(1'R,2R)-2-Methyl-4-(2',2',3'-tri-nal and methylcyclopent-3'-en-1'-yl)-4-pent-enal

Endpoint summary

Description of key information

Skin irritation, in vivo (rabbit): non-irritating, EU Method B.4, 1995

Eye irritation, in vivo (rabbit): non-eye irritating, EU Method B.5, 1995

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:

29-11-1994 to 03-01-1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met.

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: Adult
- Weight at study initiation: 4.3 – 4.9 kg
- Housing: Individually housed in grid bottomed metal cages
- Diet: certified rabbit diet ad libitum (details in the full study report)
- Water: filtered certified water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 -21
- Humidity (%): 47 - 67
- Air changes (per hr): Not reported, however reported as air conditioned
- Photoperiod: 12 hours light / 12 hours dark

IN-LIFE DATES: From: 29-11-1994 To: 14-12-1994

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5mL
- Concentration (if solution): Not applicable.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours (initial observation); additional observations are made daily up to and including on Days 7 and 15 to assess the reversibility of skin reactions (as appropriate).

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: dorsal
- Type of wrap if used: semi-occlusive ; gauze patch secured with semi-occlusive dressing and restraining bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, gently cleansed using cotton wool soaked in warm water.
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
1, 24, 48, 72 hours (initial observations); additional observations are made daily up to and including on Days 7 and 15 to assess the reversibility of skin reactions (as appropriate).

SCORING SYSTEM:
Consistent with Draize. Erythema and Eschar Formation:
No erythema ____________________________0
Very slight erythema (barely perceptible) _____1
Well-defined erythema ____________________2
Moderate to severe erythema ______________3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) _____________4

Oedema Formation
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 millimetre) _________________________________3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) __4
Any other skin reactions and clinical signs of toxicity (clinical signs/behaviour), if present, were also recorded.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 1h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 1h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 1h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 1h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

- Erythema: Slight or well defined erythema (score = 1 or 2) at all treated skin sites at 24 to 72 hours after patch removal and up to day 7. All effects reversed by day 14.
- Edema: Slight or moderated edema (score = 0, 1 or 2) was noted during the study and up to day 7. All effects reversed by day 14.
- Reversibility of effects: All effects reversed within 14 days (3/3 sites).

Other effects:

- Other adverse local effects: None reported
- Other adverse systemic effects: None reported.

Table 1. Individual skin reactions

Skin Reaction	Observation Time	Individual Scores
		1 (female)	2 (female)	3 (female)
Erythema/Eschar formation	1 hour	0	0	1
	24 hours	1	2	1
	48 hours	2	2	1
	72 hours	2	2	2
	7 days	2	2	1
	14 days	0	0	0
Oedema formation	1 hour	1	0	0
	24 hours	1	0	2
	48 hours	1	1	1
	72 hours	1	1	1
	7 days	1	1	1
	14 days	0	0	0

 

Mean scores per organism at 24, 48 and 72h:

Erythemea/Escar Formation:

1: total = 5; mean score = 1.7

2: total = 6; mean score = 2.0

3: total = 4; mean score = 1.3

Oedema Formation:

1: total = 3; mean score = 1.0

2. total = 2; mean score = 0.7

3. total = 4; mean score = 1.3

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the test item is not considered to be irritating.

Executive summary:

The study was performed to EU Method B.4 and/or a method equivalent or similar to OECD TG 404 under GLP to assess the primary skin irritancy potential of the test item in New Zealand White rabbits. Following single 4-Hour, semi-occluded applications to the intact rabbit skin. 0.5 mL of the test item was introduced under a square of surgical lint and placed in position on the clipped skin via semi-occlusive dressing to assess the irritancy potential of the test item. The surgical lint was secured in position with a strip of adhesive bandage. After 4 hours of exposure to the test item, the patches were removed, and individual dose sites were scored at approximately 1, 24, 48, and 72 hours. Daily observations were made up to day 14 as applicable. A single 4-Hour, semi occluded application of the test item to the intact skin of three rabbits produced slight or well-defined erythema (score = 1 or 2) and edema (score = 0, 1 or 2) at all treated skin sites at 24 to 72 hours after patch removal and up to day 7. Mean scores following grading at 24, 48 and 72h were 1.7, 2.0 and 1.3 in erythema and eschar and 1.0, 0.7 and 1.3 for the edema scoring criteria. All effects reversed by day 14. Under the conditions of the study, the test item is not considered to be a skin irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not 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:

10-01-1995 to 27-01-1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: Young adult
- Weight at study initiation: 2.6 – 2.9 kg
- Housing: Individually housed in grid bottomed metal cages
- Diet: certified rabbit diet ad libitum (details in the full study report)
- Water: filtered certified water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 -21
- Humidity (%): 35 - 60
- Air changes (per hr): Not reported, however reported as air conditioned
- Photoperiod: 12 hours light / 12 hours dark

IN-LIFE DATES: From: 10-01-1995 To: 14-01-1995

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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

Duration of treatment / exposure:

A volume of 0.1 mL of the test item, was placed into the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for a few seconds immediately and then moved gently to distribute the test item around the surface of the eye. The left eye remained untreated (and was used for control purposes).

Observation period (in vivo):

Ocular assessment was conducted at approximately 1, 24, 48 and 72 hours after instillation of the test item, according to numerical evaluation.

Number of animals or in vitro replicates:

3 (sex not specified).

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): None
- Time after start of exposure: Not applicable.

SCORING SYSTEM:
The irritation was assessed using criteria consistent with Draize (1977) numerical scoring system. At each observation period, each eye was assessed for damage or irritation to the cornea, iris and conjunctiva using the untreated eye as a control.

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light from a standard light source according to BS 950 (artifical daylight for the assessment of colour).

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: mean; n=3

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: mean; n=3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

other: mean; n=3

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: mean; n=3

Irritant / corrosive response data:

No corneal or Iridial inflammation effects were noted. Very slight conjunctival reactions (redness, score = 1) was noted in treated eyes 1 hour after treatment. No chemosis effects were noted. All effects resolved within 24 hours observation.

Other effects:

- Lesions and clinical observations: None reported.
- Ophthalmoscopic findings: None reported.
- Histopathological findings: None reported.
- Effects of rinsing or washing: Not applicable.
- Other observations: None reported.

Table 1. Individual scores and mean scores for 24, 48 and 72 hours

Organism number and sex	Time after treatment	Corneal Opacity	Iridial Inflammation	Conjunctival Redness	Conjunctival Chemosis
1 (sex not specified)	1h	0	0	1	0
	24 h	0	0	0	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Day 4	-	-	-	-
Mean (24 – 72 h)		0.00	0.00	0.00	0.00
2 (sex not specified)	1h	0	0	1	0
	24 h	0	0	0	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Day 4	-	-	-	-
Mean (24 – 72 h)		0.00	0.00	0.00	0.00
3 (sex not specified)	1h	0	0	1	0
	24 h	0	0	0	0
	48 h	0	0	0	0
	72 h	0	0	0	0
	Day 4	0	0	0	0
Mean (24 – 72 h)		0.00	0.00	0.00	0.00

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the test item is not irritating to the eye.

Executive summary:

The study was performed to EU Method B.5 and/or a method equivalent or similar to OECD TG 405 under GLP to assess the irritancy potential of the test item to the eye following a single application in the New Zealand White rabbit. A volume of 0.1 ml of the test item was placed into the left eye of three animals sequentially using the first as a sentinel. The other eye remained untreated and was used for control purposes. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. A single application of the test item to the non-irrigated eye of three rabbits produced no corneal or Iridial inflammation effects. Slight conjunctival reaction (hyperaemia or redness, score = 1 and/or discharge, score = 1) was noted in treated eyes at 1 hour after treatment. No chemosis effects were noted. All treated eyes appeared normal at the 24 hours observation point. Under the conditions of this study, the test item is not considered to be 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

Skin Irritation:

Key study : In vivo, EU Method B.4, 1995 : The study was performed to EU Method B.4 and/or a method equivalent or similar to OECD TG 404 under GLP to assess the primary skin irritancy potential of the test item in New Zealand White rabbits. Following single 4-Hour, semi-occluded applications to the intact rabbit skin. 0.5 mL of the test item was introduced under a square of surgical lint and placed in position on the clipped skin via semi-occlusive dressing to assess the irritancy potential of the test item. The surgical lint was secured in position with a strip of adhesive bandage. After 4 hours of exposure to the test item, the patches were removed, and individual dose sites were scored at approximately 1, 24, 48, and 72 hours. Daily observations were made up to day 14 as applicable. A single 4-Hour, semi occluded application of the test item to the intact skin of three rabbits produced slight or well-defined erythema (score = 1 or 2) and edema (score = 0, 1 or 2) at all treated skin sites at 24 to 72 hours after patch removal and up to day 7. Mean scores following grading at 24, 48 and 72h were 1.7, 2.0 and 1.3 in erythema and eschar and 1.0, 0.7 and 1.3 for the edema scoring criteria. All effects reversed by day 14. Under the conditions of the study, the test item is not considered to be a skin irritant.

Eye Irritation:

Key study : In vivo, EU Method B.5, 1995 : The study was performed to EU Method B.5 and/or a method equivalent or similar to OECD TG 405 under GLP to assess the irritancy potential of the test item to the eye following a single application in the New Zealand White rabbit. A volume of 0.1 ml of the test item was placed into the left eye of three animals sequentially using the first as a sentinel. The other eye remained untreated and was used for control purposes. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. A single application of the test item to the non-irrigated eye of three rabbits produced no corneal or Iridial inflammation effects. Slight conjunctival reaction (hyperaemia or redness, score = 1 and/or discharge, score = 1) was noted in treated eyes at 1 hour after treatment. No chemosis effects were noted. All treated eyes appeared normal at the 24 hours observation point. Under the conditions of this study, the test item is not considered to be irritating to the eye.

Respiratory Irritation:

No study available

Justification for classification or non-classification

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for dermal irritation.

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for eye irritation.

 

For skin irritation, further in vitro skin corrosion testing does not need to be conducted based on the available information allowing a definitive conclusion on the classification of the substance. The substance does not demonstrate significant skin irritation potential necessary for classification and labelling within an available skin irritation in vivo assay (eq. or similar to OECD TG 404). The substance can cause pronounced transient mild irritation but which is not sufficient for classification and labelling.

 

For eye irritation, the weight of evidence indicates that the substance has the potential to cause transient irritating effects to the eye but which is not sufficient for classification based on the mean scoring and evaluation of the results in three organisms demonstrating that the EU criteria had not been met. Effects in vivo on corneal opacity and iritis are low to non-existent and conjunctival effects are low moderate which fully reversed within 24 hours; the overall evidence is indicative of transient mild effects on the eye.

References:

1. Guidance on Application of the CLP Criteria, ECHA, version 5.0, July 2017