Ferula galbaniflua, ext.

Endpoint summary

Description of key information

- Skin irritation/corrosion: irritating, based on the rules of the CLP Regulation for classification of mixtures 
- Eye irritation: not irritating (read-across, OECD TG 437, GLP, Rel.1)

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 January 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well conducted and well described study in accordance with GLP and OECD Guideline 437 without any deviation.

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 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP Compliance Programme (inspected on 10 July 2012/ signed on 30 November 2012)

Species:

other: bovine eye

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST SYSTEM
- Eyes from adult cattle were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised and placed in Hanks' Balanced Salt Solution (HBSS), supplemented with Penicillin/Streptomycin, and transported to the laboratory on ice packs. The corneas were prepared immediately on arrival.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL was applied on each cornea
- Concentration: Test material: undiluted; negative control: 0.9 % w/v sodium chloride solution; positive control: ethanol

Duration of treatment / exposure:

The undiluted test item was applied for 10 minutes at 32 ± 1 °C followed by an incubation period of 120 ± 10 minutes at 32 ± 1 °C.

Observation period (in vivo):

- Corneal opacity was measured pre-treatment, post-treatment and post-incubation (after 120 ± 10 minutes of incubation).
- Application of Sodium Fluorescein (4 mg/mL) and corneal permeability was measured after 90 min of incubation at 32 ± 1 °C.

Number of animals or in vitro replicates:

Total: 9 corneas - 3 corneas/group for test item, negative and positive controls

Details on study design:

Details of test procedure: Corneas obtained from freshly slaughtered adult cattle (from a local abattoir) were mounted in corneal holders. Both chambers of each BCOP holder were filled with complete MEM culture media (MEM) and incubated for 60 minutes at 32 ± 1 °C. Before the treatment, opacity measurement was performed using an opacitometer. The test item (0.75 mL) was applied on each cornea for 10 minutes at 32 ± 1 °C. At the end of the exposure period the test item and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete MEM. The anterior chamber was refilled with fresh complete MEM. A post-treatment opacity reading was taken and each cornea was visually observed. The holders were incubated, anterior chamber facing forward, at 32 ± 1 °C for 120 ± 10 minutes. After incubation the holders were removed from the incubator and a final opacity reading was taken. Each cornea was visually observed. Following the opacity measurement the permeability of the corneas to sodium fluorescein was evaluated. The medium from the anterior chamber was removed and replaced with 1 mL of sodium fluorescein solution (4 mg/mL). The dosing holes were plugged and the holders incubated, anterior chamber uppermost, at 32 ± 1 °C for 90 minutes. After incubation the medium in the posterior chamber of each holder was decanted and retained. 360 µL of medium representing each cornea was applied to a designated well on a 96-well plate and the optical density at 492 nm (OD492) was measured.

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

2.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

IVIS = 0.3

Positive controls validity:

valid

Remarks:

IVIS = 50.5

Irritant / corrosive response data:

- In Vitro Irritancy Score (IVIS) were 2.8, 0.3 and 50.5 for test item, negative and positive controls, respectively.

Other effects:

- The corneas treated with the test item were clear post treatment and post incubation.
- The corneas treated with the negative control item were clear post treatment and post incubation.
- The corneas treated with the positive control item were cloudy post treatment and post incubation.

See the attached document for information on tables of results

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions, Pinus Pinaster Oil, induced an IVIS of 2.8 and is not classified as corrosive or severe irritant for the isolated bovine cornea. Furthermore as the IVIS score is lower than 3, it can be concluded that the test item is not an ocular irritant at all.

Executive summary:

In an in vitro eye irritation study performed according to the OECD Guideline 437 and in compliance with GLP, 0.75 mL of Pinus Pinaster Oil was applied to isolated bovine corneas mounted in corneal holders, and incubated for 120 ± 10 minutes at 32 ± 1 °C using the closed chamber method. Three corneas were used for each treated series (undiluted test item; negative control; positive control: ethanol). Before the treatment, a first opacity measurement was performed using an opacitometer. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). 

The test item, negative and positive control induced an IVIS of 2.8, 0.3 and 50.5, respectively. The corneas treated with the negative control and test item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation. The positive control In Vitro Irritancy Score was within the range of 30.9 to 67.7, therefore the acceptance criterion was satisfied.

 

Under the test conditions, Pinus Pinaster Oil, induced an IVIS of 2.8 and is not classified as corrosive or severe irritant for the isolated bovine cornea. Furthermore as the IVIS score is lower than 3, it can be concluded that the test item is not an ocular irritant and therefore Pinus pinaster oil is not classified for eye irritation according to the Regulation (EC) No. 1272/2008 and the Directive 67/548/EEC.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

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

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

2.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

IVIS = 0.3

Positive controls validity:

valid

Remarks:

IVIS = 50.5

Irritant / corrosive response data:

- In Vitro Irritancy Score (IVIS) were 2.8, 0.3 and 50.5 for test item, negative and vehicle controls, respectively.

Other effects:

- The corneas treated with the test item were clear post treatment and post incubation.
- The corneas treated with the negative control item were clear post treatment and post incubation.
- The corneas treated with the positive control item were cloudy post treatment and post incubation.

See the attached document for information on tables of results

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, Pinus Pinaster Oil, induced an IVIS of 2.8 and is not classified as corrosive or severe irritant for the isolated bovine cornea. Furthermore as the IVIS score is lower than 3, it can be concluded that the test item is not an ocular irritant at all. Therefore, the registered substance Galbanum oil is not an occular irritant.

Executive summary:

In an in vitro eye irritation study performed according to the OECD Guideline 437 and in compliance with GLP, 0.75 mL of Pinus Pinaster Oil was applied to isolated bovine corneas mounted in corneal holders, and incubated for 120 ± 10 minutes at 32 ± 1 °C using the closed chamber method. Three corneas were used for each treated series (undiluted test item; negative control; positive control: ethanol). Before the treatment, a first opacity measurement was performed using an opacitometer. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). 

The test item, negative and positive control induced an IVIS of 2.8, 0.3 and 50.5, respectively. The corneas treated with the negative control and test item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation. The positive control In Vitro Irritancy Score was within the range of 30.9 to 67.7, therefore the acceptance criterion was satisfied.

 

Under the test conditions, Pinus Pinaster Oil, induced an IVIS of 2.8 and is not classified as corrosive or severe irritant for the isolated bovine cornea. Furthermore as the IVIS score is lower than 3, it can be concluded that the test item is not an ocular irritant and therefore the registered substance Galbanum oil is not classified for eye irritation according to the Regulation (EC) No. 1272/2008 and the Directive 67/548/EEC.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion:

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	Classification according to the Regulation (EC) No. 1272/2008 (CLP)		Source
	Skin irritation	Eye irritation
alpha-pinene	SCI 2 (H315)	-	Self-classification
beta-pinene	SCI 2 (H315)	-	Self-classification
delta-3-carene	SCI 2 (H315)	-	Self-classification

Source: ECHA disseminated dossiers

Eye irritation:

In an in vitro eye irritation study performed according to the OECD Guideline 437 and in compliance with GLP, 0.75 mL of Pinus Pinaster Oil was applied to isolated bovine corneas mounted in corneal holders, and incubated for 120 ± 10 minutes at 32 ± 1 °C using the closed chamber method. Three corneas were used for each treated series (undiluted test item; negative control; positive control: ethanol). Before the treatment, a first opacity measurement was performed using an opacitometer. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). 

The test item, negative and positive control induced an IVIS of 2.8, 0.3 and 50.5, respectively. The corneas treated with the negative control and test item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation. The positive control In Vitro Irritancy Score was within the range of 30.9 to 67.7, therefore the acceptance criterion was satisfied.

 

Under the test conditions, Pinus Pinaster Oil, induced an IVIS of 2.8 and is not classified as corrosive or severe irritant for the isolated bovine cornea.Furthermore as the IVIS score is lower than 3, it can be concluded that the test item is not an ocular irritant and thereforethe registered substance Galbanum oil is not classified for eye irritation according to the Regulation (EC) No. 1272/2008 and the Directive 67/548/EEC.

Justification for classification or non-classification

Harmonized classification:

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

Self-classification:

Based on the available information and typical composition provided by the Lead Registrant, the registered substance is classified as skin irritant: Skin Irritant Category 2 (H315: Causes skin irritation) according to the criteria of the Regulation (EC) No. 1272/2008 (CLP).

Based on the available data on Pinus pinaster oil, the registered substance Galbanum oil is not classified for eye irritation according to the criteria of the Regulation (EC) No. 1272/2008 (CLP) .

No information was available regarding respiratory irritation.