Calcium acetylide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1995-02-15 to 1995-03-01

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Guideline study reliable with restrictions - The purity and stability of the test substance were not given. Minor deviations from the guideline with no effect on the results of the study: - According to the guideline, any effects other than ocular which are observed should be stated. It was not mentioned in this study if other than ocular effects were observed. - In this study only one animal was treated with the anaesthetic, but the other animals were not. This was considered to have no influence on the results of the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 1987-02-24

Deviations:

yes

Remarks:

see "rationale for reliability"

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Calciumdihydroxide

IUPAC Name:

Calciumdihydroxide

Constituent 2

Reference substance name:

Calcium dihydroxide

EC Number:

215-137-3

EC Name:

Calcium dihydroxide

Cas Number:

1305-62-0

Molecular formula:

CaH2O2

IUPAC Name:

calcium dihydroxide

Details on test material:

- Name of test material (as cited in study report): 150 g/l Calcium hydroxide (from Laboratoire Central Balthazard et Cotte)
- Physical state: Whitish liquid (suspension)
- Batch No.: 943638
- Storage condition of test material: At ambient temperature
- pH: The pH of the product, measured at the International Toxicology Centre, was about 9.
No further information on the test material was stated.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France
- Weight at study initiation: The animals had an average weight of 2.7 +/- 0.2 kg
- Housing: Polystyrene cage (35 X 55 X 32 cm or 48.2 X 58 X 36.5 cm) equipped with a trough and bottle.
- Diet (ad libitum): Food in the form of granules " Rabbit maintenance, Reference 112 C" (UAR, 91360 Villemoisson-sur-Orge,
France)
- Water (ad libitum): Drinking water
- Acclimation period: 5 days before the start of the study
ENVIRONMENTAL CONDITIONS
- Temperature: 18 +/- 3 °C
- Humidity: 30 to 70 %
- Photoperiod (hrs dark / hrs light): 12/12
No further information on the test animals was stated.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The lower eye lid was opened delicately and a single dose of 0.1 ml of the
product as such was introduced in the conjunctival cul-de-sac of the left eye of the 3 animals. The lower and upper lids were
maintained in contact for one second to prevent any loss of product. The right eye was not treated and served as control.
No further information on the amount/concentration applied was stated.

Observation period (in vivo):

The eyes were examined about 1 hour, 24, 48 and 72 hours after administration of the product. As there were signs of
persistent eye irritancy after 72 hours, the period of observation was extended by 21 days maximum (until Day 22), to
determine the progress of damage and reversibility or irreversibility thereof.

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
The eyes were not rinsed after administration of the product.
SCORING SYSTEM: Draize scoring system
All other damage observed was recorded.
TOOL USED TO ASSESS SCORE: 0.5 % aqueous fluorescein solution was used to evaluate the cornea (but not before 24
hours). If it was necessary the cornea was examined with an ultra-violet lamp. In the case of doubt as to the presence of
corneal opacity, the eye is subjected to UV examination (the areas of corneal impairment are distinguished by a very clear
fluorescence).
No further information on the study design was stated.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: at 48-hour observation

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: at 72-hour observation

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: observation time

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: observation time

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: at 72-hour observation

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: at Day 6

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: at Day 6

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: at Day 7

Results 10

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: at Day 7

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: at Day 8

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: at Day 7

Irritant / corrosive response data:

Very slight to pronounced chemosis and red blotch of the conjunctiva (score 1 to 3) were observed in all animals until day 5.
As of 24 hours and for 48 to 96 hours, clear or purulent watering was noted. Inflammation of the iris (score 1) was observed at
24 hours in 2 animals. Slight to very slight corneal opacity, accompanied by inflammation of the iris (score 1), was observed as
of days 2 to 6 in the third rabbit. No corneal opacity was noted in one rabbit, and only very slight opacity was observed 24 and
48 hours after treatment in the remaining animal. In the rabbit without damage to the cornea, total reversibility of the eye
reactions was observed on day 6. in the rabbit with very slight corneal opacitiy, focal necrosis of the palebral conjunctiva was
observed from day 6 to day 13. As of day 9, purulent watering reappeared, and as of day 12 to 17, the lids had to be forced
open for observation. As of day 6 and for the rest of the study, slight to serious chemosis (score 2 to 4) accompanied by a
slight to pronounced red blotch of the conjunctiva (score 2 or 3) was noted; the conjunctival reactions were more pronounced
as of day 12 to 17. In the third animal, focal necrosis of the palebral conjunctiva was observed from day 4 to the end of the
study. White spots appeared on the cornea on day 7, and were still observed on day 22. All the other eye reactions
disappeared on day 8.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Under the experimental conditions, the product, 150 g/l calcium hydroxide, is considered as an irritant to the eye in rabbits.
According to the EC Regulation No. 1272/2008 and subsequent regulations, the test item is classified as Category 1.

Executive summary:

In a primary eye irritation study performed according to OECD TG 405, 0.1 ml of calcium hydroxide (150 g/l) was instilled

into the conjunctival sac of the left eye of 3 male New Zealand White rabbits. Eyes were not rinsed after administration of the test item. Animals were observed for 21 days. Irritation was scored by the method of Draize.

Very slight to pronounced chemosis and red blotch of the conjunctiva were observed in all animals until day 5.

As of 24 hours and for 48 to 96 hours, clear or purulent watering was noted. Inflammation of the iris was observed at 24 hours in 2 animals. Slight to very slight corneal opacity, accompanied by inflammation of the iris, was observed as of days 2 to 6 in the third rabbit. In the rabbit without damage to the cornea, total reversibility of the eye

reactions was observed on day 6. In the rabbit with very slight corneal opacitiy, focal necrosis of the palebral conjunctiva was observed from day 6 to day 13. As of day 9, purulent watering reappeared, and as of day 12 to 17, the lids had to be forced open for observation. As of day 6 and for the rest of the study, slight to serious chemosis accompanied by a slight to pronounced red blotch of the conjunctiva was noted; the conjunctival reactions were more pronounced as of day 12 to 17. In the third animal, focal necrosis of the palebral conjunctiva was observed from day 4 to the end of the study. White spots appeared on the cornea on day 7, and were still observed on day 22. All the other eye reactions disappeared on day 8.

Under the experimental conditions, calcium hydroxide, is considered as an irritant to the eye in rabbits.

According to Regulation (EC) No. 1272/2008 and subsequent regulations, the test item is classified as Eye Dam. Category 1.