Benzyl cinnamate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04.10.1999 - 08.10.1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Feb. 1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EEC Guideline B.5 "Acute Toxicity (Eye Irritation)"

Version / remarks:

Jan. 1997

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Benzyl cinnamate

EC Number:

203-109-3

EC Name:

Benzyl cinnamate

Cas Number:

103-41-3

Molecular formula:

C16H14O2

IUPAC Name:

benzyl 3-phenylacrylate

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight: 2.6 - 3.0 kg b.w.
- Housing: individually in PPO cages (floor area: 2576 cm2) with performed floor.
- Diet: pelleted complete rabbit diet "Altromin 2123" from Altromin, D-32791 Lage, Lippe; ad libitum.
- Water: free access to domestic drinking water acidified with hydrochloric acid to pH 2.5
- Acclimation period: at least 1 week

ENVIRONMENTAL CONDITIONS
- Temperature: 20°C ± 3°C
- Humidity: 55% ± 15%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): cycle of 12 hours light (06 to 18h) and 12 hours darkness

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

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

Duration of treatment / exposure:

Single exposure without rinsing.

Observation period (in vivo):

1, 24, 48 and 72 hours after treatment

Number of animals or in vitro replicates:

4 females

Details on study design:

TESTING PROCEDURE
- The day before testing both eyes of the animals were examined with a hand held inspection lamp fitted with white and UV-light and magnifying glass with 2 x magnification to ensure there were no defects or irritation. The examination was performed before and after instillation of Fluorescein.

- The left eye was treated. The right eye remained untreated and served as control.
- 0.1 ml of the warmed test item was placed in the left eye of the rabbits by gently pulling the lower lid away from the eyeball to form a cup into which the test article was dropped. The lids were then gently held together for 1 second.

- The eyes were examined and the grade of ocular reaction was recorded one hour and 24 hours later. After the first 24 hour reading Fluorescein was instilled. After rinsing with 20 ml 0.9% sodium chloride solution the eyes were examined again using UV-light to detect possible comeal damage.
- The eyes were also examined 48 and 72 hours after the treatment.

EVALUATION OF THE REACTIONS (see tables below)

ASSESSMENT
A substance or a preparation is considered to be irritant if it causes ocular lesions which occur within 72 hours after exposure and which persist for at least 24 hours.

Ocular lesions are significant if the rnean scores of the eye irritation test have any of the following values:
- cornea opacity ≥ 2 but < 3
- iris lesion ≥ 1 but < 1.5
- redness of conjunctivae ≥ 2.5
- oedema of conjunctivae (chemosis) ≥ 2
or, in the case where the test has being completed using three animals if the lesions, on two or more animals, are equivalent to any of the above values except that for iris lesion the value should be equal or greater than 1 but less than 2 and for redness of the conjunctivae the value should be equal or greater than 2.5.
In both cases all scores at each of the reading times (24, 48 and 72 hours) for an effect should be used in calculating the respective mean values.

A substance or a preparation is considered to be able to cause serious damage to eyes if it causes severe ocular lesions which occur within 72 hours after exposure and which persist for at least 24 hours.

Ocular lesions are severe if the means of the scores of the eye Irritation test have any of the values:
- cornea opacity ≥3
- iris lesion ≥ 1.5
The same shall be the case where the test has been completed using three animals if these lesions, on two or more animals, have any of the values:
- cornea opacity ≥ 3
- iris lesion = 2
In both cases all scores at each of the reading times (24, 48 and 72 hours) for an effect should be used in calculating the respective mean values.

Ocular lesions are also severe when they are still present at the end of the observation time.

Ocular lesions are also severe if the substance or preparation causes irreversible colouration of the eyes.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3, #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Results 2

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3, #4

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: reversibility: not applicable

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2, #3, #4

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3, #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritant / corrosive response data:

One hour after application of the test article animals No. 1565, No. 1571, No. 1572 and No. 1573 showed some conjunctival vessels definitely injected.
After 24 hours some conjunctival vessels were definitely injected in all animals. After 48 and 72 hours all animals were free of any signs of eye irritation.

Any other information on results incl. tables

Scores for ocular lesions:

*individual mean score: Only the scores from the readings after 24, 48 and 72 hours are included in the calculation of the individual mean scores.

Rabbit No/Weight (kg)	Parameters	Individual mean score*
1565/2.9	cornea opacity	0.00
	iris	0.00
	conjunctiva redness	0.33
	conjunctiva chemosis	0.00
1571/2.6	cornea opacity	0.00
	iris	0.00
	conjunctiva redness	0.33
	conjunctiva chemosis	0.00
1572/2.8	cornea opacity	0.00
	iris	0.00
	conjunctiva redness	0.33
	conjunctiva chemosis	0.00
1573/3.0	cornea opacity	0.00
	iris	0.00
	conjunctiva redness	0.33
	conjunctiva chemosis	0.00

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

The test substance shall not be classified as eye irritating.

Executive summary:

In the current study the eye irritant effect of the test item was investigated according to the method recommended in the OECD Guideline 405 and EEC Guideline B.5.

Four female albino rabbits were exposed to 0.1 mL of the undiluted test articie in the left eye, while the other eye remained untreated and served as control. The eyes were examined and the changes were graded according to a numerical scale one hour, 24, 48 and 72 hours after dosing.

Slight signs of irritation were observed on the treated eyes, however, these were reversible and according to the criteria set out in the CLP regulation the test item shall not be classified as eye irritant.