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Toxicological information

Eye irritation

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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:
other: The study was conducted in according to OECD TG 405 and in accordance with the principles of GLP.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
U.S. EPA Health Effects Test Guidelines, OPPTS 870.2400 (1998), JMAFF 12-Nouan-8147, November 2000 and Official Journal of the European Communities. Methods for the Determination of Toxicity, Part B.5 (Eye Irritation), Directive 2004/73/EC, 29 April 2004.
Deviations:
no
Principles of method if other than guideline:
not applicable
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Diethylene glycol mono phenyl ether
- Physical state: Liquid
- Analytical purity: No Certificate of Analysis; Gas Chromatogram indicates 99.5% by area DiEPh
- Lot/batch No.: 200602920-11
- Expiration date of the lot/batch: 19-Feb-2009
- Storage condition of test material: Ambient (+18 to +36°C)
- pH : 3.5 – 7.5 (3% aq. Soln)
- Solubility in water/Miscibility : 3% wt.
- Stability : Test substance was expected to be stable for the duration of testing.

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Toxicology, Department of Safety Assessment, Advinus Therapeutics Private Limited, Bangalore 560 058, INDIA
- Age at study initiation: 5-6 months
- Weight at study initiation: 2.20 to 2.39 kg
- Housing: The rabbits were housed individually in rabbit cages (approx. size: L 65 x B 65 X H 45 cm) with Noryl shallow cage body and noryl perforated raised shelf for enrichment and facilities for pelleted food (Stainless steel feed hopper) and drinking water (750 ml markrolon bottle fitted with sipper tube). The litter collection tray (noryl waste tray) were washed daily with water (except on sundays). The water bottle and feed hopper were changed once a week.
- Diet (e.g. ad libitum): The animals were offered rabbit feed (Nutrilab®) manufactured by Tetragon Chemie Pvt.Ltd., KHB Industrial Area, Yelahanka, New
Town, Bangalore - 64, India.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum
purifier, manufactured by Eureka Forbes Ltd., Mumbai- 400001, was provided to animals in Markrolon bottles with stainless steel sipper tubes.
- Acclimation period: One rabbit RB6926 was acclimatized for 14 days while the other two (RB6927 and RB6928) rabbits were acclimatized for 15 days
under laboratory conditions after physical examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22°C,(except on one day the maximum temperature was 23°C)
- Humidity (%): 30-70%
- Air changes (per hr): 12-15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hour light and 12 hour dark cycle

Test system

Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
Prior to instillation, both eyes of each rabbit were examined with the aid of a pen light for irritation and ocular defects. The eyes were then checked for opacity using ophthalmic fluorescein sodium. One drop was instilled into each eye and the eyes were then irrigated with enough physiological saline (0.9% NaCl) to
remove any excess fluorescein (approximately 30 seconds). Using an ultraviolet lamp, the eyes were checked and evaluated for preexisting corneal damage.
One tenth of a milliliter of the test item (undiluted) was placed in the everted lower lid (conjunctival sac) of the left eye of each rabbit. The eye lids were gently held together for about one second, in order to minimize loss of the test item.
Duration of treatment / exposure:
72 hrs
Observation period (in vivo):
A single rabbit was initially exposed to the test item. Since the test item was not corrosive or severely irritating at 24 hours observation, hence, the test was conducted using two additional animals.
Number of animals or in vitro replicates:
Three female rabbits
Details on study design:
Ocular lesions were evaluated with the illumination of a white light source at intervals of approximately 1, 24, 48 and 72 hours post-instillation of the test substance according to the "Scale for Scoring Ocular Lesions".

In addition, all treated eyes were examined using ophthalmic fluorescein sodium at the 24-hour interval. Corneal opacity exists at 24 hours, hence, fluorescein was used as needed, at subsequent scoring intervals, to aid in evaluating the extent of corneal damage.

Irritation persisted in all rabbits at 72 hours, hence, scoring was continued for all rabbits at 4, 7, 10, 14, 17 and 21 days post instillation.

Individual scores were recorded for each rabbit. All scores were calculated as described in the "Scale for Scoring Ocular lesions",and the total possible points (out of 110) were tallied. In addition to the observation of the cornea, iris and conjunctivae, any other lesions that are observed were noted. To aid in the interpretation of data, the average score for all rabbits at each scoring period was calculated.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
1
Max. score:
1
Reversibility:
not reversible
Remarks:
See Attachment 1
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
2
Max. score:
2
Reversibility:
not reversible
Remarks:
See Attachment 1
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24, 48 and 72 hrs
Score:
1.7
Max. score:
2
Reversibility:
not reversible
Remarks:
See Attachment 1
Irritant / corrosive response data:
The conjunctival redness, chemosis and discharge were observed in all the animals till termination. The corneal opacity was observed in all the animals till termination. There were no iris reactions. Mild circumcorneal injection was observed in one rabbit from day 10 onwards and in the other two rabbits from day 14 onwards till termination of observation on day 21.
Other effects:
The body weights of all rabbits increased slightly through the observation period.

Any other information on results incl. tables

Irritation scores calculated

Cornea opacity
Animal Observation time Mean value Mean value (over all animals)
24 hours 48 hours 72 hours
1 1 1 1 1 1.0
2 1 1 1 1
3 1 1 1 1
Iris lesion
Animal Observation time Mean value Mean value (over all animals)
24 hours 48 hours 72 hours
1 0 0 0 0.0 0.0
2 0 0 0 0
3 0 0 0 0.0
Conjunctivae (redness)
Animal  Observation time Mean value  Mean value (over all animals)
24 hours 48 hours 72 hours
1 2 2 2 2 2.0
2 2 2 2 2.0
3 2 2 2 2
Conjunctivae (chemosis)
Animal  Observation time Mean value Mean value (over all animals)
24 hours 48 hours 72 hours
1 1 1 1 1 1.7
2 2 2 2 2
3 2 2 2 2

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material was irritating to the eyes with conjuctival chemosis and redness and cornea opacity scores meeting the criteria for CLP category 2a. However the cornea opacity effects persisted until the end of the observation period of 21 days. This therefore meets the criteria for a 'serious effect' and as such category 1 eye irritant is appropriate.
Executive summary:

The acute eye irritation/corrosion study in New Zealand White rabbits was conducted to evaluate the eye irritation potential of Diethylene glycol mono phenyl ether. One-tenth of a milliliter (0.1 ml) of the undiluted test item was instilled into the conjunctival sac of the left eye of the animal after gently pulling the lower lid away from the eyeball. The eyelids were held together gently for about one second to minimize loss of test item. The right eye remained untreated and served as the reference control. All the rabbits were treated in a similar manner. The ocular lesions were evaluated at 1, 24, 48, 72 hours, 4, 7, 10, 14, 17 and 21 days post instillation according to the scale for scoring ocular lesions. In addition, the treated eyes were examined using ophthalmic fluorescein sodium at 24, 72 hours, at 7 and 14 days post instillation. The total mean scores were 18, 27.34, 27.34, 27.34, 25.34, 22.06, 34.67, 27.33, 27.33 and 18.67 at 1, 24, 48, 72 hours, 4, 7, 10, 14, 17 and 21 days, respectively. Mild circumcorneal injection was observed in one rabbit from day 10 onwards and in the other two rabbits from day 14 onwards till termination of observation (day 21). There were no toxic signs and pre-terminal deaths.

Based on the evidence of irritation and the persistence of effects until the end of the observation period, the test material meets the criteria for Category 1 A according to CLP.