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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Remarks:
In accordance with Regulation (EU) 2016/863, preamble (7), adequate information from existing in vivo eye irritation study used to fulfil the standard information requirement point 8.2 Regulation (EC) No 1907/2006.
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991-02-11 to 1991-03-04
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report date:
1991

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
February 24, 1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
EEC Directive 84/449 EEC, EEC Publication No. L251, September 1984
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
dialuminium(3+) tetracalcium hexaoxidandiide phosphonate
EC Number:
915-069-0
Molecular formula:
not applicable
IUPAC Name:
dialuminium(3+) tetracalcium hexaoxidandiide phosphonate

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Selection of animals: Animals for test were selected from stock on the basis of background history. Each animal may have been used on a maximum of 3 previous occasions (in up to 2 skin irritation studies - one test each flank, and possibly 1 eye irritation study). No significant response has been noted on any previous occasion. Only eyes not previously treated were used for the test.
- Rational for selection: To minimise animal usage without compromising sudy integrity.
- Age at study initiation: approximately 20 weeks
- Weight at study initiation: 3106-3377 g
- Housing: Individually in cages with perforated floors equipped with an automatic drinking system (ITL, Bergen, The Netherlands)
- Diet: Standard laboratory rabbit diet (LKK-20, pellet diameter 4mm, Hope Farms, Woerden, The Netherlands) approximately 100 g per day
- Water: tap-water diluted with decalcified water ad libitum
- Acclimation period: At least five days before start of treatment under test conditions.


ENVIRONMENTAL CONDITIONS
- Temperature: 21°C
- Humidity: 55 %
- Air changes (per hr): 15
- Photoperiod: Lighting was 12 hours artificial fluorescent light and 12 hours dark per day.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The other eye of each animal remained untreated and served as the reference control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 62 +/- 2 mg (equivalent to 0.1 mL) per animal

VEHICLE
- no vehicle used
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
3
Details on study design:
TEST SUBSTANCE PREPARATION: The powdery test substance was instilled as delivered by the sponsor.

TREATMENT: On day 1, prior to test substance administration, the animals were physically examined and the eyes were found to be normal.

The test substance (approximately 62 mg (0.1 mL) per animal) was instilled in the conjunctival sac of one eye of each animal 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
- Washing (if done): Immediately after fluorescein examination on day 2, both eyes of all three animals were rinsed with approximately 50 mL tepid tap-water per eye, using a velocity of flow which did not affect the eyes, to remove residual test substance. However, not all test substance could be rinsed out because of adherence of test substance to the conjunctivae.
- Time after start of exposure: day 2 (24 hours after treatment)

SCORING SYSTEM: The test results were evaluated according to the EEC general classification and labelling requirements for dangerous substances (Annex VI of the EEC Council Directive 67/548/EEC as amended by Directive 83/467/EEC, Sep. 1983). Draize scores were calculated.

TOOL USED TO ASSESS SCORE:
- Immediately after the 24 hour observation, a solution of 2 % fluorescein 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 in all three animals on days 4 and 8. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.
- In case standard lighting was considered inadequate for observing minor effects, eye examinations were performed using a pocket flash light.
- In case of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, DC 20207 was used for additional control purposes.

OBSERVATIONS:
- Viability/Mortality: Daily, animals were also examined for signs of toxicity.
- Body weight: Day 1 of test (application day).
- Symptoms: The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation of the test substance.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.66
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: average score for time points 24/48/72 h; area of cornea affected: max. score 2
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
> 0 - < 1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: average score for time points 24/48/72 h Very slight corneal opacity, rated by study authors as score 0, but fluorescein positive 24 and 72 hours after instillation; area of cornea affected: max. score 2
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
> 0 - < 1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: average score for time points 24/48/72 h Very slight corneal opacity, rated by study authors as score 0, but fluorescein positive 24 and 72 hours after instillation; area of cornea affected: max. score 3
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.67
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.67
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.67
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
fully reversible within: 21 days
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
fully reversible within: 21 days
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
fully reversible within: 21 days
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.33
Max. score:
4
Reversibility:
fully reversible within: 21days
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: average score for time points 24/48/72 h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.33
Max. score:
4
Reversibility:
fully reversible within: 21 days
Remarks on result:
other: average score for time points 24/48/72 h
Irritant / corrosive response data:
Instillation of the substance resulted in reversible adverse effects on cornea, iris and conjunctivae in all three animals.

The opacity of the cornea was reversible within 7 days after exposure, the injection of the iris was reversible within 72 hours and the irritation of the conjunctivae was reversible within 21 days in all three animals.

There was no evidence of ocular corrosion.

No signs of systemic intoxication were observed during the study period. No animal died.

Any other information on results incl. tables

Table 1: Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point / Reversibility

Cornea opacity

Area of cornea affected

Iris

Conjunctivae

Chemosis

Discharge

Max. score: 4

Max. score: 4

Max. score: 2

Max. score: 3

Max. score: 4

Max. score: 3

60 min

01)/01)/01)

1/1/1

1/1/1

1/1/1

3/2/3

0/0/0

24 h

 01)/01)/01)

2/2/3

1/1/1

3/3/3

2/2/2

2/2/2

48 h

 1/01)/01)

1/2/2

1/1/1

3/3/3

2/2/2

3/3/3

72 h

 1/01)/01)

2/1/1

0/0/0

3/3/3

1/1/1

2/1/2

7 d

0/0/0

0/0/0

0/0/0

2/3/2

0/0/0

1/0/1

14 d

0/0/0

0/0/0

0/0/0

1/1/1

1/0/1

0/0/1

21 d

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

Average 24h, 48h, 72h

 0.66/01)/01)

1.67/1.67/2

 0.67/0.67/0.67

 3/3/3

2.33/2.33/2.33 

2.33/2/2.33

Reversibility*)

 c/c/c

 c/c/c

 c/c/c

 c/c/c

 c/c/c

c/c/c

Average time (d) for reversion

 7/7/7

 7/7/7

 3/3/3

 21/21/21

 21/7/21

14/7/21

1) Very slight corneal opacity, rated by study authors as score 0. Treatment of the eyes with 2 % fluorescein, 24 and 72 hours after test substance instillation revealed corneal epithelial damage in all three animals.

 *) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

The test substance was considered moderately irritating to the rabbit eye by a Kay and Calandra interpretation (J. Society of Cosmetic Chemists, Vol. B no.6, 1962) of the calculated maximum Draize score of 22.7 (maximum possible score 110).

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Remarks:
Criteria used for interpretation of results: Regulation (EC) No 1272/2008 (EU GHS)
Conclusions:
According to the criteria for classification and labelling of Regulation (EC) No 1272/2008, the test substance should be labelled as Irritating to eyes (Category 2).
Executive summary:

In a primary eye irritation study according to OECD Guideline No. 405, February 24, 1987 and EEC Directive 84/449/EEC, Part B.5, September 1984, 62 mg (0.1 mL) of substance "reaction mass of calcium hydrogen phosphonate and dialuminium tricalcium hexaoxide" (90 % a.i.) was instilled into the conjunctival sac of one eye of 3 young adult New Zealand white rabbits for 24 hours. Animals were observed for 21 days. Irritation was scored by the numerical scoring system according to Draize as given in the guidelines.

Instillation of the substance resulted in reversible adverse effects on cornea, iris and conjunctivae in all three animals.

The opacity of the cornea was reversible within 7 days after exposure, the injection of the iris was reversible within 72 hours and the irritation of the conjunctivae was reversible within 21 days in all three animals. There was no evidence of ocular corrosion.

In this study, substance "reaction mass of calcium hydrogen phosphonate and dialuminium tricalcium hexaoxide" (90 % a.i.) is an eye irritant.