Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 1 – 11, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no deviations from standard test guidelines and no minor methodological deficiencies. This study is conducted according to an appropriate guideline and under the conditions of GLP and therefore the study is considered to be acceptable and to adequately satisfy both the guideline requirement and the regulatory requirement (Regulation (EC) No. 1907/2006; REACH) as a key study for this endpoint. In addition, this study is considered to be acceptable for classification and labelling in accordance with Regulation (EC) No. 1272/2008 (EU CLP).

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:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Manganese bis(dihydrogen phosphate)
EC Number:
242-520-2
EC Name:
Manganese bis(dihydrogen phosphate)
Cas Number:
18718-07-5
Molecular formula:
H3O4P.1/2Mn MnO8P2 H4MnO8P2
IUPAC Name:
manganese bis(dihydrogen phosphate)
Test material form:
solid: crystalline
Details on test material:
Composition: Manganese bis(dihydrogen phosphate) > 86%, CAS #18718-07-5
Physical Description: Pink, crystalline.
pH: 3.0-4.0 (1% solution in water)
Solubility: Soluble in water.
Stability: Test substance was expected to be stable for the duration of testing.
Expiration Date: September 8, 2014

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Received from Robinson Services, Inc. on September 26, 2012.
- Age at study initiation: Young adult.
- Sex: Female, nulliparous and non-pregnant.
- Housing: The animals were singly housed in suspended stainless steel caging with mesh floors, which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals (Natl. Res. Council, 2011). Litter paper was placed beneath the cage and was changed at least three times per week.
- Diet: Harlan Teklad Global High Fiber Rabbit Diet® #2031. A designated amount of the diet was available to each rabbit (approximately 150 grams/day).
- Water: Filtered tap water was supplied ad libitum by an automatic water dispensing system.
- Acclimation period: 5

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23ºC
- Humidity (%): 43-84% Humidity was above the targeted upper limit for 1 day during the study. A portable dehumidifier was used to lower the humidity levels during this time.
- Air changes (per hr): 12. Airflow measurements are evaluated regularly and the records are kept on file at Product Safety Labs.
- Photoperiod (hrs dark / hrs light): 12 hours light/dark

Contaminants: There were no known contaminants reasonably expected to be found in the food or water at levels which would have interfered with the results of this study. Analyses of the food and water are conducted regularly and the records are kept on file at Product Safety Labs.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: untreated eye served as control
Amount / concentration applied:
Instillation
Prior to instillation, 2-3 drops of ocular anesthetic (Tetracaine Hydrochloride Ophthalmic Solution, 0.5%) were placed into both the treated and control eye of each animal. One-tenth of a milliliter (0.10 grams) of the test substance was then instilled into the conjunctival sac of the right eye of each rabbit by pulling the lower lid away from the eyeball. The upper and lower lids were then gently held together for about one second before releasing to minimize loss of the test substance. The other eye of each rabbit remained untreated with the test substance and served as a control. The rabbits were then returned to their designated cages.
Duration of treatment / exposure:
10 days
Observation period (in vivo):
10 days post-instillation
Number of animals or in vitro replicates:
3
Details on study design:
Prior to test initiation, both eyes of a group of animals were examined using a white light source and a fluorescein dye procedure. One drop of 2% ophthalmic fluorescein sodium was instilled into both eyes of each rabbit. The eyes were rinsed with physiological saline (0.9% NaCl) approximately 30 seconds after instillation of the fluorescein and then evaluated for corneal damage using an ultraviolet light source. Prior to test substance instillation, the eyes were reexamined and scored for abnormalities according to the "Scale for Scoring Ocular Lesions". Three healthy animals without pre-existing ocular irritation were selected for test.


TOOL USED TO ASSESS SCORE: Ocular irritation was evaluated using a high-intensity white light (Mag Lite) in accordance with Draize et al at 1, 24, 48, and 72 hours and at 4, 7, and 10 days post-instillation. Individual scores were recorded for each animal. The fluorescein dye evaluation procedure was used in the treated eye at 24 hours and as needed at subsequent scoring intervals to evaluate the extent of corneal damage or to verify reversal of effects. In addition to observations of the cornea, iris and conjunctivae, any other observed lesions were noted. The average score for all rabbits at each scoring period was calculated to aid in data interpretation.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: Mean 24, 48 and 72 hours
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: Mean 24, 48 and 72 hours
Score:
0.66
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Irritation parameter:
iris score
Basis:
other: All animals tested
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
other: all animals tested
Time point:
other: Mean 24, 48 and 72 hours
Score:
3
Max. score:
3
Reversibility:
fully reversible within: 10 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: Mean 24, 48 and 72 hours
Score:
2.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: Mean 24, 48 and 72 hours
Score:
3
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: Mean 24, 48 and 72 hours
Score:
3.66
Max. score:
4
Reversibility:
fully reversible within: 10 days
Irritant / corrosive response data:
Individual eye irritation scores are presented in Table 1.

There was no iritis observed in any treated eye during this study. Within 24 hours of test substance instillation, two treated eyes exhibited corneal opacity and all three treated eyes exhibited "positive‟ conjunctivitis. The overall incidence and severity of irritation decreased gradually thereafter. An area of blanching on the conjunctivae was noted for one animal from 24 hours through Day 7. All animals were free of ocular irritation by Day 10 (study termination).
Other effects:
All animals appeared active and healthy during the study. Apart from the eye irritation noted below, there were no other signs of gross toxicity, adverse pharmacologic effects, or abnormal behavior.

Any other information on results incl. tables

Interpretation of Results

The numerical values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae were designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of cornea involved). For each tissue the score was calculated as follows:

Score for conjunctivae =         (A + B + C) x 2
Score for iris                            =         D x 5
Score for cornea                      =         (E x F) x 5

Using the numerical data obtained a modified version of the system (Modified Kay and Calandra Interpretation of Eye Irritation Test was used to classify the ocular irritancy potential of the test material. This was achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation were calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled classification of the eye irritancy potential of the test material.

If evidence of irreversible ocular damage is noted, the test material will be classified as corrosive to the eye.

 

Table 1             Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit 3401 Female

Rabbit 3402 Female

Rabbit 3403 Female

Hours

Days

Hours

Days

Hours

Days

Time After Treatment

1

24

48

72

4

7

10

1

24

48

72

4

7

10

1

24

48

72

4

7

10

CORNEA

 

 

 

 

 

 

 

 

 

E = Degree of Opacity

0

11

01

0

0

0

0

0

01

0

0

0

0

0

1

11

1

01

0

0

0

F = Area of Cornea Involved

4

1

4

4

4

4

4

4

4

4

4

4

4

4

1

1

1

4

4

4

4

Score (E x F) x 5

0

5

0

0

0

0

0

0

0

0

0

0

0

0

5

5

5

0

0

0

0

IRIS

 

 

 

 

 

 

 

 

 

D

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Score (D x 5)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

CONJUNCTIVAE

 

 

 

 

 

 

 

 

 

A = Redness

2

3

3

3

2

0

0

3

3

3

3

3

0

0

3

32

32

32

32

12

0

B = Chemosis

3

3

2

2

1

0

0

4

4

3

2

2

0

0

3

4

4

3

2

0

0

C = Discharge

2

2

2

1

1

0

0

2

3

2

2

1

0

0

2

3

3

2

1

0

0

Score (A + B + C) x 2

14

16

14

12

8

0

0

18

20

16

14

12

0

0

16

20

20

16

12

2

0

Total Score

14

21

14

12

8

0

0

18

20

16

14

12

0

0

21

25

25

16

12

2

0

 

12% ophthalmic fluorescein sodium used to evaluate the extent or verify the absence of corneal opacity.

2Area of blanching present on conjunctivae.

Applicant's summary and conclusion

Interpretation of results:
Category II
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In accordance with Regulation EC No. 1272/2008 on the Classification, Labeling, and Packaging of Substances and Mixtures, and based on the results of this study, Manganese bis(dihydrogen phosphate) classification: Category 2 (irritating to eye).