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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
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Version / remarks:
1998
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF guidelines; including the most recent partial revisions.
Version / remarks:
2011
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: F-12676, PDC lot 20
- Expiration date of the lot/batch: 03 September 2013 (allocated by WIL Research Europe B.V., 1 year after receipt of the test substance)
- Purity test date: 99.96%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature in the dark

FORM AS APPLIED IN THE TEST (if different from that of starting material): Test material was applied unchanged

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: At least 6 weeks old
- Weight at study initiation: 2010 g (average)
- Housing: Animals were individually housed in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32 x 23 cm)
- Diet (e.g. ad libitum): Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) and wooden sticks (Swedish aspen wood, Bioservices, Uden, The Netherlands) were available during the study period
- Water (e.g. ad libitum): Free access to tap water
- Acclimation period: Acclimatization period was at least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40-70%
- Air changes (per hr): Approximately 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12:12 light:dark cycle

IN-LIFE DATES: From: 19 November 2012 To: 03 December 2012

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 64 mg (a volume of approximately 0.1 mL) unchanged
Duration of treatment / exposure:
The lids were held together for about one second to prevent loss of the test substance. The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 7 days later, after considering the degree of eye irritation observed in the first animal
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
3 males
Details on study design:
SCORING SYSTEM:
CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal luster) ............................................ 0
Scattered or diffuse areas of opacity, details of iris clearly visible................................................. 1
Easily discernible translucent area, details of iris slightly obscured............................................... 2
Nacreous area, no details of iris visible, size of pupil barely discernible........................................ 3
Opaque cornea, iris not discernible through the opacity .............................................................. 4
Area of cornea involved:
No ulceration or opacity................................................................................................................ 0
One quarter or less but not zero .................................................................................................. 1
Greater than one quarter, but less than half ................................................................................. 2
Greater than half, but less than three quarters ............................................................................. 3
Greater than three quarters, up to whole area .............................................................................. 4

IRIS
Normal ........................................................................................................................................ 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia,
or injection, any of these or combination thereof, iris still reacting to light
(sluggish reaction is positive) ....................................................................................................... 1
No reaction to light, hemorrhage, gross destruction (any or all of these)....................................... 2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal .................................................................................................................. 0
Some blood vessels definitely hyperaemic (injected).................................................................... 1
Diffuse, crimson color, individual vessels not easily discernible.................................................... 2
Diffuse beefy red.......................................................................................................................... 3

Chemosis (refers to 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

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals) ........... 0
Any amount different from normal and/or lacrimation ................................................................... 1
Discharge with moistening of the lids and hairs just adjacent to lids ............................................. 2
Discharge with moistening of the lids and hairs (considerable area around the eye) ................... 3

TOOL USED TO ASSESS SCORE: Immediately after the 24-hour observation, a solution of 2% fluorescein (Merck, Darmstadt, Germany) in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. This procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area. Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: Fluor area (%)
Basis:
animal #1
Time point:
24 h
Score:
0
Irritation parameter:
other: Fluor area (%)
Basis:
animal #2
Time point:
24 h
Score:
35
Max. score:
35
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
other: Fluor area (%)
Basis:
animal #3
Time point:
24 h
Score:
0
Irritation parameter:
cornea opacity 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:
0.7
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Irritation parameter:
iris score
Basis:
animal: #1, #2, #3
Time point:
24/48/72 h
Score:
0
Irritation parameter:
other: conjunctival redness
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.7
Irritation parameter:
other: conjunctival redness
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Irritation parameter:
other: conjunctival redness
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.3
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.3
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.3
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.7
Irritant / corrosive response data:
Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 1) and epithelial damage (maximum 35% of the corneal area) in one animal, which had resolved within 72 hours. Slight dulling of the normal luster of the cornea was noted in two animals on Day 1. Iridial irritation grade 1 was observed in all animals and resolved within 24 hours. The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 7 days. There was no evidence of ocular corrosion.
Other effects:
No staining of (peri) ocular tissues by the test substance was observed. Test substance remnants were seen in the eye of one animal on Day 1 only. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
 Based on the results of the study, the test article is a mild ocular irritant, and has no obligatory labeling requirements for eye irritation according to GHS.
Executive summary:

The primary eye irritation/corrosion potential of the test article was evaluated in New Zealand White Rabbits. The study was conducted in compliance with OECD GLP (1997). The test method was based on OECD No. 405 (2002), EC No. 440/2008, EPA OPPTS 870.2400 (1998) and JMAFF guidelines (2011). Single samples of approximately 64 mg of test article (a volume of approximately 0.1 mL) were instilled in the conjunctival sac of one of the eyes of each animal. The lids were held together for about one second to prevent loss of the test substance. The remaining untreated eye served as the control. The eyes were observed at 1, 24, 48, and 72 hours and 7 days after instillation. Immediately after the 24-hour observation, a solution of 2% fluorescein in water was instilled in both eyes of each animal to quantitatively determine corneal epithelial damage. Corneal opacity (maximum grade 1) and epithelial damage (maximum 35% of the corneal area) that had resolved within 72 hours was observed in one animal. Slight dulling of the normal luster of the cornea was noted in the other two animals on Day 1. Iridial irritation (grade 1) was observed in all animals and resolved within 24 hours. Conjunctivae redness, chemosis, and discharge (maximum grade 2) was seen in all animals but had resolved within 7 days. Individual animal average corneal opacity scores (24-72 hours) were 0.7, 0.0, and 0.0. Individual animal average iritis scores (24-72 hours) were 0.0 in all animals. Individual animal average conjunctival redness scores (24-72 hours) were 2.0, 1.7, and 1.3. Individual animal average conjunctival chemosis scores (24-72 hours) were 1.3, 0.3, and 0.7. Based on the results of the study, the test article is a mild ocular irritant and has no obligatory labeling requirements for eye irritation according to CLP.