(2E)-2-methyl-3-(4-methylphenyl)prop-2-en-1-ol

Endpoint summary

Description of key information

Skin irritation, in vivo: non-irritating, OECD TG 404, 2012

Eye irritation, in vivo: eye irritating, OECD TG 405, 2012

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Data waiving:

other justification

Justification for data waiving:

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

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
In accordance with REACH Regulation (EC) No. 1907/2006 Annex VII, column 2 section 8.1.1 (as amended by Commission Regulation (EU) 2016/863) the in vitro skin corrosion (OECD TG 431) study does not need to be conducted based on the available information allowing a definitive conclusion on the classification of the substance. An available in vivo (OECD TG 404) skin irritation study is available and data in other endpoints (such as skin sensitisation and/or acute dermal toxicity) indicates that the substance is not skin corrosive and a definitive conclusion on the classification can be made. Furthermore, in accordance with section 1.2 of REACH Regulation (EC) No. 1907/2006 Annex XI the weight of evidence indicates that the substance is not skin corrosive and therefore in vitro testing may be omitted. According to ECHA Guidance on Information Requirements and Chemical Safety Assessment (Chapter R.7a: Endpoint Specific Guidance, R.7.2, July 2017) the study does not need to be conducted. Information as to the availability of the in vivo study is provided in 'attached justification'.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28-09-2011 to 18-10-2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Justification for type of information:

Information as to the availability of the in vivo study is provided in 'attached justification'.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

Hsdlf: NZW

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Recognised Supplier
- Age at study initiation: 23 to 27 weeks
- Weight at study initiation: 3608 to 3802 g
- Housing: Individually housed in cages with perforated floors, with shelters and cage enrichment
- Diet (ad libitum): Global Diet 2030 (Recognised Supplier); provided ad libitum (approximately 100 g per day)
- Water (ad libitum): mains drinking water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (actual: 19.1 to 22.0°C)
- Humidity (%): 40 to 70 (actual: 43 – 87%)
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 h light / 12 h dark

IN-LIFE DATES: From: 28-09-2011 To: 18-10-2011

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL. The test item was in a fluid state at the time of weighing, it was not heated and the test item was applied undiluted.
- Concentration (if solution): Not applicable.

VEHICLE
- Amount applied: Not applicable. The test item was in a fluid state at the time of weighing, it was not heated and the test item was applied undiluted.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours (initial observation); additional observations are made daily up to Days 7 and 14 to assess the reversibility of skin reactions (as appropriate).

Number of animals:

3 males

Details on study design:

TEST SITE
- Area of exposure: Dorsal (2 cm x 3 cm metalline patch secured with adhesive tape)
- % coverage: Not reported
- Type of wrap if used: metalline patch secured with adhesive tape and elastic bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes
- Time after start of exposure: Four hours after the application, the dressing was removed and the skin cleaned of residual test item using tap water.

OBSERVATION TIME POINTS
(indicate if minutes, hours or days): 1 hour, 4 hours, 24 hours, 48 hours and 72 hours. Additional observations daily up to 7 or 14 days, as appropriate

SCORING SYSTEM: Draize Scale:
- Method of calculation:
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) *.......................................................................................... 4
*. Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the
maximum grade for erythema (= 4) is given.
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 millimeter) ........................................................... 3
Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure) 4

Irritation parameter:

erythema score

Basis:

mean

Remarks:

n = 3

Time point:

other: 24, 48 , 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

mean

Remarks:

n = 3

Time point:

other: 24, 48, 72 hours

Score:

0.2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Very slight erythema and very slight oedema in the treated skin areas (maximum score = 1 for erythema and edema) in n = 3 males at all observation periods. All effects had resolved in 1 out of 3 in 24 hours. All effects had resolved in 3 out of 3 at 7 days.

Other effects:

All males showed expected gain in body weight during the study.

Table 1. Individual Scores and Mean Scores following 4-hour exposure

Skin Reaction	Reading (hours)	1# Male	#2 Male	3# Male
Erythema/Escar Formation	24	1	1	1
	48	1	1	1
	72	1	1	1
	Total	3.0	3.0	3.0
	Mean	1.0 #	1.0 #	1.0 #
Oedema Formation	24	0	1	1
	48	0	0	0
	72	0	0	0
	Total	0.0	1.0	1.0
	Mean	0.0	0.3 #	0.3 #

 # All effects reversed by day 7

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 to the skin.

Executive summary:

The study was performed to OECD TG 404, EU Method B.4, US EPA OPPTS 870.2500 and Japan (JMAFF) 12 Nousan, Notification No 8147 guidelines to assess the primary skin irritancy potential of the test substance in accordance with GLP in New Zealand White rabbits. The test item was applied sequentially. Three rabbits were exposed for 4-hour by semi-occluded application of the test item to the intact clipped skin with 0.5 mL test item introduced under a 2 cm x 3 cm semi-occlusive patch. The patch was secured in position with adhesive tape and elasticated bandage. After exposure to the test item the patches were removed and individual dose sites were scored at approximately 1, 24, 48, and 72 hours. Very slight erythema (score = 1) and very slight oedema (score = 1) was observed in the treated skin. At 72 hours two males showed erythema located at the edges of the application area. All effects had resolved within 7 days. Odema had resolved within 48 hours. Mean scores for following grading at 24, 48 and 72h were slight in erythema and eschar (score = 1) and zero in oedema (score = 0) scoring criteria. 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

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

other justification

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

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
In accordance with REACH Regulation (EC) No. 1907/2006 Annex VII, column 2 section 8.1.2 (as amended by Commission Regulation (EU) 2016/863) the serious eye damage / eye irritation (OECD TG 437 or OECD TG 438) study does not need to be conducted based on the available information allowing a definitive conclusion on the classification of the substance. An available in vivo (OECD TG 405) eye irritation study is available that indicates that the substance is eye irritating and a definitive conclusion on the classification can be made. According to ECHA Guidance on Information Requirements and Chemical Safety Assessment (Chapter R.7a: Endpoint Specific Guidance, R.7.2, July 2017) the study does not need to be conducted. Information as to the availability of the in vivo study is provided in 'attached justification'.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04-10-2011 to 19-10-2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Justification for type of information:

Information as to the availability of the in vivo study is provided in 'attached justification'.

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Recognised Supplier
- Age at study initiation: 25 to 28 weeks
- Weight at study initiation: 3596 to 3791 g
- Housing: Individually housed in cages with perforated floors, with shelters and cage enrichment
- Diet (ad libitum): Global Diet 2030 (Recognised Supplier); provided ad libitum (approximately 100 g per day)
- Water (ad libitum): mains drinking water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (actual: 19.1 to 22.0°C)
- Humidity (%): 40 to 70 (actual: 43 – 86%)
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 h light / 12 h dark

IN-LIFE DATES: From: 04-10-2011 To: 19-10-2011

Vehicle:

unchanged (no vehicle)

Controls:

other: one eye remained untreated

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL. The test item was in a fluid state at the time of weighing, it was not heated and the test item was applied undiluted.
- Concentration (if solution): undiluted

VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable. The test item was in a fluid state at the time of weighing, it was not heated and the test item was applied undiluted.

Duration of treatment / exposure:

A volume of 0.1 mL of the test item, was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The other eye remained untreated and was used for control purposes. Irrigation of the eye with distilled water or saline, after 1 hour was deemed not necessary or was not reported during the study.

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 (male). Testing was conducted sequentially following testing with a sentinel.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none

SCORING SYSTEM:
The irritation was assessed according to Draize (1977) numerical scoring system. At each observation period, the highest scores given were recorded. Any other ocular effects were also noted.

TOOL USED TO ASSESS SCORE:
- Immediately after the 24-hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. This procedure was repeated to assess recovery.
- Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

n = 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Remarks:

n = 3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

n = 3

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

n = 3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

No corneal effects were noted, all scores were zero.
No iridial effects were noted, all scores were zero.
Moderate conjunctival irritation (score = 2) was noted in all treated eyes 1 h through 72 h after treatment. Chemosis was moderate (score = 2) to minimal (score = 1) at the 24 h through 72 h observation. In all treated eyes and all effects had reversed (score = 0) at the 72 h observation.

Other effects:

All animals showed expected gain in bodyweight during the study.

Table 1.0: Individual and Mean Scores for Cornea, Iris and Conjunctivae

Number and Sex	Time After Treatment	Corneal Opacity		Iridial Inflammation	Conjunctival Redness	Conjunctival Chemosis
1# Male	24 Hours	0		0	2	1
	48 Hours	0	0		2	1
	72 Hours	0	0		2	0
Total		0		0	6	2
Mean		0.0		0.0	2.0 #	0.7 #
#2 Male	24 Hours	0		0	2	1
	48 Hours	0	0		2	1
	72 Hours	0	0		2	1
Total		0		0	6	3
Mean		0.0		0.0	2.0 #	1.0 #
#3 Male	24 Hours	0		0	2	1
	48 Hours	0	0		2	1
	72 Hours	0	0		2	1
Total		0		0	6	1
Mean		0.0	0.0		2.0 #	1.0 #

# All effects reversed by day 7

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the test item is considered irritating to the eye.

Executive summary:

The study was performed to OECD TG 405, EU Method B.5, US EPA OPPTS 870.2400 and Japan (JMAFF) 12 Nousan, Notification No 8147 guidelines 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 conjunctival sac of one eye of three rabbits. The other eye remained untreated and was used for control purposes. The test was conducted in a stepwise manner conducted singularly and then on a further two rabbits in accordance with the guideline. 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 opacity, no iridial inflammation and moderate conjunctival irritation (redness, score = 2) and moderate to slight chemosis (score = 2 or 1) at 24 hours through 72 hours. All treated eyes appeared normal at the 7 day observation (score = 0). Under the conditions of this study, the test item is considered to be irritating to the eye.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin Irritation:

Key data: In vivo, OECD TG 404, 2012: The study was performed to OECD TG 404, EU Method B.4, US EPA OPPTS 870.2500 and Japan (JMAFF) 12 Nousan, Notification No 8147 guidelines to assess the primary skin irritancy potential of the test substance in accordance with GLP in New Zealand White rabbits. The test item was applied sequentially. Three rabbits were exposed for 4-hour by semi-occluded application of the test item to the intact clipped skin with 0.5 mL test item introduced under a 2 cm x 3 cm semi-occlusive patch. The patch was secured in position with adhesive tape and elasticated bandage. After exposure to the test item the patches were removed and individual dose sites were scored at approximately 1, 24, 48, and 72 hours. Very slight erythema (score = 1) and very slight oedema (score = 1) was observed in the treated skin. At 72 hours two males showed erythema located at the edges of the application area. All effects had resolved within 7 days. Odema had resolved within 48 hours. Mean scores for following grading at 24, 48 and 72h were slight in erythema and eschar (score = 1) and zero in oedema (score = 0) scoring criteria. Under the conditions of the study, the test item is not considered to be a skin irritant.

Eye Irritation:

Key data: In vivo, OECD TG 405, 2012: The study was performed to OECD TG 405, EU Method B.5, US EPA OPPTS 870.2400 and Japan (JMAFF) 12 Nousan, Notification No 8147 guidelines 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 conjunctival sac of one eye of three rabbits. The other eye remained untreated and was used for control purposes. The test was conducted in a stepwise manner conducted singularly and then on a further two rabbits in accordance with the guideline. 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 opacity, no iridial inflammation and moderate conjunctival irritation (redness, score = 2) and moderate to slight chemosis (score = 2 or 1) at 24 hours through 72 hours. All treated eyes appeared normal at the 7 day observation (score = 0). Under the conditions of this study, the test item is considered to be irritating to the eye.

 

Respiratory Irritation:

Key data: INHALATION: OECD TG 403, 2013 – The study was performed according to OECD TG 403, EU Method B.2, US EPA OPPTS 870.1300 and Japanese JMAFF guidelines in accordance with GLP to assess the acute inhalation toxicity of the test item. A single group of ten Wistar: Crl:WI(Han) strain rats (five males and five females) were exposed to an aerosol atmosphere of the test item. The groups were exposed for four hours using a nose only exposure system, followed by a fifteen day observation period. The mean achieved atmosphere concentrations were as follows: 5.1 ± 0.1 mg/L based on a nominal concentration of 6.9 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size): > 3.3 μm and < 3.4 μm with geometric Standard Deviation > 1.9 and < 2.0. There was no male and one female mortalities in the 5.1 mg/L mean achieved atmosphere concentration. The single female mortality occurred within 4 hours post-exposure. No clinical signs were noted during exposure. After exposure, the clinical signs observed among the animals were lethargy, tremors, flat posture, hunched posture, uncoordinated movements, slow breathing, laboured respiration, shallow respiration, rales, gasping, piloerection, chromodacryorrhoea and/or ptosis. The surviving animals had recovered from the signs between Days 5 and 6, except for the rales in one male that were also present on Day 8. Body weight loss was noted in all animals during the first week post-exposure. All animals regained weight during the second week. Macroscopic post mortem examination of the female found dead revealed light red discoloration of the lungs. Cloudy eyes were seen in two surviving males. No other abnormalities were found at macroscopic examination. Under the conditions of this study, the inhalation 4h-LC50 (male/female) was considered to be > 5.1 mg/L within the Wistar: Crl:WI(Han) rat. Under the conditions of this study, there were no indications of respiratory irritation.

 

References:

1. OECD TG 403 (2009)

2. OECD 39 (2009)

Justification for classification or non-classification

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

The substance meets classification criteria under Regulation (EC) No 1272/2008 for eye irritation: category 2: H319

 

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 (OECD TG 404).

 

For eye irritation, the weight of evidence indicates that the substance has the potential to cause transient irritating effects to the eye which are sufficient for classification based on the mean scoring and evaluation of the results in three organisms demonstrating that the EU criteria had been met. Effects in vivo on corneal opacity and iritis are low to non-existent and conjunctival effects are low to moderate which fully reversed within 7 days; the overall evidence is indicative of moderate but transient and reversible effects on the eye.

References:

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