Eucalyptus globulus, ext.

Endpoint summary

Description of key information

- Skin irritation/corrosion: irritating (WoE).
- Eye irritation: not irritating (OECD 405, GLP, K, rel.1).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1973

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Only short abstract available

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Skin irritation was assessed in an acute dermal toxicity study.

GLP compliance:

no

Remarks:

pre-GLP

Species:

rabbit

Strain:

not specified

Details on test animals or test system and environmental conditions:

None

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

not specified

Controls:

no

Amount / concentration applied:

5000 mg/kg bw

Duration of treatment / exposure:

No data

Observation period:

14 days

Number of animals:

10

Details on study design:

Animals were observed for mortality, clinical signs of toxicity and dermal reactions for 14 days.

Irritation parameter:

erythema score

Basis:

other: 5/10 animals

Remarks on result:

other: slight redness in 5/10 rabbits

Irritation parameter:

erythema score

Basis:

other: 3/10 animals

Remarks on result:

other: moderate redness in 3/10 rabbits

Irritation parameter:

edema score

Basis:

other: all animals

Remarks on result:

other: moderate oedema in 10/10 rabbits

Irritant / corrosive response data:

Slight redness (5/10 rabbits), moderate redness (3/10 rabbits) and moderate oedema (10/10 rabbits).

Other effects:

No mortality was observed.

None

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

Dermal reactions noted were slight redness (5/10 rabbits), moderate redness (3/10 rabbits) and moderate oedema (10/10 rabbits) at the site of application.

Executive summary:

In an acute dermal toxicity study, 10 rabbits were administered a single dermal dose of eucalyptus oil at 5000 mg/kg bw. Animals were then observed for mortality and clinical signs of toxicity for 14 days.

Dermal reactions noted were slight redness (5/10 rabbits), moderate redness (3/10 rabbits) and moderate oedema (10/10 rabbits) at the site of application.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1998

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Data on constituents only

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Method: Open mouse ear assay (Evans and Schmidt, 1979)

GLP compliance:

not specified

Species:

mouse

Strain:

other: albino

Type of coverage:

open

Preparation of test site:

not specified

Vehicle:

other: acetone

Controls:

no

Duration of treatment / exposure:

No data

Observation period:

24 and 48 h after exposure

Number of animals:

10

Irritation parameter:

other: ID50 (Irritant dose in 50 % individuals)

Basis:

mean

Time point:

other: 48 h

Score:

0.351 - 2.483

Reversibility:

no data

Other effects:

The ID50 values for 1,8-cineole, β-caryophyllene, citronellal, α-terpineol, p-cymene, cryptone were calculated to be 1.008, 2.483, 0.351, 0.706, 0.847 and 0.497 µg/5 µL, respectively.

None

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

Under the test conditions, eucalyptus oil constituents were considered as possibly harmful and irritant to the skin.

Executive summary:

In a study, eucalyptus oil was obtained from the leaves of Eucalyptus globulus (source: waste and uncultivated areas of Lahore, Pakistan) and purified to isolate its active compounds: 1,8-cineole, β-caryophyllene, citronellal, α-terpineol, p-cymene, cryptone (9.39 %) and other minor constituents. Each fraction (10 mg) was dissolved in acetone (10 mL) and applied to the ear of groups of albino mice (10/dose) at the dose levels of 0.078125-20 µg/5 µL. The total number of red ears per dilution and ID50 (Irritant dose in 50 % individuals) were calculated by probit analysis.

The ID50 values for 1,8-cineole, β-caryophyllene, citronellal, α-terpineol, p-cymene, cryptone were calculated to be 1.008, 2.483, 0.351, 0.706, 0.847 and 0.497 µg/5 µL, respectively.

Under the test conditions, eucalyptus oil constituents were considered as possibly harmful and irritant to the skin.

Reference 3

Endpoint:

skin irritation / corrosion, other

Remarks:

Classification based on calculation rules for mixtures of the CLP Regulation

Type of information:

calculation (if not (Q)SAR)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

accepted calculation method

Qualifier:

no guideline required

Principles of method if other than guideline:

Classification based on calculation rules for mixtures of the CLP Regulation

Irritation parameter:

other: classification

Remarks on result:

other: skin irritant category 2

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Executive summary:

The NCS is composed of several identified constituents and in that, it can be considered as a mixture according to the definition of the CLP Regulation.

The decision logic for classification of mixtures from the ECHA Guidance on the Application of the CLP Criteria (2015) was used to determine the skin irritation/corrosion hazard of the registered substance. The decision of classification as skin irritant was based on existing data on constituents (additivity principles): the registered substance has more than 10% of its constituents classified as Skin irritant Category 2 and should be classified as a skin irritant without further testing according to the rules for classification of mixtures of Regulation (EC) No 1272/2008

Constituent	CAS	Classification	Source
Pinene alpha	80 -56 -8	H315	ECHA C&L inventory–self classification
Pinene beta	127 -91 -3	H315	ECHA C&L inventory - self classification
Myrcene beta	123 -35 -3	H315	ECHA C&L inventory - self classification
dipentene	138 -86 -3	H315	Harmonised classification Annex VI of Regulation (EC) No 1272/2008 (CLP Regulation)
alpha-terpineol	98 -55 -5	H315	ECHA C&L inventory - self classification

Source: ECHA disseminated dossiers or self classification

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2013-03-18 to 2013-03-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to OECD test guideline No. 405 and in compliance with GLP.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

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)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS: New Zealand White (Hsdlf:NZW) strain rabbit
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: twelve to twenty weeks old.
- Weight at study initiation: 2.38 to 2.95 kg.
- Housing: individually housed in suspended cages.
- Diet (e.g. ad libitum): free access to food (2930C Teklad Global Certified Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK)
- Water (e.g. ad libitum): free access to drinking water.
The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
- Acclimation period: at least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23
- Humidity (%): 30-70
Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study.
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: the left eye of each rabbit served as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

The eyes were not rinsed after administration of the test item.

Observation period (in vivo):

approximately 1 hour and 24, 48 and 72 hours following treatment

Number of animals or in vitro replicates:

3

Details on study design:

ANALGESIA
Sixty to seventy minutes prior to administration of the test item, buprenorphine 0.01 mg/kg was administered by subcutaneous injection (SC) to provide a therapeutic level of systemic analgesia.

LOCAL ANESTHESIC
Nineteen and five minutes prior to administration of the test item, a local anesthetic (0.5% tretracaine hydrochloride) was applied to each eye.

PAIN MANAGEMENT
Eight hours following the test item administration, buprenorphine 0.01 mg/kg SC and meloxicam 0.5 mg/kg SC was administered to provide a therapeutic level of systemic analgesia. The treated animals were checked for signs of pain and suffering approximately twelve hours later. No further analgesia was required.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The eyes were not rinsed after administration of the test item.

SCORING SYSTEM: Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49.

TOOL USED TO ASSESS SCORE: light source from a standard ophthalmoscope.

OTHER
Any clinical signs of toxicity, if present, were also recorded.
Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2 & #3

Time point:

other: Mean 24-48-72 hours

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2 & #3

Time point:

other: Mean 24-48-72 hours

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: #1 & #3

Time point:

other: Mean 24-48-72 hours

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: Mean 24-48-72 hours

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal: #1, #2 & #3

Time point:

other: Mean 24-48-72 hours

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

No corneal or iridial effects were noted during the study. Moderate conjunctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24 and 48-Hour observations. All treated eyes appeared normal at the 72-Hour observation.

Other effects:

One animal showed no gain in body weight and two animals showed expected gain in body weight during the study.

Table 7.3.2/1: Irritant/corrosive response data each animals at each observation time up to removal from the test

 

Score at time point / Reversibility	Cornea		Iris (/2)	Conjunctivae
	Opacity (/4)	Area (/4)		Redness (/3)	Chemosis (/4)	Discharge (/3)
1 h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	2 / 2 / 2	2 / 2 / 2	2 / 2 / 2
24 h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	1 / 2 / 1	1 / 1 / 1	0 / 0 / 0
48 h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	1 / 1 / 1	1 / 1 / 1	0 / 0 / 0
72 h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0
Average 24h, 48h, 72h	0 / 0 / 0	0 / 0 / 0	0 / 0 / 0	0.7 / 1 / 0.7	0.7 / 0.7 / 0.7	0 / 0 / 0
Reversibility*)	-	-	-	c.	c.	c.
Average time (unit) for reversion	-	-	-	72 h	72 h	24 h

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0/0.0/0.0 for cornea opacity, 0.0/0.0/0.0 for iris lesions, 0.7/1.0/0.7 for redness of the conjunctivae and 0.7/0.7/0.7 for chemosis.

Executive summary:

In an eye irritation study performed according to the OECD guideline No. 405, and in compliance with GLP, 0.1 mL of undiluted Eucalyptus globulus oil was instilled initially into the right eye of a single New Zealand White rabbit. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated. The eyes were not rinsed after administration of Eucalyptus globulus oil. The left eye of each rabbit served as control. Animals were observed 1, 24, 48 and 72 hours after dosing under a light source from a standard ophthalmoscope. The reactions in the conjunctiva (redness, chemosis and discharge), the iris and the cornea (opacity and area involved) were scored according to the Draize scale. 

No corneal or iridial effects were noted during the study. Moderate conjunctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24 and 48-Hour observations. All treated eyes appeared normal at the 72-Hour observation.

One animal showed no gain in body weight and two animals showed expected gain in body weight during the study.

Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0/0.0/0.0 for cornea opacity, 0.0/0.0/0.0 for iris lesions, 0.7/1.0/0.7 for redness of the conjunctivae and 0.7/0.7/0.7 for chemosis.

Under the test conditions, Eucaltus globulus oil is not classified as irritating to eyes according to the criteria of the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and of the Directive 67/548/EEC.

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

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

Justification for type of information:

In accordance with section 1.1 (Use of existing data) of Annex XI to the REACH Regulation, the in-vitro eye irritation study (required in section 8.2) does not need to be conducted as a reliable in-vivo study is available and provides sound conclusive evidence that the substance is non-irritating.

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation / corrosion:

A weight-of-evidence analysis, taking all existing and relevant data, was used to conclude on the skin irritation potential of Eucalyptus oil. The decision of classification as skin irritant was based on the following arguments:

1 - Existing human data: According to Saeed and Sahbir (1998), irritation due to Eucalyptus oil was observed in the local people, who deals with the manufacture of cosmetics and liniments, where this oil was used as their main ingredient. The composition of the Eucalyptus oil tested is not similar to the submitted dossier, therefore the data by itself was not considered sufficiently adequate to conclude on classification.

2 - Existing animal data from irritation study: Skin irritation was observed in a reliable acute dermal toxicity study (Moreno, 1973) at the dose of 5000 mg/kg bw. Dermal reactions noted were slight redness (5/10 rabbits), moderate redness (3/10 rabbits) and moderate oedema (10/10 rabbits) at the site of application. Adequate scoring of skin effects was not provided, therefore it was not possible to take a decision of classification based solely on this study.

3 - Existing data on constituents: As detailed on the publication of Saeed and Sahbir, irritant effect of Eucalyptus oil is mainly due to the monoterpenes present in oil. Indeed, more than 10% of the Eucalyptus oil constituents are classified as skin irritant (i.e. pinene, terpineol). Therefore according to the Regulation (EC) No 1272/2008, the mixture should be classified.

4- Classification of constituents: The NCS is composed of several identified constituents and in that, it can be considered as a mixture according to the definition of the CLP regulation. Alpha pinene, Limonene, Terpineol alpha, Beta myrcene and beta pinene, are all classified as Skin irritant (self or harmonised classification) and are presents above the CLP generic concentration limit of 10%. Therefore, the registered substance is classified as skin irritant according to the Regulation (EC) No 1272/2008.

Eye irritation:

Eucalyptus oil is not a skin corrosive and no damage to the eyes was reported in existing in vitro and/or in vivo studies on more than 90% of Eucalyptus oil constituents. Based on the whole data available, an in vitro test to assess corrosivity was not justified. However, based on the complexity of the substance and on the absence on in vivo data on its major constituent, a new in vivo test was performed.

This in vivo eye irritation study was performed according to the OECD guideline No. 405, and in compliance with GLP (Pooles, 2013). No corneal or iridial effects were noted during the study. Moderate conjunctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24 and 48-Hour observations. All treated eyes appeared normal at the 72-Hour observation. One animal showed no gain in body weight and two animals showed expected gain in body weight during the study. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0/0.0/0.0 for cornea opacity, 0.0/0.0/0.0 for iris lesions, 0.7/1.0/0.7 for redness of the conjunctivae and 0.7/0.7/0.7 for chemosis.

Under the test conditions, Eucalyptus globulus oil is not classified as irritating to eyes

Justification for selection of skin irritation / corrosion endpoint:
Available data indicates that the criteria are met for classification as irritating to the skin.

Justification for selection of eye irritation endpoint:
Only one study available, GLP-compliant and of high quality (Klimisch score = 1).

Effects on skin irritation/corrosion: irritating

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008 including ATP3.

Self-classification:

Based on the available information the substance is classified as:

- Skin Irritant Category 2 (H315: Causes skin irritation) according to the criteria of the Annex VI of the Regulation (EC) No. 1272/2008 (CLP).

No additional self-classification is proposed regarding eye irritation according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and of the Directive 67/548/EC. No information was available regarding respiratory irritation.