1-phenylethanol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

data is from experimental reports

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

The objective of the study was to assess the irritant and/or corrosive effects of “1-phenylethanol (CAS No. - 98-85-1)” on eye, when exposed by the ocular route in rabbits.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1-phenylethanol

EC Number:

202-707-1

EC Name:

1-phenylethanol

Cas Number:

98-85-1

Molecular formula:

C8H10O

IUPAC Name:

1-phenylethan-1-ol

Test material form:

liquid

Details on test material:

IUPAC name: 1-phenylethan-1-ol
Mol. formula: C8H10O
Molecular Weight: 122.166 g/mol
Substance form: Colourless liquid
Substance type: organic
InChI: 1S/C8H10O/c1-7(9)8-5-3-2-4-6-8/h2-7,9H,1H3
Smiles : c1(ccccc1)C(C)O

Specific details on test material used for the study:

- IUPAC name: 1-phenylethan-1-ol
- Common Name: 1-Phenylethanol
- Mol. formula: C8H10O
- Molecular Weight: 122.166 g/mole
- InChI: 1S/C8H10O/c1-7(9)8-5-3-2-4-6-8/h2-7,9H,1H3
- Smiles: c1(ccccc1)C(C)O
- Substance type: Organic
- Physical state: Clear colourless liquid
- Analytical purity: 99.62%
- Lot/batch No.: 2014022801R-434
- Storage condition of test material: Room temperature (20 - 30°C)
- Other:
Handling and Disposal
Safety precautions: Aprons, masks, caps, gloves and goggles were used to ensure the health and safety of the personnel.
Disposal: The remaining unused test item was disposed as per internal SOPs and the same was documented in the raw data

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Procured from LIVEON BIOLABS PVT. LTD.
- Sex: Female
- Age at study initiation: 4.5 to 5.0 Months (Approximately)
- Weight at study initiation: Minimum: 2.102 kg and Maximum: 2.652 kg
- Health Status: Healthy young adult rabbits were used. Females were nulliparous and non pregnant.
- Housing: The animals were housed individually in stainless steel cages
- Room Sanitation: The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle: All the cages and water bottles were changed minimum twice a week
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences) ad libitum. Batch No.: 200004.
- Water (e.g. ad libitum): Aqua guard filtered tap water was provided ad libitum
- Acclimation period: Rabbits were acclimatised to the test conditions for a period of 7 days (Animal No.-1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.
- Identification: During acclimatization marking was done with non toxic marker pen in the inside of left ear of rabbits and after acclimatization, animals were marked with permanent number in the inner side of right ear of rabbits. Permanent marker and cage card was used for identification. The individual cage card was labelled with at least study no., study type, test system, sex, dose, experiment start date and experiment completion date.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 18.90°C and Maximum: 22.70 °C
- Humidity (%): Minimum: 48.30% and Maximum: 68.30%
- Air changes (per hr):More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: yes, one eye served as control

Amount / concentration applied:

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

VEHICLE: N/A

Duration of treatment / exposure:

24 hr

Observation period (in vivo):

day 14 (for animal no. 2 ) and day 21 (for animal no. 1 and 3)

Duration of post- treatment incubation (in vitro):

No data available

Number of animals or in vitro replicates:

3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eye of rabbit was washed with normal saline
- Time after start of exposure: 24 hrs

SCORING SYSTEM: Grading of irritation lesions was carried out as per Draize Method

TOOL USED TO ASSESS SCORE: ophthalmoscope

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

fully reversible within: 21 days

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

fully reversible within: 21 days

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 21 days

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal: #1 and #3

Time point:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: 21 days

Remarks on result:

positive indication of irritation

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

The following were observed in treated rabbits.
Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) in animal no. 2 and obvious swelling with partial eversion of lids in animal no. 1 and 3.
Observation at 24 hours after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was seen in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in all the animals.
At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 30%, 30% and 20% damage in animal no. 1, 2 and 3 respectively.
Observation at 48 hours after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was observed in animal no. 1 and 3 whereas some blood vessels definitely hyperaemic (injected) was observed in animal no. 2; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all 3 animals.
Observation at 72 hours after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was observed in animal no. 1 and 3 whereas some blood vessels definitely hyperaemic (injected) was observed in animal no. 2; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all 3 animals.
Observation on day 7 after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in animal no. 1 and 3 and no swelling (Normal) in animal no. 2.
Observation on day 14 after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible was observed in animal no.1 and 3 whereas no ulceration or opacity in animal no. 2; Area of Opacity- One quarter (or less) but not zero was observed in animal no. 1 and 3 whereas zero was observed in animal no.2; Iris: Normal in all the animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in animal no.1 and 3 whereas blood vessels normal was observed in animal no. 2; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in animal no. 1 and 3 whereas no swelling (Normal) was observed in animal no. 2. Animal no. 2 recovered on day 14.
Observation on day 21 after instillation of test item revealed: Cornea- No ulceration or opacity was observed in animal no. 1 and 3; Area of Opacity- Zero was observed in animal no. 1 and 3; Iris: Normal in all the animals; Conjunctivae - Blood vessels normal was observed in animal no. 1 and 3; Chemosis: No swelling (Normal) was observed in animal no. 1 and 3.

Other effects:

Clinical Observation
No systemic toxicity was observed in treated rabbits during the experimental period.
Mortality
No mortality was observed during the observation period.
Body weight
Increase in body weights of all the animals weighed on test day 0 (prior to application) and at termination (when compared to day 0)

Any other information on results incl. tables

Table 1 : Individual Animal Eye Irritation Scores

In Treated area Dose: 0.1 ml of test item (as such)                                               Sex: Female

Animal Number	1
Application Side	Right
Eye Reactions	*	Hour(s)				Day
		1	24	48	72	7	14	21
Cornea	0	0	0	0	0	0	1	0
Area of Opacity	0	0	0	0	0	0	1	0
Iris	0	0	0	0	0	0	0	0
Conjunctiva	0	2	2	2	2	1	1	0
Chemosis	0	2	1	1	1	1	1	0
Corneal Damage (%)	30

Animal Number	2
Application Side	Right
Eye Reactions	*	Hour(s)				Day
		1	24	48	72	7	14
Cornea	0	0	0	0	0	0	0
Area of Opacity	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0
Conjunctiva	0	2	2	1	1	1	0
Chemosis	0	1	1	1	1	0	0
Corneal Damage (%)	30

Animal Numbers	3
Application Side	Right
Eye Reactions	*	Hour(s)				Day
		1	24	48	72	7	14	21
Cornea	0	0	0	0	0	0	1	0
Area of Opacity	0	0	0	0	0	0	1	0
Iris	0	0	0	0	0	0	0	0
Conjunctiva	0	2	2	2	2	1	1	0
Chemosis	0	2	1	1	1	1	1	0
Corneal Damage (%)	20

Key:*= Pre-treatment eye examination

Dose: Untreated (Control Eye)                                                                       Sex:Female

Animal Number	1
Application Side	Left
Eye Reactions	*	Hour(s)				Day
		1	24	48	72	7	14	21
Cornea	0	0	0	0	0	0	0	0
Area of Opacity	0	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0	0
Conjunctiva	0	0	0	0	0	0	0	0
Chemosis	0	0	0	0	0	0	0	0
Corneal Damage (%)	0

Animal Number	2
Application Side	Left
Eye Reactions	*	Hour(s)				Day
		1	24	48	72	7	14
Cornea	0	0	0	0	0	0	0
Area of Opacity	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0
Conjunctiva	0	0	0	0	0	0	0
Chemosis	0	0	0	0	0	0	0
Corneal Damage (%)	0

Animal Number	3
Application Side	Left
Eye Reactions	*	Hour(s)				Day
		1	24	48	72	7	14	21
Cornea	0	0	0	0	0	0	0	0
Area of Opacity	0	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0	0
Conjunctiva	0	0	0	0	0	0	0	0
Chemosis	0	0	0	0	0	0	0	0
Corneal Damage (%)	0

Key:*= Pre- treatment eye examination

Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Treated eye)

Animal No.  Eye Reaction	1	2	3
Corneal Opacity	0.00	0.00	0.00
Iris	0.00	0.00	0.00
Conjunctiva	2.00	1.33	2.00
Chemosis	1.00	1.00	1.00

Formula :

Mean Eye Irritiation score = Sum of the Individual Animal Score for eye reaction at 24, 48 and 72 hours

Number of the Observations (3)

Table 2: Individual Animal Clinical Signs

Animal No.	Days (Post application observation)
	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
2	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	./.	./.	./.	./.	./.	./.	./.
3	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1

Key:1 = Normal

Table 3: Individual Animal Body Weight

Animal No.	Animal Body Weight (kg)
	Prior to application	At termination
1	2.652	2.932
2	2.102	2.290
3	2.614	2.780

Key: kg = Kilogram

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 2.00, 1.00; 0.00, 0.00, 1.33, 1.00, and 0.00, 0.00, 2.00, 1.00, respectively.
Under the experimental conditions tested, all the three animals were fully reversible within an observation period of 14 for animal no. 2. and 21 days for animal no. 1 and 3. Hence, under the experimental test conditions, 1-phenylethanol [CAS: 98-85-1] was considered to be an eye irritant. It was further classified under the category "Category 2".

Executive summary:

Acute eye irritation/corrosion study was conducted in rabbits to evaluate the eye irrritant nature of the test chemical 1-phenylethanol. The study was performed as per OECD 405 Guidelines using 3 female New Zealand White rabbits.

Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 ml of test item was instilled in the other (treated) eye of each rabbit. The eye was observed at 1, 24, 48, 72 hour, day 7, day 14 and day 21 for animal no. 1 and 3 whereas at 1, 24, 48, 72 hour, day 7 and day 14 for animal no. 2 after test item instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test, 0.1 ml of test item (as such) was applied into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3); 0.1 ml of test item was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Untreated eye of all the three rabbits were normal throughout the experimental period. Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) in animal no. 2 and obvious swelling with partial eversion of lids in animal no. 1 and 3.

Observation at 24 hours after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was seen in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was seen in all the animals.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 30%, 30% and 20% damage in animal no. 1, 2 and 3 respectively.

Observation at 48 hours after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was observed in animal no. 1 and 3 whereas some blood vessels definitely hyperaemic (injected) was observed in animal no. 2; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all 3 animals.

Observation at 72 hours after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was observed in animal no. 1 and 3 whereas some blood vessels definitely hyperaemic (injected) was observed in animal no. 2; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all 3 animals.

Observation on day 7 after instillation of test item revealed: Cornea- No ulceration or opacity was seen in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in animal no. 1 and 3 and no swelling (Normal) in animal no. 2.

Observation on day 14 after instillation of test item revealed: Cornea- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible was observed in animal no.1 and 3 whereas no ulceration or opacity in animal no. 2; Area of Opacity- One quarter (or less) but not zero was observed in animal no. 1 and 3 whereas zero was observed in animal no.2; Iris: Normal in all the animals; Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in animal no.1 and 3 whereas blood vessels normal was observed in animal no. 2; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in animal no. 1 and 3 whereas no swelling (Normal) was observed in animal no. 2. Animal no. 2 recovered on day 14.

Observation on day 21 after instillation of test item revealed: Cornea- No ulceration or opacity was observed in animal no. 1 and 3; Area of Opacity- Zero was observed in animal no. 1 and 3; Iris: Normal in all the animals; Conjunctivae - Blood vessels normal was observed in animal no. 1 and 3; Chemosis: No swelling (Normal) was observed in animal no. 1 and 3.

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 2.00, 1.00; 0.00, 0.00, 1.33, 1.00, and 0.00, 0.00, 2.00, 1.00, respectively.

Under the experimental conditions tested, all the three animals were fully reversible within an observation period of 14 for animal no. 2. and 21 days for animal no. 1 and 3. Hence, under the experimental test conditions, 1-phenylethanol [CAS: 98-85-1] was considered to be an eye irritant. It was further classified under the category "Category 2".