Cyclohexadecen-1-one

Endpoint summary

Description of key information

Based on the results of an OECD 404 compliant study, the test item was found to be irritating to rabbit skin.

Based on the results of an OECD 405 compliant study, the test item was found to be not irritating the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003-11-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The in vivo study on skin irritation with the registration substance was completed in 2003. Therefore, in compliance with Regulation (EC) 1907/2006 (REACH) performing an additional in vitro skin irritation study is considered scientifically not necessary.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2001

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

other: Chbb:HM(SPF)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 18 - 22 months
- Weight at study initiation: 2.7 - 3.0 kg
- Housing: individually in PPO cages (floor area: 2576 sq. cm) with perforated floor
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: approx. one week

ENVIRONMENTAL CONDITIONS
- Temperature: 20 +/- 3 °C
- Humidity: 55 +/- 15 %
- Air changes: 10 per hr
- Photoperiod: 12 / 12 hrs dark / hrs light

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: olive oil

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

VEHICLE
- Amount applied: ca. 0.1 mL for a better contact to the skin

Duration of treatment / exposure:

4 h

Observation period:

21 d

Number of animals:

3 females

Details on study design:

TEST SITE
- Area of exposure: back
- Type of wrap: 16-layer gauze patch (2.5 cm x 2.5 cm) was secured semi-occlusively with adhesive Gothaplast tape (2.5 cm wide) and fixed with Gothaplast tape (5 cm wide) loosely wound round the trunk

REMOVAL OF TEST SUBSTANCE
- Washing: with mild soap and lukewarm water
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
The skin reactions were read immediately and 1 h, 24 h, 48 h as well as 72 h after the termination of exposure as well as on day 7, 14 and 21

SCORING SYSTEM:
According to guideline and Draize scoring system:

Erythema and Eschar Formation Score
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema: 4
Maximum possible: 4

Oedema Formation Score
No oedema: 0
Very slight oedema (barely perceptible): 1
Slight oedema (edges of area well defined by definite raising): 2
Moderate oedema (raised approximately 1 mm): 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure): 4
Maximum possible: 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 21 d

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3.67

Max. score:

4

Reversibility:

fully reversible within: 21 d

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

fully reversible within: 21 d

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 21 d

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 21 d

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3.33

Max. score:

4

Reversibility:

fully reversible within: 21 d

Irritant / corrosive response data:

7 days after the termination of exposure isolated scales were observed an the anterior left test field of animal No. 2425 as well as slight crusty scales an the middle and posterior left test field. Animals No. 2461 and No. 2462 showed flat, yellow scales an the whole anterior right test field.

14 days after the termination of exposure isolated, small scales were still recorded an the middle and posterior left test field of animal No. 2425. Animals No. 2461 and No. 2462 still showed isolated scales an the anterior right test fields.

21 days after the termination of exposure animals No. 2425, No. 2461 and No. 2462 were free of any signs of skin irritation.

Other effects:

- Other adverse local effects: none
- Other adverse systemic effects: not observed

Table 1

Animal No	Exposure	Test field	Ersthema formation observed after x hours after termination					Mean (24 -72 h)	Oedema formation after x hours after termination					Mean (24 -72 h)
			0	1	24	48	72		0	1	24	48	72
2425	3 min	Anterior le.	0	1	2	2	2		0	0	0	0	0
	1 h	Median le.	0	1	3	3	3		0	0	2	2	2
	4 h	Posterior le.	1	2	3	3	3	3.00	0	0	2	2	2	2.00
2461	4 h	Anterior le.		2	3	4	4	3.67		2	3	3	3	3.00
2462	4h	Anterior le.		2	4	4	4	4.00		1	4	3	3	3.33

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

Based on the results of this study, the test item was found to be irritating to rabbit skin.

Executive summary:

The skin irritant effect of the test item was investigated according to the method recommended in the OECD Guideline No. 404, "Acute Dermal Irritation/ Corrosion", 2002, and OECD document "Harmonised Integrated Classification System for Human Health and Environmental Hazards of Chemical Substances and Mixtures", 2001, modified. Based on a weight-of-the-evidence analysis of all existing relevant information carried out on the sponsor's own authority the in vivo testing of the test item was considered to be justified. During an initial test using one female albino rabbit a sequential application (exposure time 3 min, 1 h and 4 h) of the test item was carried out on one site of the left back area each. The skin was examined for signs of irritation straight after as well as 1 h, 24 h, 48 h and 72 h after the termination of exposure as well as on day 7, 14 and 21. Since the animal did show very well defined signs of skin Irritation but no full thickness destruction of the skin within the observation period of 21 days it was decided to carry out a confirmatory test with two additional animals in order to allow a categorization of the test item based on the classification criteria (CLP). Two female albino rabbits were exposed to the test item at one skin site on the back during the confirmatory test. After 4-hour exposure time the test item was removed and the skin was examined 1 h, 24 h, 48 h and 72 h after the termination of exposure as well as on day 7, 14 and 21. Skin reactions ranging from well defined to severe were observed in these animals, too. The observation period was extended to the complete reversibility of all effects on day 21 according to the sponsor's request. Under the experimental conditions described in this report the mean scores after the 4-hour exposure were:

Animal No	Erythema	Oedema
2425	3.00	2.00
2461	3.67	3.00
2462	4.00	3.33

 

Under the experimental conditions of this study, the mean score for erythema was 3.6 and for oedema 2.8. Based on the results of this study the test item is considered to be irritating to rabbit skin.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004-12-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The in vivo study on eye irritation/corrosion with the registration substance was completed in 2004. Therefore, in compliance with Regulation (EC) 1907/2006 (REACH) performing an additional in vitro eye irritation/corrosion study is considered scientifically not necessary.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

Himalayan

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: approx. 4.5 months
- Weight at study initiation: 2.1 - 2.2 kg
- Housing: single housing
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 20 d

ENVIRONMENTAL CONDITIONS
- Temperature: 20+/- 3 °C
- Humidity: 30 - 70 %
- Air changes: not specified
- Photoperiod: 12/12 hrs dark / hrs light

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL

Duration of treatment / exposure:

single treatment

Observation period (in vivo):

6 days

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: not done

SCORING SYSTEM: according to the guideline:

CORNEA (Opacity: degree of density (area most dense taken for reading))
0 no ulceration or opacity
1 scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 easily discernible translucent area, details of iris slightly obscured
3 nacreous areas, no details of Iris visible, size of pupil barely discernible
4 opaque cornea, iris not discernible through the opacity

IRIS
0 normal
1 markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection; iris reactive to light (a sluggish reaction is considered to be an effect)
2 haemorrhage, gross destruction, or no reaction to light

CONJUNCTIVAE
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
0 blood vessels normal
1 some blood vessels hyperaemic (injected)
2 diffuse, crimson colour, individual vessels not easily discernible
3 diffuse beefy red

Chemosis: Swelling (refers to lids and/or nictitating membranes)
0 normal
1 some swelling above normal
2 obvious swelling with partial eversion of lids
3 swelling with lids about half-closed
4 swelling with lids more than half-closed

TOOL USED TO ASSESS SCORE: hand-slit lamp

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 24 h

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 24 h

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.66

Max. score:

2

Reversibility:

fully reversible within: 24 h

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 24 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 6 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 6 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 6 days

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 6 days

Table 1 - Eye irritation scores of the right eye of treated animals (animal number 1 / 2 / 3)

Time after administration	Cornea (opacity)	Iris	Conjunctiva (redness)	chemosis
Before dosing	0/0/0	0/0/0	0/0/0	0/0/0
1 h	0/0/0	0/0/0	2/2/2	1/1/1
24 h	1/1/2	1/1/1	2/2/2	1/2/1
48 h	0/0/1	0/0/1	1/2/2	0/2/1
72 h	0/0/0	0/0/0	0/1/1	0/1/1
4 d	-/0/0	-/0/0	-/1/1	-/1/1
5 d	-/0/0	-/0/0	-/1/1	-/1/1
6 d	-/0/0	-/0/0	-/0/0	-/0/0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the test item was irritating the eye.

Executive summary:

A study according to OECD 405 was conducted with the test item. A single instillation of 0.1 mL per animal into the conjuncitval sac of the right eye of three rabbits caused the follwoing changes:

Corneal opacity was observed in all animals:

animal nos. 1 and 2: 24 hours (grade 1) after instillation;

animal no. 3: 24 hours (grade 2) and 48 hours (grade 1) after instillation.

The fluorescein test performed 24 hours after instillation revealed corneal staining in animal nos. 1 and 2 (1/4 of the surface) and animal no. 3 (1/2 of the surface).

Irritation of the iris (grade 1) was observed in all animals 24 hours, in animal no. 3 until 48 hours after instillation.

Conjunctival redness (grade 1 or 2) was observed in all animals 60 minutes to 48 hours, in animal nos. 2 and 3 until 5 days after instillation.

Conjunctival chemosis (grade 1 or 2) was observed in all animals 60 minutes to 24 hours, in animal nos. 2 and 3 until 5 days after instillation.

There were no systemic intolerance reactions.

Based on the results, the test item is considered to be not irritating to the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

An in vitro skin irritation and eye irritation/corrosion study were not conducted because adequate data from in vivo testing are available.

Skin irritation in vivo, OECD 404

The skin irritant effect of the test item was investigated according to the method recommended in the OECD Guideline No. 404, "Acute Dermal Irritation/ Corrosion", 2002, and OECD document "Harmonised Integrated Classification System for Human Health and Environmental Hazards of Chemical Substances and Mixtures", 2001, modified. Based on a weight-of-the-evidence analysis of all existing relevant information carried out on the sponsor's own authority the in vivo testing of the test item was considered to be justified. During an initial test using one female albino rabbit a sequential application (exposure time 3 min, 1 h and 4 h) of the test item was carried out on one site of the left back area each. The skin was examined for signs of irritation straight after as well as 1 h, 24 h, 48 h and 72 h after the termination of exposure as well as on day 7, 14 and 21. Since the animal did show very well defined signs of skin Irritation but no full thickness destruction of the skin within the observation period of 21 days it was decided to carry out a confirmatory test with two additional animals in order to allow a categorization of the test item based on the classification criteria (CLP). Two female albino rabbits were exposed to the test item at one skin site on the back during the confirmatory test. After 4-hour exposure time the test item was removed and the skin was examined 1 h, 24 h, 48 h and 72 h after the termination of exposure as well as on day 7, 14 and 21. Skin reactions ranging from well defined to severe were observed in these animals, too. The observation period was extended to the complete reversibility of all effects on day 21 according to the sponsor's request. Under the experimental conditions described in this report the mean scores after the 4-hour exposure were:

Animal No	Erythema	Oedema
1	3.00	2.00
2	3.67	3.00
3	4.00	3.33

 

Under the experimental conditions of this study, the mean score for erythema was 3.6 and for oedema 2.8. Based on the results of this study the test item is considered to be irritating to rabbit skin.

Eye irritation, in vivo, OECD 405

A study according to OECD 405 was conducted with the test item. A single instillation of 0.1 mL per animal into the conjuncitval sac of the right eye of three rabbits caused the follwoing changes:

Corneal opacity was observed in all animals:

animal nos. 1 and 2: 24 hours (grade 1) after instillation; animal no. 3: 24 hours (grade 2) and 48 hours (grade 1) after instillation.

The fluorescein test performed 24 hours after instillation revealed corneal staining in animal nos. 1 and 2 (1/4 of the surface) and animal no. 3 (1/2 of the surface).

Irritation of the iris (grade 1) was observed in all animals 24 hours, in animal no. 3 until 48 hours after instillation.

Conjunctival redness (grade 1 or 2) was observed in all animals 60 minutes to 48 hours, in animal nos. 2 and 3 until 5 days after instillation.

Conjunctival chemosis (grade 1 or 2) was observed in all animals 60 minutes to 24 hours, in animal nos. 2 and 3 until 5 days after instillation.

There were no systemic intolerance reactions.

Based on the results, the test item is considered to be not irritating to the eye.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data, the test item is classified and labelled as irritating to the skin Cat. 2 (H315: "Causes skin irritation") according to Regulation (EC) No 1272/2008 (CLP), as amended for the fourteenth time in Regulation (EU) 2020/217. The test item does not require classification as irritating to the eye according to CLP.