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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:
From 09 September 2013 (Start of in-life phase) to 19 December 2013 (GLP compliance statement)
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
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
deviations from the maximum level of daily mean relative humidity : see additional data in results' section
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
other: Food and Agricultural Materials Inspection Centre (FAMIC), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.
Deviations:
not specified
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Trilithium hexafluoroaluminate
EC Number:
237-509-4
EC Name:
Trilithium hexafluoroaluminate
Cas Number:
13821-20-0
Molecular formula:
AlF6.3Li
IUPAC Name:
trilithium(1+) hexafluoroalumanetriuide
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material : Lithium cryolite
- Molecular formula : Li3AlF6
- Molecular weight : 162
- Physical state: White powder
- Storage condition of test material: At room temperature in the dark

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: between 12 and 24 weeks old
- Weight at study initiation: body weights were at least 1.5 kg
- 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 : 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 : Free access to tap water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions

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

IN-LIFE DATES: From: 09 September 2013 To: 03 December 2013.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
The powdery test substance was instilled undiluted as delivered by the sponsor. No correction was made for the purity/composition of the test substance, since the guidelines require a fixed amount to be instilled.
Animals were treated by instillation of, on average, 67.3 mg (range 67.0 – 67.5 mg) of the test substance (a volume of approximately 0.1 mL) in the conjunctival sac of one of the eyes.

VEHICLE
not applicable
Duration of treatment / exposure:
Single instillation.
Observation period (in vivo):
21 days.
Number of animals or in vitro replicates:
3 animals.
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 10 weeks later, after considering the degree of eye irritation observed in the first animal.
Details on study design:
APPLICATION OF THE TEST SUBSTANCE:
Animals were treated by instillation of the test substance in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

REMOVAL OF TEST SUBSTANCE
Immediately after the 1 hour observation, the treated eye was rinsed with approx 50 mL tepid tap water, using a velocity of flow which did not affect the eye, to remove any visible residual test substance. For reference control the other eye was also rinsed.

SCORING SYSTEM:
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 and/or 14 days after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

- 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:
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.
Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.
In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.3
Max. score:
2
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: The highest individual score was 1.
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 24 hours
Remarks on result:
other: The highest individual score was 1.
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 24 hours
Remarks on result:
other: The highest individual score was 1.
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.3
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: The highest individual score was 3.
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.7
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: The highest individual score was 2.
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.7
Max. score:
3
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: The highest individual score was 2.
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: The highest individual score was 3.
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: The highest individual score was 2.
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: The highest individual score was 3.
Irritant / corrosive response data:
Irritation
Instillation of approximately 67 mg of Lithium cryolite (a volume of approximately 0.1 mL) into one eye of each of three rabbits resulted in effects on the iris and conjunctivae.
Iridial irritation grade 1 was observed and resolved within resolved within 24 hours for two animals and within 48 hours in the remaining animal.
The irritation of the conjunctivae consisted of redness, chemosis and discharge. The irritation had completely resolved within 7 days in one animal and within 14 days in the other animals.
No corneal opacity was observed, and treatment of the eye with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage.

Corrosion
There was no evidence of ocular corrosion.
Other effects:
Coloration / Remnants:
Remnants of the test substance were present in the eye on Day 1. No staining of (peri) ocular tissues by the test substance was observed.

Toxicity / Mortality:
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Table 1: Individual eye irritation scores

 

Cornea

Iris

Conjunctivae

Animal

Time after dosing

Opacity

(0-4)

 Area

(0-4)

 Fluor area (%)2

(0-2)

Redness

(0-3)

 Chemosis

(0-4)

 Discharge

(0-3)

Comments

5271

1 hour

0

0

 

1

1

3

3

b

 

24 hours

0

0

0

1

3

3

2

-

 

48 hours

0

0

-

0

2

2

1

-

 

72 hours

0

0

-

0

2

1

0

-

 

7 days

0

0

-

0

1

0

0

-

 

14 days

0

0

-

0

0

0

0

-

 

 

 

 

 

 

 

 

 

 

569

1 hour

0

0

 

1

2

2

2

b

 

24 hours

0

0

0

0

2

2

2

-

 

48 hours

0

0

-

0

2

1

1

-

 

72 hours

0

0

-

0

1

0

0

-

 

7 days

0

0

-

0

1

0

0

-

 

14 days

0

0

-

0

0

0

0

-

 

 

 

 

 

 

 

 

 

 

579

1 hour

0

0

 

1

2

3

3

b

 

24 hours

0

0

0

0

2

2

1

-

 

48 hours

0

0

-

0

2

1

1

-

 

72 hours

0

0

-

0

1

1

1

-

 

7 days

0

0

-

0

0

0

0

-

1 Sentinel

2 Green staining after fluorescein treatment (percentage of total corneal area)

Comments:

b Remnants of the test substance in the eye.

Protocol deviations:

- Deviations from the maximum level of daily mean relative humidity occurred.

Evaluation: Laboratory historical data do not indicate an effect of the deviations. The study integrity was not adversely affected by the deviation.

Standard operating procedures deviations:

Any deviations from standard operating procedures were evaluated and filed in the study file. There were no deviations from standard operating procedures that affected the integrity of the study.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Lithium cryolite does not have to be classified for eye irritation according to the CLP and the UN GHS.
Executive summary:

The acute eye irritation/corrosion of Lithium cryolite in the rabbit was investigated in a study performed according to the OECD Testing Guideline No.405 and under GLP.

Single samples of approximately 67 mg of Lithium cryolite (a volume of approximately 0.1 mL) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours and 7 and/or 14 days after instillation.

Instillation of the test substance resulted in effects on the iris and conjunctivae.

Iridial irritation grade 1 was observed and resolved within 24 hours for two animals and within 48 hours in the remaining animal. The irritation of the conjunctivae consisted of redness, chemosis and discharge. The irritation had completely resolved within 7 days in one animal and within 14 days in the other animals

Remnants of the test substance were present in the eye on Day 1.

For each of the three animals the average scores for three consecutive days (24, 48 and 72h) were calculated separately for the cornea, iris and conjonctiva . The mean scores of the three animals were below the values in CLP for corneal opacity and iritis. For conjonctival redness and oedema, only one animal showed mean values above the values in CLP for classification category 2.

Based on these results Lithium cryolite does not have to be classified for eye irritation according to the CLP and the UN GHS.