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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
April 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Substance name: Iron dichloride (Ⅱ)(CAS No.7758-94-3)
Lot No.: 23828CB
Acceptance date: February 11, 2004
Appearance: Pale brown powder
Purity: 98%
Storage conditions: Brown bottle, dry place, sealed

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: : Samtaco Bio Korea, 77-1 Seorang-dong, Osan-si, Gyeonggido, Korea

- Age at study initiation: 4 to 5 months old
- Weight at study initiation: 2200 ~2400g
- Housing: Stored individually in cages
- Diet (e.g. ad libitum): The animals were fed rabbit feeds (Purina Korea located in 85-1 Jangdang-dong, Pyeongtaek, Gyeonggido)
- Water (e.g. ad libitum): The water was sterilized with UV.
- Acclimation period: Not stated

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature was set at 20±2℃
- Humidity (%): relative humidity at 50±10 %
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): lighting time at 12 hours (8 a.m. to 8 p.m.) and illumination at 200 to 300 Lux.

IN-LIFE DATES: From: Day 0 To: Day 14

Test system

Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The powder was finely ground and weighed on the day of testing. 0.1 g of the powder of iron dichloride (Ⅱ)(CAS No.7758-94-3) was applied to the conjunctiva of the eye.


According to MSDS, LD 50 was 450 mg/kg in the case of oral administration and 59 mg/kg in the case of intraperitoneal injection. Therefore, as irritations by the test substance were anticipated, an initial test was conducted on one rabbit before the confirmative test was conducted.

VEHICLE
Not applicable no vehicle used
Duration of treatment / exposure:
The test substance was applied once.
Observation period (in vivo):
Temperature and humidity were automatically measured in the animal chamber every hour during the test period.
Clinical signs, poisoning and mortality were checked till 72 hours from the application. Continued eye irritation was observed, so the observation lasted till Day 21.

The presence of irritation on the cornea, iris and conjunctiva was checked with the both-eye real microscope (Leica S6E, Germany) at 1 hour, 24 hours, 48 hours, and 72 hours after the application and was observed up to 21 days.
Number of animals or in vitro replicates:
3 females
Details on study design:
SCORING SYSTEM:
Evaluation of eye irritation and irritation determination (KOTRIC-EIT-001)
The determination of irritation was based on the table 1 ‘Grade of eye lesion’. Eye lesion scores were calculated at each observation time depending on the degree of eye lesion. The scores must not be used as criteria. Each score can be meaningful when it was suggested with all the observation items and descriptions. The eye irritation scores were calculated as with the severity and characteristics of the lesion, reversibility or reversibility loss.

Grade of eye lesion
(1) Cornea
(A) Diffuse area of opacity: degree of opacity (Observe the most opaque area)
◦ No ulcer or opacity
◦ Opacity is scattered or distributed (different from a little spoiled normal transparency), but the iris is clearly observed.
◦ Semi-opaque areas are easily observed, but the fineness of the iris is a bit unclear.
◦ Pearly color, and the iris is not finely observed. The size of the pupil is barely observed.
◦ The cornea is opaque and turbid, so the iris cannot be distinguished.

(2) Iris
◦ Normal
◦ Forms deep wrinkles. Hyperemia, Edema, Single or complicated hyperemia of moderate severity around the cornea, delayed response of the iris to light (Delayed response is positive.)
◦ No response to light, bleeding, broad destruction (some of the symptoms mentioned above or all)

(3) Conjunctiva
A. Redness (eyelid, conjunctiva, cornea, iris)
◦ Normal blood vessel
◦ Clear hyperemia in a few blood vessels
◦ Wide-spread scarlet color, blood vessels not easily distinguished
◦ Wide-spread crimson red

B. Conjunctival edema (eyelid and/or nictitating membrane)
◦ Not swollen
◦ A little more edema than normal (Including nictitating membrane)
◦ Outstanding edema with partial eversion of the eyelid
◦ Edema of eyelid as severe as eyes are half closed
◦ Edema of eyelid as severe as eyes are closed more than half

TOOL USED TO ASSESS SCORE:
The tool used to assess the score was the both-eye real microscope (Leica S6E, Germany)

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal 1
Time point:
other: 24, 48 and 72 hours
Score:
2.33
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal 2
Time point:
other: 24, 48 and 72 hours
Score:
3
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal 3
Time point:
other: 24, 48 and 72 hours
Score:
3.3
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal 1
Time point:
other: 24, 48 and 72 hours
Score:
1.33
Max. score:
2
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal 2
Time point:
other: 24, 48 and 72 hours
Score:
1.66
Max. score:
2
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal 3
Time point:
other: 24, 48 and 72 hours
Score:
1.66
Max. score:
2
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal 1
Time point:
other: 24, 48 and 72 hours
Score:
2.33
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal 2
Time point:
other: 24, 48 and 72 hours
Score:
2.33
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal 3
Time point:
other: 24, 48 and 72 hours
Score:
2.66
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
other: Edema score
Basis:
mean
Remarks:
animal 1
Time point:
other: 24, 48 and 72 hours
Score:
2.66
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
other: Edema score
Basis:
mean
Remarks:
animal 2
Time point:
other: 24, 48 and 72 hours
Score:
3.66
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
other: Edema score
Basis:
mean
Remarks:
animal 3
Time point:
other: 24, 48 and 72 hours
Score:
4
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritant / corrosive response data:
In the initial test, irritation was observed in the cornea, iris and conjunctiva in the applied region at 1 hour. So two more animals were tested in the confirmative test and the applied regions were observed. As a result of observation at 1 hour, and Days 1, 2, 3, 7, 14, and 21, the opacity was observed in the cornea of all the animals at all observation times. At Day 21, one rabbit was observed to be in the first stage recovery and the opacity did not disappear in the rest two rabbits. All the animals did not show the response of iris to light at Days 2-3 and hyperemia was observed in all of them. Severe redness (Grade 1 ~ 3), severe excretion and yellow pigmentation were observed in the conjunctiva of all the subjects. Severe edema (Grade 2 ~4) was observed in the conjunctiva and nictitating membrane of all the animals. The redness and edema began to relieve in Animal No. 1 at Day 7, in Animal No. 2 at Day 4 and Animal No. 3 at Day 14. With the relieved edema, the deformation of upper and lower eyelids was observed.
Other effects:
Clinical signs and mortality:
All the animals showed reduced mobility and two animals were observed to rub the eye with front paws. No mortality was observed. Head tilting was observed in one rabbit in the initial test and one rabbit in the confirmative test.

Body weight change:
All the animals weighed more at Day 21 after the application than at Day 0.

Any other information on results incl. tables

After 0.1 g of the test substance was applied in the conjunctiva to study the eye irritation of iron dichloride(Ⅱ)(CAS No.7758-94-3) in three New Zealand White female rabbits. Mortality, clinical signs and body weight changes were observed for 21 days and the eye irritation was evaluated at 1 hour, and Days 1, 2, 3, 7, 14 and 21. During the test period, all the animals showed reduced mobility and two rabbits were observed to rub their eyes with front paws. This was the reaction of rabbits sensitive to the eye irritation because the substance irritates the mucosa of the eye. No mortality was observed during the test period. Head tilting was observed in the rabbit in the initial test and one rabbit in the confirmative test. It seemed to be caused by the substance that melted in tear and flowed into the nasal cavity through the lacrimal gland. The histopathological examination found that lymphocyte nodule was formed in the submucosa of the paranasal sinus and the test substance solution affected the nasal cavity tissue through the lacrimal gland. All the animals were observed to weigh more at Day 21 than Day 0, which indicated the test substance did affect topical irritation but not the whole body.

As a result of observing the applied region at 1 hour, and Days 1, 2, 3, 7, 14, and 21, an opaque area was observed throughout the cornea of all the animals at each time. One rabbit showed stage one recovery at Day 21 and the opacity did not disappear in the rest two rabbits. All the animals showed no response of the iris to light at Days 2 ~ 3 and hyperaemia was observed. Severe redness (Grade 1-3), severe excretion and yellow pigmentation were observed in the conjunctiva of all the subjects. Severe edema (Grade 2-4) was found in the conjunctiva and nictitating membrane of all the animals. The redness and edema began to relieve in Animal No. 1 at Day 7, Animal No. 2 at Day 4 and Animal No. 3 at Day 14. However, with the relieved edema, the deformation of upper and lower eyelids was observed, which indicated that the test substance caused severe purulent and inflammatory lesion because hemosiderosis and severe lymphocyte infiltration were found in the dermis in the deformed eyelids. High scores appeared even in the determination based on the Grading of eye lesion.

As a result of the histopathological examination on eyeball, conjunctiva and lesion of eyelid, granulomatous inflammation was observed in the conjunctiva, so necrosis, deformed collagen fiber and mineralization were found in the giant cell and the central part. As purulent keratitis made of neutrophils was observed in the stroma of the cornea, the test substance seemed to cause deformation in the cornea and conjunctiva. Given the fact that the iris had no histopathological lesion, the symptom in the iris was thought to disappear within 21 days. Corneal perforation, severe corneal ulcer including staphyloma, hyphema, conjunctival ulcer, and necrosis or subcutaneous tissue of the conjunctiva and nictitating membrane were not observed. The opacity of 4 scores lasted 48 hours in the cornea, but one animal was observed to be in the stage one recovery and the keratitis was diagnosed as corneal lesion which could be recovered. The response to light did not show up for 72 hours, but the histopathological examination found no abnormality in the iris. So, it was regarded as not irreversible lesion.

As a result of medicating iron dichloride to New Zealand White rabbits, the test substance was regarded as corrosive material because the lesion was not completely recovered for 21 days after the application.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Migrated information FeCl2 Criteria used for interpretation of results: EU
Conclusions:
The substance has been classified as R41 - Risk of serious damage to the eye according to the Dangersous Substances Directive (DSD). The substance has also been classified as category 1 (cause serious damage to the eye) according to the CLP regulations.
Executive summary:

To study eye irritation of iron dichloride (CAS No.7758-94-3) in three New Zealand White female rabbits, the subjects were dosed with the test substance. At 1 hour, and at Days 1, 2, 3, 7, 14, and 21, eye response and irritation in the cornea, iris and conjunctiva were evaluated and clinical signs were observed and body weight was measured.

The test results are as follows.

  • There was no mortality caused by the test substance during the test period.
  • Observing clinical signs, all the animals showed reduced mobility and two animals were observed to rub their eyes with frontal paws.
  • All the animals were observed to have a heavier body weight at Day 21 (after application) than at Day 0 (before application).
  • As a result of observing the applied region for local irritation at 1 hour, and Days 1, 2, 3, 7, 14, and 21, all the animals showed diffuse areas of opacity in the cornea at each observation time. An abnormality was found in the iris of all the animals, but histopathological signs were not observed. Severe redness, edema and excretion were observed in the conjunctiva of all the animals. Yellow pigmentation and exterior deformation were observed in the eyelid.
  • Hemosiderosis and lymphocyte infiltration in the submucosa of the conjunctiva were observed in the histopathological diagnosis.

Granulomatous inflammation was found in the conjunctiva and purulent keratitis was observed in the corneal stroma.

Hemosiderosis and severe lymphocyte infiltration were found in the dermis of the eyelid of one animal.

In conclusion, iron dichloride (CAS No. 7758-94-3) was evaluated as a corrosive substance that caused purulent inflammation in the conjunctiva and eyelid and granulomatous inflammation and purulent keratitis in the cornea, when applied to the eye of New Zealand White rabbits. Due to this the substance has been classified as R41 - Risk of serious damage to the eye according to the Dangerous Substances Directive (DSD). The substance has also been classified as category 1 (can cause serious damage to the eye) according to the CLP regulations EC No 1272/2008.