data: LLNA was negative
p-mentha-1,5-diene (Alpha-phellandrene)	99-83-2	20.00%	35.00%	12.00%	33.00%	Not available
Para-cymene	99-87-6	1.00%	5.00%	0.01%	5.00%	Not classified
Dipentene (Limonene)	138-86-3	5.00%	15.00%	0.01%	15.00%	Skin Sens. 1
p-mentha-1(7),2-diene (Beta-phellandrene)	555-10-2	5.00%	15.00%	0.01%	10.00%	Skin Sens. 1B (EC3 29%)
Caryophyllene (beta-caryophyllene)	13877-93-5/87-44-5	0.50%	4.00%	0.01	6.00%	Skin Sens. 1B
(+)-Delta cadinene	483-76-1	1.00%	5.00%	-	-	Not available
Alpha muurolene	31983-22-9	0.50%	3.00%	-	-	Not available
Delta 3-carene	13466-78-9	-	-	1.00%	29.00%	Skin Sens. 1
Germacrene D	37839-63-7	-	-	3.00%	14.00%	Not available
Germacrene B	15423-57-1	-	-	0.01%	3.00%	Not available
Elemol	639-99-6/8024-27-9	-	-	0.01%	6.00%	Not available
Spathulenol	6750-60-3	-	-	0.00%	3.00%	Not available
Bicyclogermacrene	24703-35-3	-	-	0.01%	3.00%	Not available
Terpinolene	586-62-9	-	-	0.01%	3.00%	Skin Sens. 1B (EC3 10%)
Other minor and unknown constituents	-	10.00%	25.00%	0.01%	20.00%

*Human-health Information from individual constituents is derived from the available ECHA disseminated dossiers (May 2018)

Justification for classification or non-classification

Based on the available information, the substance should be classified as sensitising to the skin (Skin Sens. 1B / H317) in accordance with the criteria outlined in the EU CLP Regulation (1272/2008/EC and its amendments).