2-(3,7-dimethylocta-2,6-dienyl)cyclopentan-1-one

Endpoint summary

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:

key study

Study period:

April 18, 1991-May 14, 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

equivalent or similar to guideline

Guideline:

other: Federal Hazardous Substances Act, CFR 16, Section 1500.41

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Buckshire Corp. Perkasie, Pennsylvania 18944, USA
- Housing: Elevated stainless steel with wire mesh flooring, 1 rabbit per cage
- Diet (e.g. ad libitum): Wane 15% Rabbit Ration ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-21 degrees C
- Humidity (%): 40-70%
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark

IN-LIFE DATES: From: April 29, 1991 To: May 14, 1991

Type of coverage:

occlusive

Preparation of test site:

other: one site was left intact, the other was abraded

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

24 hrs

Observation period:

14 days

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: back
- Type of wrap if used: occlusive

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, deionized water

OBSERVATION TIME POINTS
(indicate if minutes, hours or days)
30-60 minutes after end of exposure, 24 and 72 hrs, day 7 and 14

SCORING SYSTEM:
- Method of calculation: Draize

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24/72 hrs

Score:

4.12

Max. score:

8

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 24/72 hrs

Score:

3

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

other: 24/72 hrs

Score:

3

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24/72 hrs

Score:

1.5

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

edema score

Basis:

animal #4

Time point:

other: 24/72 hrs

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

edema score

Basis:

animal #5

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritation parameter:

edema score

Basis:

animal #6

Time point:

other: 24/72 hrs

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Observations were reported at 24 and 72 hrs post exposure.

Irritant / corrosive response data:

All animals showed erythema and edema formation (highest score of 3) at the 24 and 72 hr observations on both intact and abraded skin. All animals also showed edema formation (highest score of 2) at the 24 and 72 hr observations on both intact and abraded skin. All but one animals showed denuded skin and/or exfoliation at the 14 day observation on both intact and abraded skin. No edema was seen on any animal at the day 14 observation.

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

None of the 6 rabbits exhibited eschar formation on day 14 of the observation period. None of the reaction that typify skin corrosive, as noted in the ECHA Guidance, including ulcers, bleeding, bloody scabs, skin discoloration/blanching, alopecia, or hyperkeratosis were reported in any of the animals, and the effects of apritone on the skin were clearly reversible in this study. The test substance is therefore classified as a skin irritant.

Executive summary:

Bioresearch (1991) performed a test for skin corrosion/irritation following the procedures specified in USA Federal Hazardous Substances Act (16CFR 1500.41). Accordingly, Bioresearch (1991) applied apritone to the clipped skin on both sides of the bodies New Zealand White rabbits (n=6). The surface of the skin on one side of each rabbit was abraded through the stratum corneum without disturbing the dermis or causing bleeding, and the skin on the other side was left intact before applying the test article (0.5 ml apritone, neat, to each side). A gauze patch (1 in.²; 6.45 cm²) was then applied to each site of exposed skin, and an occlusive bandage was wrapped around the trunk of each animal and secured with elastic tape. The tape, bandage and gauze were removed after a 24-hour exposure period, and the sites were wiped with deionized water. The sites were scored, using the Draize method approximately 24 hours and again 72 hours after apritone was applied to the skin (i.e., 30 to 60 minutes and again 48 hours after patch removal). 

Bioresearch (1991) presented the primary skin irritation scores of each rabbit. Two of the 6 rabbits scored 3 (moderate to severe erythema ) for erythema/eschar formation at the 24-hour (intact and abraded skin) and 72-hour (abraded skin only) observation points. All other scores were 2 (well defined erythema). None of the rabbits scored 4 (severe erythema to eschar formation preventing grading of erythema) for erythema/eschar formation at either observation point.

One rabbit scored 1 (very slight, barely perceptible) for edema of both intact and abraded skin and another rabbit scored 1 only on the intact skin at the 72-hour observation point. All of the other scores were 2 (slight edema, edges of are well defined by definite raising) at both the 24-hour and 72-hour observation points.

Bioresearch calculated the average erythema/eschar formation scores and edema scores across the 6 rabbits for both intact skin and abraded skin, and then calculated a primary skin irritation index (PII) based on these averages, in accordance with 16CFR 1500.41. Thus, Bioresearch (1991) reported an overall PII of 4.12 for apritone from the results of this study. The PII is 4.08 if only the averages for abraded skin are excluded from the calculation.

The highest average individual scores for were 3 for erythema/eschar formation (in two of the six rabbits) and 2 for edema (in intact skin of 5 of the six rabbits at the 24-hour observation point; in intact skin of 4 of the six rabbits at the 72-hour observation point).

Bioresearch examined and scored the exposed skin again 7-days and 14 days after apritone had been applied, because signs of irritation persisted at the 72-hour observation period. One rabbit scored 1 (very slight, barely perceptible) for edema, and that score was only for the 7-day observation point and was the same for the intact and abraded skin of that single rabbit. One rabbit exhibited remnants of eschar formation on observation day 7, although no score was given for erythema/echar formation in this animal, most probably because the eschar observed was not accompanied by evidence of erythema. All of the other scores were 0 for both erythema/eschar formation and edema at both the 7-day and 14-day observation points.

However, Bioresearch (1991)These authors also noted the following:

• Observation Day 7
• Exfoliation of intact and abraded skin in 4 of 6 animals
• Exfoliation and denudation only on abraded skin in 1 of 6 animals
• Eschar formation in intact and abraded skin in 1 of 6 animals
• Observation Day 14
• Exfoliation of intact and abraded skin in 2 animals
• Exfoliation of intact skin and denudation of abraded skin in 1 animal
• Exfoliation and denudation of both intact and abraded skin in 1 animal
• Denudation of both intact and abraded skin in 1 animal

 

None of the 6 rabbits exhibited eschar formation on day 14 of the observation period.

None of the reaction that typify skin corrosive, as noted in the ECHA Guidance (2017b), including ulcers, bleeding, bloody scabs, skin discoloration/blanching, alopecia, or hyperkeratosis were reported in any of the animals, and the effects of apritone on the skin were clearly reversible in this study.

[1]ECHA (2017b), see Section 3.2.1 Definitions for classification for skin corrosion/irritation, page 271; see also OECD TG 404, Paragraph #23 page 4; see also the U.S. OSHA Hazard Communication Standard (HCS 2012, e.g.,https://www.schc.org/assets/docs/ghs_info_sheets/Skin%20Corrosion%20%20Irritation%20(Final%202018-03).pdf) and the Definitions Chapter 6 of the UN International Labour Organisation (ILO) draft Integrated Proposal (https://www.ilo.org/legacy/english/protection/safework/ghs/ghsfinal/ghsc06.pdf).

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

April 26 - April 29, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

See attachment.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Covance Research Products Inc.
- Age at study initiation: Born December 12, 2009
- Weight at study initiation: 2.9-3.3 kg
- Housing: individually in suspended wire bottom cages
- Diet (e.g. ad libitum): PMI Rabbit Chow daily
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature controlled
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark

IN-LIFE DATES: From: April 26, 2010 To: April 29, 2010

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): 0.5 mL
- Concentration (if solution): 100%

Duration of treatment / exposure:

4 hrs

Observation period:

72 hrs

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 2 x 3 cm
- Type of wrap if used: gauze patch, non-irritating tape, semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with ethanol
- Time after start of exposure: 4 hrs

OBSERVATION TIME POINTS
1, 24, 48, and 72 hrs

SCORING SYSTEM:
- Method of calculation: Erythema and Eschar scale of 0-4, Edema scale of 0-4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hrs

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0.11

Max. score:

8

Reversibility:

fully reversible within: 72 hrs

Irritant / corrosive response data:

One animal showed an erythema and eschar score of 1 at the 48 hr observation. This had fully resolved by the 72 hr observation.

Other effects:

None

Dermal Observations

 

	Animal H3200	Animal H3202	Animal H3203
Erythema and Eschar – 1 hr	0	0	0
Edema – 1 hr	0	0	0
Erythema and Eschar – 24 hrs	0	0	0
Edema - 24 hrs	0	0	0
Erythema and Eschar – 48 hrs	0	0	1
Edema - 48 hrs	0	0	0
Erythema and Escahr – 72 hrs	0	0	0
Edema – 72 hrs	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance is not irritating to skin.

Executive summary:

The skin irritation potential of the substance methyl apritone was determined by exposing three rabbits dermally to 0.5 mL of the substance for 4 hrs. One animal showed mild erythema at the 48 hr observation, which was fully resolved by the 72 hr observation. No other signs of irritation were noted. Methyl apritone is therefore not irritating to skin.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

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

Adequacy of study:

supporting study

Study period:

April 26 - April 29, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

See attachment.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Covance Research Products Inc.
- Age at study initiation: Born December 12, 2009
- Weight at study initiation: 2.9-3.3 kg
- Housing: individually in suspended wire bottom cages
- Diet (e.g. ad libitum): PMI Rabbit Chow daily
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature controlled
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark

IN-LIFE DATES: From: April 26, 2010 To: April 29, 2010

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): 0.5 mL
- Concentration (if solution): 100%

Duration of treatment / exposure:

4 hrs

Observation period:

72 hrs

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 2 x 3 cm
- Type of wrap if used: gauze patch, non-irritating tape, semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with ethanol
- Time after start of exposure: 4 hrs

OBSERVATION TIME POINTS
1, 24, 48, and 72 hrs

SCORING SYSTEM:
- Method of calculation: Erythema and Eschar scale of 0-4, Edema scale of 0-4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hrs

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0.11

Max. score:

8

Reversibility:

fully reversible within: 72 hrs

Irritant / corrosive response data:

One animal showed an erythema and eschar score of 1 at the 48 hr observation. This had fully resolved by the 72 hr observation.

Other effects:

None

Dermal Observations

 

	Animal H3200	Animal H3202	Animal H3203
Erythema and Eschar – 1 hr	0	0	0
Edema – 1 hr	0	0	0
Erythema and Eschar – 24 hrs	0	0	0
Edema - 24 hrs	0	0	0
Erythema and Eschar – 48 hrs	0	0	1
Edema - 48 hrs	0	0	0
Erythema and Escahr – 72 hrs	0	0	0
Edema – 72 hrs	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

Methyl apritone is not irritating to skin. Apritone is therefore not expected to be irritating to skin.

Executive summary:

The skin irritation potential of the substance methyl apritone was determined by exposing three rabitts dermally to 0.5 mL of the substance for 4 hrs. One animal showed mild erythema at the 48 hr observation, which was fully resolved by the 72 hr observation. No other signs of irritation were noted. Methyl apritone is not irritating to skin. Apritone is therefore not expected to be irritating to skin.

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:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Covance Research Products, Inc., Denver Pennsylvania
- Age at study initiation: Approx. 3 months
- Weight at study initiation: 3.0-3.3 kg
- Housing: Suspended cages, 1 per cage
- Diet (e.g. ad libitum): PMI Rabbit Chow daily
- Water (e.g. ad libitum): ad libitum


ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark

IN-LIFE DATES: From: April 27, 2010 To: May 7, 2010

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): as received

Duration of treatment / exposure:

72 hrs

Observation period (in vivo):

72 hrs

Number of animals or in vitro replicates:

3 (1 male/ 2 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not done

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein was used at 24 hr observation

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritant / corrosive response data:

No sign of irritation was noted in the eyes of any of the animals at any of the observations.

Other effects:

- Other observations: No abnormal physical signs were noted in the observation period.

Eye Irritation Scores

Animal		1 hr	24 hr	48 hr	72 hr
H3236	Conjunctiva - Erythema	0	0	0	0
	Conjunctiva – Chemosis	0	0	0	0
	Iris	0	0	0	0
	Cornea	0	0	0	0
H3210	Conjunctiva - Erythema	0	0	0	0
	Conjunctiva – Chemosis	0	0	0	0
	Iris	0	0	0	0
	Cornea	0	0	0	0
H3240	Conjunctiva - Erythema	0	0	0	0
	Conjunctiva – Chemosis	0	0	0	0
	Iris	0	0	0	0
	Cornea	0	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

Methyl apritone did not produce irritation in the eyes of rabbits. The substance is not irritating to the eye. Thus, the GHS criteria for classification are not met.

Executive summary:

The eye irritation potential of methyl apritone was tested in an OPPTS Series 870.2400 experiment. 0.1 mL of test substance was added to the eyes of three rabbits. The untreated eye was used as a negative control. Observations of the animals were performed at 1, 24, 48, and 72 hrs post dosing. No signs of irritation were seen in any animal at any of the observations. The test substance is therefore not irritating to the eye. The GHS criteria for classification is not met.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

See attachment.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Covance Research Products, Inc., Denver Pennsylvania
- Age at study initiation: Approx. 3 months
- Weight at study initiation: 3.0-3.3 kg
- Housing: Suspended cages, 1 per cage
- Diet (e.g. ad libitum): PMI Rabbit Chow daily
- Water (e.g. ad libitum): ad libitum


ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark

IN-LIFE DATES: From: April 27, 2010 To: May 7, 2010

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): as received

Duration of treatment / exposure:

72 hrs

Observation period (in vivo):

72 hrs

Number of animals or in vitro replicates:

3 (1 male/ 2 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not done

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein was used at 24 hr observation

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritant / corrosive response data:

No sign of irritation was noted in the eyes of any of the animals at any of the observations.

Other effects:

- Other observations: No abnormal physical signs were noted in the observation period.

Eye Irritation Scores

Animal		1 hr	24 hr	48 hr	72 hr
H3236	Conjunctiva - Erythema	0	0	0	0
	Conjunctiva – Chemosis	0	0	0	0
	Iris	0	0	0	0
	Cornea	0	0	0	0
H3210	Conjunctiva - Erythema	0	0	0	0
	Conjunctiva – Chemosis	0	0	0	0
	Iris	0	0	0	0
	Cornea	0	0	0	0
H3240	Conjunctiva - Erythema	0	0	0	0
	Conjunctiva – Chemosis	0	0	0	0
	Iris	0	0	0	0
	Cornea	0	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

The source substance methyl apritone did not produce irritation in the eyes of rabbits. The substance is not irritating to the eye. Thus, the GHS criteria for classification are not met. Apritone is therefore not expected to be irritating to the eye.

Executive summary:

The eye irritation potential of the source substance methyl apritone was tested in an OPPTS Series 870.2400 experiment. 0.1 mL of test substance was added to the eyes of three rabbits. The untreated eye was used as a negative control. Observations of the animals were performed at 1, 24, 48, and 72 hrs post dosing. No signs of irritation were seen in any animal at any of the observations. The test substance is therefore not irritating to the eye. The GHS criteria for classification is not met. Apritone is therefore not expected to be irritating to the eye.

Reference 3

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January 28 - August 1, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes (incl. QA statement)

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: abattoir A. Moksel AG, Buchloe, Germany
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Eyes were transported on ice with antibiotics.
- Time interval prior to initiating testing: Eyes were prepared immediately after arrival.
- indication of any existing defects or lesions in ocular tissue samples: Eyes were examined for defects, and any eyes with defects were discarded.
- Indication of any antibiotics used: Penicillin and streptomycin

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): 750 uL
- Concentration (if solution): 100%

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

3.5 hrs

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
Tissue around the eyeball was carefully pulled away. Cornea was excised to 2-3 mm rim of sclera. These were then stored in petri dishes containing HBSS. They were then mounted in corneal holders. The corneal holders were filled with RPMI 1640 medium without phenol red, but with 1% FBS and 2 mM L-glutamine. Posterior chambers were filled first. Corneas were then incubated for 1 hr at 32 degrees C.

QUALITY CHECK OF THE ISOLATED CORNEAS
Yes

NUMBER OF REPLICATES
3

NEGATIVE CONTROL USED
Physiological saline solution (0.9% NaCl)

POSITIVE CONTROL USED
100% ethanol

APPLICATION DOSE AND EXPOSURE TIME
750 uL for 10 minutes

POST-INCUBATION PERIOD: yes, 3.5 hrs

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 3 times
- POST-EXPOSURE INCUBATION: 3.5 hrs, with illuminance measurement after 2 hrs, new medium introduced, and then additional incubation for 90 minutes and the optical density measured.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Illuminance, optical density
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry
- Others (e.g, pertinent visual observations, histopathology): histopathology

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: IVIS =< 3, no category
IVIS >3, =< 55, No prediction can be made
IVIS > 55, cateogry 1

Irritation parameter:

in vitro irritation score

Run / experiment:

7

Value:

2.52

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

8

Value:

1.15

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

9

Value:

1.42

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

Average

Value:

1.77

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system:

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Range of historical values if different from the ones specified in the test guideline:

Histopathology showed at most mild irritation.

Interpretation of results:

GHS criteria not met

Conclusions:

The mean in vitro irritation score was 1.77. By UN GHS guidelines, the substance is not categorized as an eye irritant.

Executive summary:

The eye irritancy potential of apritone was tested in an OECD guideline 437 Bovine Corneal Opacity and Permeability Assay. Three fresh bovine eyes were exposed to 750 uL of the test substance for 10 minutes. The eyes were then washed, and the optical density was determined after 3.5 hrs of incubation. Physiological saline was used as a negative control, and 100% ethanol was used as a positive control. Negative and positive control results were valid. The mean in vitro irritation score was determined to be 1.77. By UN GHS guidelines, the substance is not categorized as an eye irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Justification for classification or non-classification

See attachment.