Aluminium lanthanum trioxide

Endpoint summary

Description of key information

Key studies demonstrate a lack of irritation/corrosion potential: 

A Bovine Corneal Opacity and Permeability Test for identifying ocular corrosives and severe irritants according to OECD 437 (Duschl, 2017a) was performed with aluminium lanthanum trioxide, and results indicate that it is not corrosive to the eye. A Reconstructed Human Cornea-like Epithelium (RhCE) Test for identifying eye irritants and corrosives according to OECD TG 437 (Duschl, 2017b) was performed with aluminium lanthanum trioxide, and results indicate that it is not irritant to the eye.

Data regrading skin-irritating properties are read-across based on grouping of substances, i.e.aluminium oxide, lanthanum oxide and aluminium lanthanum oxide, based on inertness and similar solubility or lack thereof. In vivo skin irritation / corrosion tests similar to OECD 404 (Lambert, 190) performed with lanthanum oxide and aluminium oxide further indicate that aluminium oxide and lanthanum oxide do not possess a skin irritation potential. Based on read-across (see category approach), it is assumed that aluminium lanthanum trioxide does also not cause skin irritation.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions

Justification for type of information:

The solubility of aluminium lanthanum trioxide (AlLaO3) is low since dissolution of AlLaO3 in water resulted in La concentrations < 0.01 mg/L and Al concentrations < 0.03 mg/L after 34 days. The dissolution of AlLaO3 in water results in Al (3+) (=Al(H2O)6 (3+)) ions and La (3+) ions. Thus, the toxicological moieties of concern are aluminium and lanthanum ions. Thus, in the assessment of toxicity, data available for different aluminium and lanthanum substances are read-across since aluminium and lanthanum ions determine the toxicological potential of aluminium lanthanum trioxide, i.e. are the common toxicological moieties of concern. The effects of the substance aluminium lanthanum trioxide with regard to skin irritation are predicted to be equal to the effects of Al (3+) ions and La (3+) ions.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

: Longer duration of exposure (24 versus 4 hours); no detail on pH of test substance; limited detail on characteristics of the test substance.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
Age: not reported
Sex: not reported
Weight: not reported

Environmental/Housing conditions:
No information provided.

Acclimation: No information was provided.

Feeding conditions:
This information was not provided.

Type of coverage:

other: wrapped with binders of rubber dam

Preparation of test site:

other: intact and abraded skin

Vehicle:

water

Controls:

no

Amount / concentration applied:

TEST MATERIAL
-Concentrations: 0.5g in 1.0mL = 500g/L (50%)
-Volume: 1.0 mL
-Area: the substance was under a 1 inch square patch.

Duration of treatment / exposure:

A single, 24 hour dermal application was made.

Observation period:

Observations were made at 24 and 72 hours.

Number of animals:

Three animals with intact skin and three animals with abraded skin

Details on study design:

One-inch square gauze patches were applied to pre-moistened, clipped intact or abraded skin on the back of the animals, wrapped with binders of rubber dam and immobilised for the exposure period.

The scoring system used was that of Draize (1959) and was provided in the report:
Evaluation of Skin reactions
Erythema and eschar formation:
No erythema, Score = 0
Very slight erythema (barely perceptible), Score =1
Well-defined erythema, Score=2
Moderate to severe erythema, Score=3
Severe erythema (beet redness) to slight eschar formation (injuries in depth), Score =4

Edema formation:
No edema, Score=0
Very slight edema (barely perceptible), Score=1
Slight edema (edges of area will defined by definite raising), Score=2
Moderate edema (raised approximately 1.0mm)
Severe edema (raised more than 1.0mm extending beyond the area of exposure), Score=4

-“Values for erythema and eschar formation at 24 hours and 72 hours for intact skin animals will be added to the values on abraded skin animals at 24 and 72 hours. Similarly, the values for edema formation at 24 hours and at 72 hours for the intact and abraded skin animals will be added. The primary irritant score is the total of the eight values divided by four. A primary irritant is one which results in a score of five or more as tested by this method.”

The pH of the test substance was not measured.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3/3

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3/3

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3/3

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

mean

Remarks:

3/3

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Remarks:

3/3

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Remarks:

3/3

Time point:

48 h

Remarks on result:

not measured/tested

Other effects:

Clinical signs of toxicity were not reported.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The results from the study are negative and would not lead to classification.

Executive summary:

A study conducted at Hazleton Laboratories Inc. for the Cabot Corporation investigated dermal irritation effects of ALON, a fumed alumina produced by the chemical hydrolysis of aluminium chloride, in New Zealand White rabbits. 0.5g of the substance in 1.0mL of distilled water was applied to pre-moistened skin on the backs of 6 rabbits under gauze patches. Three of the rabbits had abraded skin and three intact skin. The patches remained place for 24 hours. Lesions were scored at 24 hours and at 72 hours using the scoring criteria of Draize (1959). The report lacks detail on animal husbandry and signs of toxicity. Further details of the test substance characteristics were obtained from the study sponsor. The results showed slight erythema in all three rabbits with abraded skin at 24 hours. The erythema had resolved by 72 hours. No erythema was observed in the animals with intact skin. No animal exhibited edema. The overall primary irritation score from the study (0.25) would not lead to classification.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions

Justification for type of information:

The solubility of aluminium lanthanum trioxide (AlLaO3) is low since dissolution of AlLaO3 in water resulted in La concentrations < 0.01 mg/L and Al concentrations < 0.03 mg/L after 34 days. The dissolution of AlLaO3 in water results in Al (3+) (=Al(H2O)6 (3+)) ions and La (3+) ions. Thus, the toxicological moieties of concern are aluminium and lanthanum ions. Thus, in the assessment of toxicity, data available for different aluminium and lanthanum substances are read-across since aluminium and lanthanum ions determine the toxicological potential of aluminium lanthanum trioxide, i.e. are the common toxicological moieties of concern. The effects of the substance aluminium lanthanum trioxide with regard to skin irritation are predicted to be equal to the effects of Al (3+) ions and La (3+) ions.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

: longer duration of exposure (24 versus 4 hours); limited details on test material and test conditions

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

semiocclusive

Preparation of test site:

other: intact or abraded skin

Vehicle:

not specified

Controls:

no

Amount / concentration applied:

0.5 g of the solid test substance was brought into contact with the intact or abraded skin under a surgical patch measuring 1 inch Χ 1 inch.

Duration of treatment / exposure:

24 hours

Observation period:

24 hours and 72 hours after the substance application

Number of animals:

6 rabbits were treated on intact skin and 6 on abraded skin.

Details on study design:

TEST SITE
- Area of exposure: the hair was removed from the backs of the animals with an electric clipper avoiding abrasions. The test material was placed under a surgical patch 1 inch Χ 1 inch (2.5 cm Χ 2.5 cm) on intact or abraded skin.
- Type of wrap if used: surgical patch. The patches were fixed to the application site with adhesive tape. The entire trunk of the animals was wrapped with an impervious material to keep the patches in place.


SCORING SYSTEM: Draize scoring system


The abrasions were “minor incisions through the stratum corneum, but not sufficiently deep to disturb the derma or to produce bleeding.”

The patches were fixed to the application site with adhesive tape. The entire trunk of the animals was wrapped with an impervious material to keep the patches in place

After exposure for 24 hours, the patches and the test material were removed (no information was provided on washing of the skin after patch removal.)

Skin lesions were evaluated at the time of patch and material removal and 48 hours later (24 and 72 hours after the application)

Scoring systems applied:
-24 hours after application -method of Draize (J. Pharmocol. 82 (1944) 377-390).
-72 hours after application – CIVO grading system (because the Draize scoring system does not describe scaliness and necrosis)

Evaluation of skin reactions (Draize, 1944) Value
A. Erythema and eschar formation

No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

B. Edema formation

No edema 0
Very slight edema (barely perceptible) 1
Slight edema (edges of area well defined by definite raising) 2
Moderate edema (raised approximately 1 mm) 3
Severe edema (raised more than 1 mm and extending beyond the area of exposure) 4

Evaluation of skin reactions (CIVO) Value
No reaction at all 0
Very slight scaliness 1
Distinct scaliness or very slight incrustation 2
Distinct incrustation 3
Severe incrustation 4

Irritation parameter:

erythema score

Basis:

mean

Remarks:

6/6

Time point:

24/48/72 h

Score:

0.166

Max. score:

4

Reversibility:

other: not reported

Irritation parameter:

edema score

Basis:

mean

Remarks:

6/6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Overall, the test material caused slight erythema in 2 of the 12 rabbits (one - 24 hours after application, and the other - 72 hours after application).

Other effects:

Clinical signs of toxicity were not reported

Individual and average skin reaction scores

1)     Intact skin

--------------------------------------------------------------------------------------------

Rabbit number                           24 hour          72 hour

                                                   A-B                                A-B

---------------------------------------------------------------------------------------------

8416                                             0-0                                 0-0

8417                                             0-0                                 0-0

8418                                             0-0                                 1-0

8419                                             0-0                                 0-0

8420                                             1-0                                 0-0

8421                                             0-0                                 0-0

--------------------------------------------------------------------------------------------

Average                                         0.2                                 0.2

--------------------------------------------------------------------------------------------

 

2)     Abraded skin

--------------------------------------------------------------------------------------------

Rabbit number                           24 hour                             72 hour

                                                   A-B                               A-B

---------------------------------------------------------------------------------------------

8410                                             0-0                                0-0

8411                                             0-0                                0-0

8412                                             0-0                                0-0

8413                                             0-0                                0-0

8414                                             0-0                                0-0

8415                                             0-0                                0-0

--------------------------------------------------------------------------------------------

Average                                         0.0                                 0.0

--------------------------------------------------------------------------------------------

A – erythema; B – edema

 

Overall, the test material caused slight erythema in 2 of the 12 rabbits (one - 24 hours after application, and the other - 72 hours after application). Conclusion: Due to the slight character of the observed effects the test substance does not have to be classified according to the criteria of the DSD and the CLP regulation.

Interpretation of results:

not irritating

Remarks:

Migrated information The applicant’s interpretation of the results are in accordance with the conclusions of the study authors - “AK 43/79 [the code of the test material] caused slight erythema in 2/12 rabbits and is, therefore, not considered to be a primary skin irritant.” Criteria used for interpretation of results: EU

Conclusions:

The observed slight erythema effects do not lead to a classification for the test item. Therefore AK 43/79 is not considered to be a primary skin irritant.

Executive summary:

This study conducted for Degussa corporation investigated dermal irritation effects of aluminium oxide in New Zealand White albino rabbits. 0.5 g of the test substance was placed on the intact or abraded skin of the rabbits under a surgical patch measuring 2.5 Χ 2.5 cm. Six rabbits had intact skin and six rabbits abraded skin. The exposure continued for 24 hours (a 4 hour exposure is recommended by OECD 404), and skin lesions were scored immediately after patch removal and 48 hours later (24 and 72 hours after the application of the test material). Two sets of scoring criteria were used: Draize (1944) - 24 hours after the material application and CIVO - 72 hours after the application. 

Insufficient information was provided on the test material and in what form it was applied to the skin. No information was provided on animal husbandry. Slight erythema was observed on the intact skin of 2 animals (one 24 hours after the substance application and the other 72 hours after application). No signs of skin irritation were observed on the abraded skin at any time. The overall primary irritation score from the study (0.2) would not lead to classification.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions

Justification for type of information:

The solubility of aluminium lanthanum trioxide (AlLaO3) is low since dissolution of AlLaO3 in water resulted in La concentrations < 0.01 mg/L and Al concentrations < 0.03 mg/L after 34 days. The dissolution of AlLaO3 in water results in Al (3+) (=Al(H2O)6 (3+)) ions and La (3+) ions. Thus, the toxicological moieties of concern are aluminium and lanthanum ions. Thus, in the assessment of toxicity, data available for different aluminium and lanthanum substances are read-across since aluminium and lanthanum ions determine the toxicological potential of aluminium lanthanum trioxide, i.e. are the common toxicological moieties of concern. The effects of the substance aluminium lanthanum trioxide with regard to skin irritation are predicted to be equal to the effects of Al (3+) ions and La (3+) ions.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

: longer duration of exposure (24 versus 4 hours); limited details on test material and test conditions

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Environmental/Housing conditions: No information

Acclimation: No information.

Feeding conditions: No information.

Type of coverage:

semiocclusive

Preparation of test site:

other: intact or abraded skin

Vehicle:

not specified

Controls:

no

Amount / concentration applied:

0.5 g of the solid test substance was brought into contact with the intact or abraded skin under a surgical patch measuring 1 inch Χ 1 inch.

Duration of treatment / exposure:

24 hours

Observation period:

24 hours and 72 hours after the substance application

Number of animals:

6 rabbits were treated on intact skin and 6 on abraded skin.

Details on study design:

TEST SITE
- Area of exposure: the hair was removed from the backs of the animals with an electric clipper avoiding abrasions. The test material was placed under a surgical patch 1 inch Χ 1 inch (2.5 cm Χ 2.5 cm) on the intact or abraded skin.
- Type of wrap if used: surgical patch.The patches were fixed to the application site with adhesive tape. The entire trunk of the animals was wrapped with an impervious material to keep the patches in place.


SCORING SYSTEM: Draize scoring system

 
The abrasions were “minor incisions through the stratum corneum, but not sufficiently deep to disturb the derma or to produce bleeding.”
 
The patches were fixed to the application site with adhesive tape. The entire trunk of the animals was wrapped with an impervious material to keep the patches in place and “to retard evaporation of volatile substances.”
 
After exposure for 24 hours, the patches and the test material were removed (no information on the mode of removal)
 
Skin lesions were evaluated at the time of patch and material removal and 48 hours later (24 and 72 hours after the application)
 
Scoring systems applied:
-         24 hours after application -method of Draize (J. Pharmocol. 82 (1944) 377-390).
-         72 hours after application – CIVO grading system (because the Draize scoring system does not describe scaliness and necrosis)
 
Evaluation of skin reactions (Draize, 1944)                                      Value
A.     Erythema and eschar formation
 
No erythema                                                                     0
Very slight erythema (barely perceptible)                                1
Well-defined erythema                                                             2
Moderate to severe erythema                                                  3
Severe erythema (beet redness) to slight eschar formation (injuries in depth)       4
 
B.     Edema formation
 
No edema                                                                                 0
Very slight edema (barely perceptible)                                     1
Slight edema (edges of area well defined by definite raising)  2
Moderate edema (raised approximately 1 mm)                        3
Severe edema (raised more than 1 mm and extending beyond the area of exposure)   4
 
Evaluation of skin reactions (CIVO)                                                Value
No reaction at all                                                                            0
Very slight scaliness                                                                       1
Distinct scaliness or very slight incrustation                                   2
Distinct incrustation                                                                        3
Severe incrustation                                                                         4

Irritation parameter:

erythema score

Basis:

mean

Remarks:

6/6

Time point:

24/48/72 h

Score:

0.22

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

edema score

Basis:

mean

Remarks:

6/6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Other effects:

Clinical signs of toxicity were not reported

Individual and average skin reaction scores

1)     Intact skin

--------------------------------------------------------------------------------------------

Rabbit number                           24 hour                           72 hour

                                                   A-B                               A-B

---------------------------------------------------------------------------------------------

8416                                             0-0                                0-0

8417                                             0-0                                0-0

8418                                             1-0                                0-0

8419                                             0-0                                0-0

8420                                             1-0                                0-0

8421                                             0-0                                0-0

--------------------------------------------------------------------------------------------

Average                                        0.3                                 0.0

--------------------------------------------------------------------------------------------

 

2)     Abraded skin

--------------------------------------------------------------------------------------------

Rabbit number                           24 hour                           72 hour

                                                   A-B                               A-B

---------------------------------------------------------------------------------------------

8410                                             0-0                                0-0

8411                                             0-0                                0-0

8412                                             0-0                                0-0

8413                                             0-0                                0-0

8414                                             0-0                                0-0

8415                                             0-0                                0-0

--------------------------------------------------------------------------------------------

Average                                         0.0                                0.0

--------------------------------------------------------------------------------------------

A – erythema; B – edema

 

Overall, the test material caused slight erythema in 2 of the 12 rabbits 24 hours after application. No sign of skin irritation was observed at 72 hours.

Interpretation of results:

not irritating

Remarks:

Migrated information “AK 44/79 [the code of the test material] caused slight erythema in 2/12 rabbits and is, therefore, not considered to be a primary skin irritant.” Criteria used for interpretation of results: EU

Conclusions:

The test item, AK 44/79 caused slight erythema in 2/12 rabbits. The observed effects do not lead to a classification. AK 44/79 is, therefore, not considered to be a primary skin irritant.”

Executive summary:

This study conducted for Degussa corporation investigated dermal irritation effects of aluminium oxide inNew ZealandWhite albino rabbits.0.5 g of the test substance was placed on the intact or abraded skin of the rabbits under a surgical patch measuring 2.5 Χ 2.5 cm. Six rabbits had intact skin and six rabbits had abraded skin. The exposure continued for 24 hours (4 hour exposure is recommended by OECD 404), and skin lesions were scored immediately after patch removal and 48 later (24 and 72 hours after the application of the test material). Two sets of scoring criteria were used: Draize (1944) - 24 hours after the material application and CIVO - 72 hours after the application.

 

Insufficient information wss provided on the test material and application of the test material. No information was provided on animal husbandry. Slight erythema was observed on theintactskin of 2 animals 24 hours after the substance application. No signs of skin irritation were seen at 72 hours. No signs of skin irritation were observed on theabradedskin at any time. The overall primary irritation score from the study (0.3 at 24 hours) would not lead to classification.

Reference 4

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Justification for type of information:

The solubility of aluminium lanthanum trioxide (AlLaO3) is low since dissolution of AlLaO3 in water resulted in La concentrations < 0.01 mg/L and Al concentrations < 0.03 mg/L after 34 days. The dissolution of AlLaO3 in water results in Al (3+) (=Al(H2O)6 (3+)) ions and La (3+) ions. Thus, the toxicological moieties of concern are aluminium and lanthanum ions. Thus, in the assessment of toxicity, data available for different aluminium and lanthanum substances are read-across since aluminium and lanthanum ions determine the toxicological potential of aluminium lanthanum trioxide, i.e. are the common toxicological moieties of concern. The effects of the substance aluminium lanthanum trioxide with regard to skin irritation are predicted to be equal to the effects of Al (3+) ions and La (3+) ions.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1981-05-12

Deviations:

yes

Remarks:

purity of test item, vehicle & housing conditions missing; observations at 30-60 min. & 48 hrs after exposure missing; observations of any toxic effects besides skin irritation missing; individual animal data missing

GLP compliance:

no

Specific details on test material used for the study:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

not specified

Type of coverage:

occlusive

Preparation of test site:

other: intact and abraded skin sites

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g of test item

Duration of treatment / exposure:

24 hours

Observation period:

72 hours

Number of animals:

6 rabbits

Details on study design:

TEST SITE
- Area of exposure: the test item was applied to one intact and one abraded skin site (approx. 2.5 cm²) of six rabbits and occluded for 24 hours.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): patches were removed and the test sites wiped
- Time after start of exposure: 24 hours

OBSERVATION TIME POINTS
Sites were evaluated 24 and 72 hours after initial exposure.

SCORING SYSTEM: method of Draize

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #4

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #4

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #5

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #5

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #6

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #6

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

48 h

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #1

Time point:

72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

48 h

Remarks on result:

not measured/tested

Irritant / corrosive response data:

There were no signs of dermal irritation.

Interpretation of results:

GHS criteria not met

Conclusions:

The substance is not irritating to the skin based on an in vivo study similar to OECD TG 404.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-09-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

2013-07-26

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Version / remarks:

2010

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Bovine Corneal Opacity and Permeability (BCOP) Assay, SOP of Microbiological Associates Ltd., UK, Procedure Details, April 1997

Version / remarks:

April 1997

GLP compliance:

yes (incl. QA statement)

Remarks:

signed 2015-09-14

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: abattoir (Schlachthof Aschaffenburg, 63739 Aschaffenburg, Germany)
- Characteristics of donor animals: at least 9 month old donor cattle
- Storage, temperature and transport conditions of ocular tissue: isolated eyes were transported to the laboratory in Hank’s Buffered Salt Solution (HBSS) supplemented with streptomycin / penicillin at ambient temperature.
- Time interval prior to initiating testing: corneae were isolated and used on the same day after delivery of the eyes

Vehicle:

physiological saline

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration: 20 % suspension (w/v) in vehicle

Duration of treatment / exposure:

240 minutes

Observation period (in vivo):

not applicable

Duration of post- treatment incubation (in vitro):

not required

Number of animals or in vitro replicates:

Number of bovine corneae per dose:
Test item: triplicates
Negative control: triplicates
Positive control: triplicates

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
- all eyes were examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity and scratches were discarded.
- cornea was removed from the eye.
- each cornea was mounted in a cornea holder according to the description given in OECD guideline 437, which consists of anterior and posterior
compartments, which interface with the epithelial and endothelial sides of the cornea, respectively. Both compartments of the holder were filled with incubation medium (MEM*, equivalent to EMEM).
- for equilibration, the corneae in the holder were incubated in a vertical position for about one hour at 32 ± 1 °C in a water-bath.

*MEM supplemented with sodium bicarbonate, L-glutamine, penicillin/streptomycin, and 1% fetal calf serum

QUALITY CHECK OF THE ISOLATED CORNEAS
- at the end of the equilibration period, the basal opacity was determined (t0).
- each cornea with a value of the basal opacity > 7 was discarded.

APPLICATION DOSE / EXPOSURE TIME / REMOVAL OF TEST SUBSTANCE
- the anterior compartment received the test item suspension or negative or positive control at a volume of 0.75 mL each on the surface of the corneae.
- corneae were incubated in a horizontal position at 32 ± 1 °C in the water-bath (incubation time: 240 minutes).
- after the incubation period, the test item or control items, respectively, were rinsed off from the application side with saline
- fresh incubation medium was added into the anterior compartment and opacity was measured (t240).
- permeability of the cornea was determined

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: the opacitometer (OP_KiT opacitometer (Electro Design)) was calibrated and the opacity of each of the corneae was determined by reading each holder placed in the photoreceptor compartment for treated cornea.
Evaluation of opacity:
- the change of opacity value of each treated cornea or positive and negative control corneae is calculated by subtracting the initial basal opacity from the post treatment opacity reading (t240 – t0), for each individual cornea.
- the average change in opacity of the negative control corneae is calculated and this value is subtracted from the change in opacity of each treated cornea or positive control to obtain a corrected opacity.

- Corneal permeability: passage of sodium fluorescein dye measured with the aid of spectrophotometer (Versamax® Molecular Devices) (OD490).
- after the final opacity measurement was performed, the incubation medium was removed from the anterior compartment and replaced by a 0.5% (w/v) sodium fluorescein solution in HBSS.
- corneae were incubated again in a horizontal position for 90 ± 10 minutes in a water-bath at 32 ± 1 °C.
- incubation medium from the posterior compartment were removed, well mixed and transferred into a 96 well plate and the optical density at 490 nm (OD490) was determined with a spectrophotometer.
Evaluation permeability:
- the corrected OD490 value of each cornea treated with positive control and test item is calculated by subtracting the average negative control cornea value from the original
permeability value for each cornea.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
The following formula was used to determine the IVIS of the negative control:
IVIS = opacity value + (15 x OD490 value)
The following formula was used to determine the IVIS of the positive control and the test item:
IVIS = (opacity value – opacity value mean negative control) + (15 x corrected OD490 value)
The mean IVIS value of each treated group was calculated from the IVIS values of each individual treatment and positive control cornea.
Depending on the score obtained, the test item was classified into categories according to OECD guideline 437 (table 1 in the field "Any other information on materials and methods incl. tables" below).

ACCEPTANCE CRITERIA:
The test was acceptable if
- the positive control gives an IVIS that falls within two standard deviations of the current historical mean (updated every three months), and if
- the negative control responses result in opacity and permeability values that are less than the established upper limits for background opacity and permeability values for bovine corneae treated with the respective negative control.

Irritation parameter:

in vitro irritation score

Remarks:

(mean)

Value:

17.84

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- with the negative control (saline) neither an increase of opacity nor permeability of the corneae could be observed (mean IVIS = 1.48).
- distinct opacity of the corneae leading to a mean IVIS of 110.15 was caused after treatment with the positive control (10% (w/v) Benzalkonium chloride in saline) corresponding to the classification Irreversible effects on the eye / serious eye damag (CLP (Cat 1)).

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: opacity and permeability of the negative control are less than the respective established upper limits for background opacity and
permeability.
- Acceptance criteria met for positive control: positive control falls within two standard deviations of the current historical mean.

Please refer to the field "Any other information on results incl. tables" below.

Table 1: Results after 240 minutes incubation time

Test Group	Opacity value = Difference (t240-t0) of Opacity		Permeability at 490 nm (OD490)		IVIS	Mean IVIS
		Mean		Mean
Negative Control	0	0.33	0.079	0.077	1.19	1.48
	1		0.076		2.14
	0		0.075		1.13
Positive Control	117.67*		0.003*		117.72	110.15
	104.67*		0.017*		104.93
	107.67*		0.009*		107.81
Aluminium lanthanum trioxide	20.67*		0.114*		22.38	17.84
	10.67*		0.190*		13.52
	14.67*		0.197*		17.63

*corrected values

Table 2: Historical data

	Positive Control	Negative Control
Mean IVIS	122.01	1.32
Standard Deviation IVIS	15.51	0.33
Range of IVIS	98.49 – 167.85	0.73 – 2.04
Mean Opacity t240min	120.03	0.17
Standard Deviation Opacity t240min	18.45	0.25
Range of Opacity t240min	88.67 – 183.00	-0.33 – 0.67
Mean Permeability	0.147	0.184
Standard Deviation Permeability	0.257	0.281
Range of Permeability	0.000 – 1.438	0.055 – 1.100

Values of 32 studies with solid test items performed from February 2015 (calendar week 7) until October 2016 (calendar week 40)

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, based on a valid OECD 437 test and under the experimental conditions reported, aluminium lanthanum trioxide does not meet classification criteria of irreversible effects on the eye / serious eye damage (CLP (Cat 1).

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-02-23 to 2017-03-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

2015-07-28

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol: EpiOcular™ Eye Irritation Test (OCL-200-EIT) for the prediction of acute ocular irritation of chemicals; for use with MatTek Corporation’s Reconstructed Human EpiOcular™ Model

Version / remarks:

2015-06-29

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

signed 2015-09-14

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature

Details on test animals or tissues and environmental conditions:

JUSTIFICATION OF THE TEST METHOD
In a prevalidation study performed by Avon Products Inc. and MatTek Corporation, the in vitro eye irritation using the human cornea model EpiOcular™ and measurement of cell viability
by dehydrogenase conversion of MTT into a blue formazan salt has turned out to be a sufficiently promising predictor for eye irritancy potential.
The EpiOcular ™ Eye Irritation Test (EIT) has been validated by the European Union Reference laboratory for Alternatives to Animal Testing (EURL ECVAM) and cosmetics Europe between 2008 and 2013.

HUMAN RECONSTRUCTED CORNEA MODEL
- Model used: human cornea model EpiOcular™ (source: MatTek Corporation, 82105 Bratislava, Slovakia)
The EpiOcular™ tissue consists of normal, human-derived epidermal keratinocytes that have been cultured to form a stratified squamous epithelium morphologically similar to that found in the human cornea. It consists of highly organized basal cells which progressively flatten out as the apical surface of the tissue is approached, analogous to the normal in vivo corneal epithelium. The EpiOcular™ RhCE tissue construct consists of at least 3 viable layers of cells and a nonkeratinized surface with a cornea-like structure analogous to that found in vivo.
- Tissue lot number: 23771

FUNCTIONAL MODEL CONDITIONS
- Viability: tissues pass analysis for tissue viability
- Barrier function: tissues pass analysis for tissue functionality
- Contamination: absence of bacteria, yeast, and other fungi (long term antibiotic, antimycotic free culture) as well as absence of HIV1- virus, Hepatitis B virus, and Hepatitis C virus
Please also refer to the field "Attached background material" below.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): approx. 50 mg

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): 50 µL

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 50 µL

Duration of treatment / exposure:

6 hours

Observation period (in vivo):

not applicable

Duration of post- treatment incubation (in vitro):

18 hours

Number of animals or in vitro replicates:

duplicates

Details on study design:

ASSESSMENT OF DIRECT MTT REDUCTION
The test item was evaluated for its potential to directly reduce MTT. For this purpose approx. 50 mg of the test item were added to 1 mL MTT solution (1 mg/mL; in DMEM) in a glass tube, and the mixture was incubated in the dark at standard culture conditions for three hours. A control (50 μL of deionised water in 1 mL of 1.0 mg/mL MTT solution) was run concurrently. If the MTT solution colour would turn blue/purple, the test item was presumably to have reduced the MTT.

ASSESSMENT OF COLOURED OR STAINING MATERIALS
The test item was checked for its colourant properties, which might interfere with the quantitative photometric MTT measurement.
Since the test item was non-coloured additional tests had to be performed to assess if it became coloured after contact with water or isopropanol. For this purpose, approx.
50 mg of the test item were added to 1.0 mL of water and to 2 mL isopropanol in a glass tube, respectively. The water mixture was incubated in the dark at standard culture conditions for 1 hour, whereas the isopropanol mixture was incubated for 3 hours at room temperature.

PRE-EXPERIMENT CONDITIONS
- on day of receipt of the EpiOcular™ tissues, a equilibration step was started (15 minutes at room temperature in the shipping container).
- after the equilibration step, each tissue was inspected for air bubbles between the agarose gel and insert.
- insert was transferred aseptically into the 6-well plates and pre-incubated at standard culture conditions for one hour in Assay Medium.
- after one hour, the Assay Medium was replaced by fresh Assay Medium at 37 °C
- EpiOcular™ tissues were incubated at standard culture conditions overnight (about 18 hours).

EXPERIMENTAL PERFORMANCE
- after the overnight incubation, the tissues were pre-wetted with 20 μL of Ca++& Mg++free-DPBS and were incubated at standard culture conditions for 30 minutes.
- after the 30 minute period, the test and control item were applied topically onto the EpiOcular™ tissues.
- tissues were incubated at standard culture for 6 hours.
- after the 6 hours, the test item was removed by rinsing the tissues several times with Ca++& Mg++-free DPBS (brought to room temperature).
- it was not possible to remove the visible test item completely, but no further rinsing was done.
- after rinsing, the tissues were transferred to and immersed in assay medium (room temperature) in a 12-well plate for 25 minutes immersion incubation at room temperature (to remove any test item adsorbed by the tissue).
- next, each insert was removed from the assay medium, the medium was decanted off the tissue, and the insert was blotted and transferred into the appropriate well of a 6-well plate containing warm assay medium.
- tissues were incubated for about 18 hours at standard culture conditions (post-treatment incubation).

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- at the end of the post-treatment incubation, each insert was removed from the plate and blotted.
- tissues were placed into a 24-well plate containing 0.3 mL of MTT solution.
- plate was incubated for 180 minutes at standard culture conditions.
- next, inserts were removed from the 24-well plate, bottom of the insert was blotted, and then transferred into 6-well plate containing 2 mL isopropanol in each well.
- plates were sealed with parafilm or a standard plate sealer, and were extracted (shaken for 2.5 hours at room temperature).
- tissues were not pierced.
- corresponding negative, positive, and additional viable tissues (without MTT addition) were treated identically without piercing.
- extract solution was mixed and two 200 μL aliquots were transferred into the appropriate wells of a 96-well plate(s).
- absorbance at 570 nm (OD570) of each well was measured with a plate reader (Versamax® Molecular Devices). No reference wavelength measurement was used.

DATA EVALUTION
1) mean optical density (OD) value of the blank control wells (ODBlk) for each experiment was calculated.
2) mean ODBlk from each mean OD value of the same experiment was subtracted (blank corrected values).
3) mean value of the two replicates for each tissue was calculated.
4) mean value of the two relating tissues for each control (negative control (NC) and positive control (PC) and test item (TI) was calculated (ODTI, ODNC, ODPC).
5) mean OD value of the negative control corresponds to 100% viability.
Corrected negative control OD = Negative Control OD - ODBlk = 100% Viability

6) percent viability of each of the two relating tissues for each control and test item relative to the negative control (= 100%) was calculated.
Viability (%) = 100 x ((ODTi/ODPC/ODNC) / mean ODNC)
7) difference of the viability between duplicate tissues was calculated (difference >20% the test is considered as non-valid).
8) mean viability of the test item (TI viability) was calculated and the test item was classified according to the prediction model.

PREDICTION MODEL
- test item-treated tissue viability is > 60% relative to the negative control treated tissue viability. Test item is identified as not requiring classification and labelling according to
GHS (No Category), i.e. is not an eye irritant.
- test item-treated tissue viability is ≤ 60% relative to negative control treated tissue viability. Test item is identified as potentially requiring classification and labelling according to UN GHS (Category 2 or Category 1), is an eye irritant.

ACCEPTABILITY CRITERIA
The results are acceptable, if:
1) negative control optical density is > 0.8 and < 2.5,
2) mean relative viability of the positive control is below 50% of the negative control viability.
3) difference of viability between the two relating tissues of a single test item is < 20% in the same run (for positive and negative control tissues and tissues of test items). This
applies also to the viabilities of freeze-killed tissues (items and negative control) and the additional viable tissues (without MTT addition) which are calculated relative to the negative control.
4) results of the positive and negative controls of the test method demonstrate reproducibility over time.

Irritation parameter:

other: % tissue viability

Value:

87.5

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

DEMONSTRATION OF TECHNICAL PROFICIENCY:
The laboratory had demonstrated technical proficiency by correctly predicting the irritation potential of fifteen proficiency chemicals listed in Table 1 of OECD TG 492.
Please also refer to the field "Attached background material" below.

OTHER EFFECTS:
- Colour interference with MTT: the colour interference pre-experiment to investigate the colour change potential of the test item in water or isopropanol did not result in a change in colour. Therefore, an additional test with viable tissues without MTT addition was not necessary.
- Direct-MTT reduction: optical evaluation of the MTT-reducing capacity of the test item with the MTT-reagent did not recognise a blue colour. Therefore, an additional test with freeze-killed tissues was not necessary.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes, negative control OD is > 0.8 and < 2.5 (values between 1.038 and 1.117).
- Acceptance criteria met for positive control: yes, mean relative viability of the positive control is below 50% of the negative control viability (37.0%).
- Acceptance criteria met for variability: yes, difference of viability between the two relating tissues of a single item is < 20% (values between 3.3% and 7.3%) in the same run (for positive and negative control tissues and tissues of single test items).
- Reproducibility of negative and positive control: results of the positive and negative controls of the test method demonstrate reproducibility over time. Even though the negative control absorbance is slightly below the historical range, it is in the acceptance range of OECD TG 492.

Table 1: Absorbance of tissues after treatment for 6 hours with the test item and the controls

Dose Group	Absorbance Well 1 (Tissue 1/2)	Absorbance Well 2 (Tissue 1/2)	Mean Absorbance* (Tissue 1/2)	Mean Absorbance* Tissue 1 and 2 minus Mean Blank	Mean Absorbance of 2 Tissues*	Rel. Ab- sorbance [%] Tissue 1 and 2**	Absolute Value of the Difference of the Rel. Absorbances [%] Tissue 1 and 2	Mean Rel. Absorbance [% of Negative Control]**
Blank	0.039	0.039	0.039	0.000
Negative Control	1.079	1.038	1.059	1.020	1.045	97.5	4.9	100.0
	1.103	1.117	1.110	1.071		102.5
Positive Control	0.440	0.446	0.443	0.404	0.387	38.6	3.3	37.0
	0.404	0.412	0.408	0.369		35.3
Test Item	0.950	1.033	0.992	0.953	0.914	91.1	7.3	87.5
	0.915	0.915	0.915	0.876		83.8

* Mean of two replicate wells after blank correction

** Relative absorbance [rounded values]: (100 X (absorbance test item/positive control))/ (absorbance negative control)

Table 2: Historical control data

Positive Control		Negative Control [OD570]
Mean Viability	32.47%	Mean Absorption	1.49
Rel. Standard Deviation	10.29%	Rel. Standard Deviation	0.24
Range of Viabilities	15.90% - 42.30%	Range of Absorbance	1.24 – 2.05
Mean Absorption	0.48
Rel. Standard Deviation	0.14
Range of Absorbance	0.22- 0.64

Data of 15 studies performed from July 2015 until end of December 2016

Interpretation of results:

GHS criteria not met

Conclusions:

Aluminium lanthanum trioxide does not possess any eye irritating potential. Aluminium lanthanum trioxide is identified as not requiring classification and labelling for eye irritation according to UN GHS (No Catogory).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Justification for classification or non-classification

Skin and eye irritation:

Aluminium lanthanum trioxide does not possess an irritation potential and does not require classification as skin or eye irritant according to Regulation (EC) 1272/2008 and subsequent adaptations.