Anisole

Endpoint summary

Description of key information

- Skin irritation: slightly irritating to skin but not classified (Key study, reliability 2, OECD no 404)
- Eye irritation: not irritating to eyes (key study, reliability 2, OECD no 405)
- Respiratory irritation: no data

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:

From 31/05/1994 to 20/07/1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study. The purity of the test substance is not indicated.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France.
- Age at study initiation: no data
- Weight at study initiation: 2.7 +/- 0.1 kg
- Housing: individually in polystyrene cages (35 x 55 x 32 cm or 48.2 x 58 x 36.5 cm)
- Diet (e.g. ad libitum): 112 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 3°C
- Humidity (%): 30 to 70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12hrs dark/ 12 hrs light

IN-LIFE DATES: From: To: no data

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Remarks:

the test item is used as supplied by the sponsor (undiluted)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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

VEHICLE: not applicable

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48 and 72 hours after removal of the dressing and then daily until day 15.

Number of animals:

3 males

Details on study design:

TEST SITE
- Area of exposure: right flank (6 cm2)
- % coverage: no data
- Type of wrap if used: the test item was applied to a 6 cm2 dry hydrophilic gauze patch and this was then applied to the right flank of the animals.
The test substance and the gauze patch were held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none. Any residual test substance was observed.
- Time after start of exposure: not applicable

SCORING SYSTEM: OECD recommended scoring system.

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

1.9

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

other: Dryness of the skin

Basis:

animal: all tested animals

Reversibility:

not reversible

Remarks on result:

other: Between days 5 and 15, a dryness of the skin was observed at the treatment site of the three animals

Irritant / corrosive response data:

- Moderate reversible cutaneous reactions were observed after removal of the dressing applied during 4 hours. They consisted of erythema (scores of 1 to 3) and oedema (score of 2). The mean score over 24, 48 and 72 hours for individual animals was 1.7, 1.7 and 2.3 for erythema and 1.3, 1.3 and 1.3 for oedema (see Table 7.3.1/1).
- No ulceration or necrosis was noted.

Other effects:

- A dryness of the skin was observed at the treatment site of one animal on day 4 and daily until day 15 for the three animals.
- Any change in the animal's behaviour was noted.

Table 73.1/1: Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Erythema	Edema
	Max. score: 4	Max. score: 2
60 min	2/1/2	0/0/2
24 h	2/1/2	2/2/2
48 h	2/2/3	2/2/2
72 h	1/2/2	0/0/0
Average 24h, 48h, 72h	1.7/1.7/2.3	1.3/1.3/1.3
Reversibility*)	c	c
Average time (unit) for reversion	7 days	72 hours

 

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

 

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

Under the test conditions of this study, Anisole was slightly irritating to the rabbit skin. The mean score over 24, 48 and 72 hours for individual animals was 1.7, 1.7 and 2.3 for erythema and 1.3, 1.3 and 1.3 for oedema. These cutaneous reactions were fully reversible within 7 days. Based on these results, no classification is required according to the EU legislation (CLP regulation (1272/2008) and Directive 67/548/EEC). According to UN GHS criteria the substance is classified skin. irrit. Cat. 3, H316.

Executive summary:

In a primary dermal irritation study performed according to the OECD guideline No. 404 and in compliance with the GLP, three male

New Zealand rabbits were dermally exposed to 0.5 mL of undiluted Anisole for 4 hours under a semi-occlusive dressing. Cutaneous reactions were observed approximately 1 hour, 24 and 72 hours after removal of the dressing and then daily until day 15. The mean values of the scores for erythema and oedema were calculated for each animal.

Moderate reversible cutaneous reactions were observed after removal of the dressing applied during 4 hours. They consisted of erythema (scores of 1 to 3) and oedema (scores of 2). The mean score over 24, 48 and 72 hours for individual animals was 1.7, 1.7 and 2.3 for erythema and 1.3, 1.3 and 1.3 for oedema. By day 7, no erythema nor oedema were observed.

Between days 5 and 15, a dryness of the skin was observed at the treatment site of the three animals. No ulceration or necrosis was noted.

Under the test conditions of this study, Anisole is considered as slightly irritating to skin. However, according to the classification criteria laid down in Regulation (EC) 1272/2008 (CLP) and in Council Directive 67/548/EEC (and subsequent adaptations), the test item should not be classified as irritating to the skin.

This skin irritation study is classified as acceptable. It satisfies the guideline requirement for a skin irritation in the rabbit.

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:

From 31/05/1994 to 08/07/1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study. The purity of the test substance is not indicated.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France.
- Age at study initiation: no data
- Weight at study initiation: 2.9 +/- 0.2 kg
- Housing: individually
- Diet (e.g. ad libitum): 112 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 3°C
- Humidity (%): 30 to 70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12hrs dark/ 12 hrs light

IN-LIFE DATES: From: To: no data

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): not applicable

VEHICLE: none

Duration of treatment / exposure:

Not rinsed

Observation period (in vivo):

Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after administration of the test substance.

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE: not applicable

SCORING SYSTEM:
Conjunctival lesions and discharge
Chemosis (lids and/or nictitating membranes)
- no swelling . 0
- any swelling above normal (includes nictitating membranes) ....................................................................................1
- obvious swelling with partial eversion of lids ................................................................................................................2
- swelling with lids about half-closed ................................................................................................................................3
- swelling with lids more than half-closed ........................................................................................................................4
Redness (refers to palpebral and bulbar conjunctivae, cornea and iris)
- blood vessels normal .........................................................................................................................................................0
- a number of blood vessels definitely hyperemic (injected) .........................................................................................1
- diffuse, crimson colour, individual vessels not easily discernible .............................................................................2
- diffuse, beefy red ................................................................................................................................................................3
Discharge
- absence of discharge .........................................................................................................................................................0
- slight discharge (does not include small amounts normally found in inner canthus) ............................................1
- discharge with moistening of lids and hairs adjacent to lids .......................................................................................2
- discharge with moistening of lids and hairs on wide area around the eye ................................................................3

Iris lesions
- normal ...................................................................................................................................................................................0
- markedly deepened rugae, congestion, swelling, moderate circum-corneal hyperemia, or injection,
any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive) .....................1
- no reaction to light, haemorrhage, gross destruction (any or all of these)................................................................2

Corneal lesions
Degree of opacity (area most dense taken for reading)
- no ulceration or opacity ......................................................................................................................................................0
- scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible ..1
- easily discernible translucent area, details of iris slightly obscured ...........................................................................2
- nacreous areas, no details of iris visible, size of pupil barely discernible ..................................................................3
- opaque cornea, iris not discernible through the opacity ..............................................................................................4
Area of opacity
- one quarter (or less) but not zero .....................................................................................................................................1
- greater than one quarter but less than a half ..................................................................................................................2
- greater than one half but less than three quarters .........................................................................................................3
- greater than three quarters up to whole area ..................................................................................................................4


TOOL USED TO ASSESS SCORE: fluorescein (batch number 2388)

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.1

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

- After 1 hour, moderate chemosis (scores of 1 or 2), slight redness of the conjunctivae (score of 1) were observed in all 3 animals.
- After 24 hours, a slight chemosis was observed for 1 animal
- After 48 and 72 hours, no ocular reactions were observed.

Other effects:

None

Table 7.3.2/1: Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
60 min	0/0/0	0/0/0	1/1/1	2/2/1
24 h	0/0/0	0/0/0	0/0/0	0/1/0
48 h	0/0/0	0/0/0	0/0/0	0/0/0
72 h	0/0/0	0/0/0	0/0/0	0/0/0
Average 24h, 48h, 72h	0/0/0	0/0/0	0/0/0	0/0.3/0
Reversibility*)	Not applicable	Not applicable	0	c
Average time (unit) for reversion	Not applicable	Not applicable	24 hours	48 hours

 

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions of this study, Anisole is considered as not irritating to eyes and is therefore not classified as eye irritant according to the
EU legislation (CLP regulation (1272/2008) and Directive 67/548/EEC).

Executive summary:

In an eye irritation study performed according to the OECD No. 405 guideline and in compliance with the GLP, 0.1 mL of undiluted Anisole was instilled into the conjunctival sac of the left eye of 3 male New Zealand White rabbits. The eyes were not rinsed after administration of the test substance. Ocular reactions were observed approximately 1, 24, 48 and 72 hours after the administration.

The mean individual score were calculated within 3 scoring times (24, 48 and 72 hours).

Moderate then slight reactions of the conjunctiva were observed after 1 hour and 24 hours, respectively. No iris irritation, no opacity of the cornea were observed (the mean individual scores were 0.0 for all tested animals). The mean individual scores were 0.0/0.3/0.0 for chemosis, however; no ocular reactions were observed 48 and 72 hours after instillation of the test substance.

Under the test conditions of this study, Anisole is considered as non-irritant when administered by ocular route in rabbits. Based on these results, no classification is required according to the EU legislation (CLP regulation (1272/2008) and Directive 67/548/EEC).

This eye irritation study is classified as acceptable. It satisfies the guideline requirement for an eye irritation study in the rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

- Skin irritation: one study was available for this endpoint and considered as the key study (Clouzeau, 1994). This study was performed according to the OECD test guideline No. 404 and in compliance with GLP (Reliability: 2). Three New Zealand rabbits were dermally exposed to 0.5 mL of the test substance in its original form for 4 hours under an semi-occlusive dressing. Animals were observed for 15 days. Irritation was scored using the OECD Recommended Scoring System. The mean score over 24, 48 and 72 hours for individual animals was 1.7, 1.7 and 2.3 for erythema and 1.3, 1.3 and 1.3 for oedema. By day 7, no erythema nor oedema were observed. Between days 5 and 15, a dryness of the skin was observed at the treatment site of the three tested animals. Based on these results, Anisole is considered as slightly irritating to the skin.

- Eye irritation: one study was available for this endpoint and considered as the key study (Clouzeau, 1994). This study was performed according to the OECD test guideline No. 405 and in compliance with GLP (Reliability: 2), 0.1 mL of Anisole was instilled into the conjunctival sac of the left eye of three New Zealand rabbits. The eyes of the animals were not rinsed after instillation. Changes were observed at 1, 24, 48 and 72 hours after dosing and grading according to Draize scale. The mean individual scores (24, 48 and 72 hours) were 0.0 for conjunctival redness, corneal lesions and iridial lesions.The mean individual scores (24, 48 and 72 hours) were

0.0/0.3/0.0 for chemosis which were completely reversible after 2 days. Thus, Anisole is considered as not irritating to the eyes.

- Respiratory irritation: no study available.

Justification for selection of skin irritation / corrosion endpoint:
Only one study was available.

Justification for selection of eye irritation endpoint:
Only one study was available.

Effects on skin irritation/corrosion: slightly irritating

Justification for classification or non-classification

- Skin irritation:

The potential of Anisole to induce dermal irritation was evaluated in rabbits according to OECD 404. The mean score over 24, 48 and 72 hours for individual animals was 1.7/1.7/2.3 for erythema and 1.3/1.3/1.3 for oedema. All these cutaneous effects are reversible within 7 days. Based on these results, Anisole is considered as slightly irritating to the skin but no classification is required according to the CLP Regulation (1272/2008) and the Directive 67/548/EEC criteria.

In this study, the dryness was observed at the treatment site of one animal on day 4 and daily until day 15 for the three animals. In Human, no case of skin dryness was reported following to the exposure to Anisole. In animals, this dryness was sufficiently important to be relevant for classification and leads to the conclusion that a EUH066 (repeated exposure may cause skin dryness or cracking)

according to the CLP Regulation (1272/2008) and a R66 according to the Directive 67/548/EEC criteria is required.

- Eye irritation:

As no signs of eye irritation were observed in an eye irritation study with Anisole conducted according to test guideline No. 405 (Mean scores (24, 48 and 72 hours) were 0.0 for conjunctival redness, corneal lesions and iridial lesions and 0.0/0.3/0.0 for chemosis which were completely reversible after 2 days), Anisole is not classified as irritating to the eyes according to the CLP Regulation (1272/2008) and the Directive 67/548/EEC criteria.

- Respiratory irritation:

No classification is possible due to lack of data.