Registration Dossier

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:
2009-12-07 to 2009-12-17
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
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 the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

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:
Trimanganese tetraoxide
EC Number:
215-266-5
EC Name:
Trimanganese tetraoxide
Cas Number:
1317-35-7
Molecular formula:
Mn3O4
IUPAC Name:
dimanganese(3+) manganese(2+) tetraoxidandiide
Test material form:
solid: particulate/powder
Details on test material:
- Molecular formula of test material: Mn3O4
- Appearance: brown powder
- Physical state: Solid
- Impurities (identity and concentrations): K2O (1ppm), Na2O (3ppm), MgO (44ppm), CaO (14ppm), SiO2 (12ppm), S total (0.041%), H2O at 105 °C (0.42%)
- Composition of test material, percentage of components: Mn (71.2%)
- Lot/batch No.: 08100
- Storage condition of test material: 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: Harlan Laboratories U.K. Limited, Hillcrest, Belton, Loughborough, UK
- Age at study initiation: 12 to 20 weeks old
- Weight at study initiation: 2.13 to 2.66 kg
- Housing: Animals were housed individually in suspended cages.
- Diet : Free access to 2030 Teklad Global Rabbit diet (Harlan Teklad, Blackthorn, Bicester, Oxon, UK)
- Water : Free access to mains drinking water
- Acclimation period: A minimum of 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): At least 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hour cycle

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 54 mg
Duration of treatment / exposure:
Up to 72 hours
Observation period (in vivo):
Animals were assessed up to 72 hours after exposure (assessments were made at 1, 24, 48 and 72 hours post treatment)
Number of animals or in vitro replicates:
1 animal was initially treated, with a further two animals
Details on study design:
APPLICATION OF TEST SUBSTANCE:
Initially a single rabbit was treated with 54 mg of test material. The test material was administered into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment to prevent loss of test material and then released. The left eye served as a control and remained untreated. Immediately after administration of the test material an assessment of the initial pain reaction was made. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

SCORING SYSTEM:
Assessment of ocular damage/irritation was made 1, 24, 48 and 72 hours post dosing using the scoring system from Draize JH (1977) “ Dermal and Eye Toxicity Tests” In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p. 48 to 49. Any other ocular effects were also noted at this point along with any clinical signs of toxicity if present. Individual bodyweights were recorded on day 0 and 3. The numerical values corresponding to each animal, tissue and observation time were recorded. These scores were then assessed using a modified version of the system described by Kay JH and Calandra JC (1962), J. Soc. Cosmet. Chem. 13, 281-289. If evidence of irreversible ocular damage is noted, the test material will be classified as corrosive to the eye.


TOOL USED TO ASSESS SCORE: standard ophthalmoscope

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
Male
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Not applicable
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
Male
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Not applicable
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
Male
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Not applicable
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #1
Remarks:
Male
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #2
Remarks:
Male
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #3
Remarks:
Male
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within:
Remarks:
48 hours
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
Male
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
Male
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
Male
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritant / corrosive response data:
Moderate conjunctiva irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24-hr observation. All treated eyes appeared normal at the 48-hr observation

Any other information on results incl. tables

Measurement of pH

The pH of the test material was determined prior to commencement of the study and found to be as follows: 

Preparation

pH measurement

10% aqueous preparation of the test material

Immediately

After 10 minutes

After 20 minutes

6.8

8.8

8.5

Table 1: Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

68654 Male

68673 Male

68674 Male

IPR = 2

IPR = 2

IPR = 2

Time After Treatment

1 Hr

24 Hrs

48 Hrs

72 Hrs

1 Hr

24 Hrs

48 Hrs

72 Hrs

1 Hr

24 Hrs

48 Hrs

72 Hrs

Cornea

E = Degree of opacity

0

0

0

0

0

0

0

0

0

0

0

0

F = Area of Cornea Involved

0

0

0

0

0

0

0

0

0

0

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

0

0

0

0

Iris

D

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

Conjunctivae

A = Redness

2

1

0

0

2

1

0

0

2

1

0

0

B = Chemosis

2

1

0

0

2

1

0

0

2

1

0

0

C = Discharge

2Sf

1Sf

0Sf

0Sf

2Sf

1Sf

0Sf

0Sf

2Sf

1Sf

0Sf

0Sf

Score (A + B + C) x 2

12

6

0

0

12

6

0

0

12

6

0

0

Total Score

12

6

0

0

12

6

0

0

12

6

0

0

  IPR = Initial pain reaction

Sf = brown-coloured staining of the fur around treated eye

 

Table 2: Individual Total Scores and Group Mean Scores for Ocular Irritation

Rabbit Number and Sex

Individual Total Scores At:

1 Hour

24 Hours

48 Hours

72 Hours

68654 Male

12

6

0

0

68673 Male

12

6

0

0

68674 Male

12

6

0

0

Group Total

36

18

0

0

Group Mean Score

12.0

6.0

0.0

0.0

 

 

Table 3: Individual Bodyweights and Bodyweight Changes

Rabbit Number and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 3

68654 Male

2.66

2.75

0.09

68673 Male

2.13

2.32

0.19

68674 Male

2.21

2.27

0.06

 

Applicant's summary and conclusion

Interpretation of results:
other: Not classified according to EU criteria.
Conclusions:
Under the conditions of the study the test material was determined to be not irritating to eyes.
Executive summary:

The eye irritation potential of the test material was investigated in vivo in a study which was conducted under GLP conditions and in accordance with the standardised guidelines OECD 405 and EU Method B.5.

No corneal or iridial effects was noted during the study. Moderate conjuctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24 -hour observation. The test material was therefore concluded to be not irritating to eyes.