Registration Dossier

Administrative data

Description of key information

As no data was available for eye or skin irritation of (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium, these endpoints were read across from aluminum, benzoate C16-18-fatty acids complexes. The read across in vitro eye irritation study on the enucleated rabbit eye indicated that (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium is unlikely to be irritant or corrosive to the eye. The read across in vivo study showed the material will be mildly irritant to rabbit eyes but below the threshold for classification.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
28 August 2101 to 03 September 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-compliant, guideline study, available as an unpublished report.
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
other: EPISKIN (TM) reconstructed human epidermis model
Strain:
other: Not applicable
Details on test animals and environmental conditions:
Not applicable
Type of coverage:
other: Topical
Preparation of test site:
other: Not applicable
Vehicle:
unchanged (no vehicle)
Controls:
other: Not applicable
Amount / concentration applied:
- Treatment group: Test carried out in triplicate. Approximately 10 mg of the test item was applied topically, ensuring an even covering, to the epidermis surface which had previously been moistened with 5 µL sterile, distilled water to improve contact between the solid test item and the epidermis.
- Negative control: 10 µL Dulbecco's Phosphate Buffered Saline (DPBS) with Ca++ and Mg++
- Positive control: 10 µL Sodium Dodecyl Sulphate (SDS) at 5% w/v aqueous solution spread over entire surface of the epidermis using a pipette tip with the process being repeated after 7 minutes.
Duration of treatment / exposure:
- Treatment period: 15 minutes
- Post-exposure incubation: At the end of the exposure period, tissues were rinsed with DPBS with Ca++ and Mg++. Rinsing was achieved by filling and emptying each tissue insert for approximately 40 seconds using a soft stream of DPBS to gently remove any residual test item. The rinsed tissues were transferred to a second column of three wells, each containing 2 mL of maintenance medium, and incubated for 42 hours, at 37°C and 5% CO2 in air.
Observation period:
Not applicable
Number of animals:
Not applicable
Details on study design:
Triplicate samples of epidermis tissue, previously moistened with 5µL sterile distilled water to improve contact with the solid test item, were uniformly covered with approximately 10 mg of test item. At the end of the 15 minute exposure period, each tissue was removed and rinsed with DPBS with Ca++ and Mg++ . The tissues were then incubated for 42 hours at 37°C and 5% CO2 in air, in wells containing 2 mL maintenance media.

The measurement of tissue viability (cytotoxicity) was measured by means of the colourimetric MTT reduction assay. Cell viability was measured by enzymatic reduction of the yellow MTT tetrazolium salt to a blue formazan salt (by the mitochondrial succinate dehdroganase in viable cells) in the test item treated tissue relative to the negative control using the following procedure. After incubation, the triplicate tissue samples were transferred into three wells each containing 2ml of a 0.3 mg/L MTT solution, care being taken to remove any excess maintenance medium from the bottom of the tissue insert by blotting on absorbent paper. The tissues were incubated for 3 hours at 37ºC, 5% CO2 in air and then the epidermis was carefully separated from the collagen matrix using forceps and both parts placed into 1.5 mL microtubes containing 500 µL of acidified isopropanol. Each tube was plugged and mixed thoroughly on a vortex mixer. The tubes were refrigerated at 1 to 10ºC for 3 days to allow the extraction of formazan crystals out of the MTT-loaded tissues. At the end of the formazan extraction period each tube was thoroughly mixed on a vortex mixer to produce a homogeneous coloured solution. The optical density of the extracted solution was measured at 540 nm against an acidified isopropanol blank.

The relative mean viability were calculated using the following equation;

Relative mean viability (%) = (mean OD540 of test item/mean OD540 of negative control) x 100.

If the mean tissue viability is ≤50%, the substance is considered to be irritant.
Irritation / corrosion parameter:
other:
Value:
98.4
Remarks on result:
other:
Remarks:
Basis: mean. Time point: After a 15-minute exposure. Max. score: 101.1. Reversibility: no data. Remarks: Negative control item (set at 100). Standard deviation relative to mean viability ± 3.4.. (migrated information)
Irritant / corrosive response data:
- Viability: The relative mean (n = 3) viability of the treated tissues was 98.4 after a 15-minute exposure period, with standard deviation of 3,4%
- Optical density: The mean optical density of the treated tissues at 540 nm was 0.777
- Conclusion: Test item is considered to be non-irritant using the EPISKIN (TM) human epidermis model (viability >50%)
Other effects:
- Positive control: The relative mean tissue viability was 11.8 % relative to the negative control and the standard deviation was 3.8%
- Negative control: The mean optical density was 0.790 and the standard deviation was 0.046.

-Direct MTT reduction: The MTT solution containing the test item did not turn blue, indicating that test item did not directly reduce MTT.

The acceptance criteria were satisfied according to the protocol criteria for both the positive control (relative mean viability ≤ 40% and standard deviation ≤ 18%) and negative control (OD540≥ 0.6 and standard deviation of individual tissue viability ≤18%). The standard deviation of the triplicate treated tissues was 3.4 % and hence the test item acceptance criterion (≤ 18%) was also satisfied.

Table 1. Mean OD540Values and Percentage Viability for Negative Control Item, Positive Control Item and Test Item

Item

OD540of tissue

Mean OD540of triplicate tissues

±SD of OD540

Relative Individual tissue viability (%)

Relative mean viability (%)

±SD of Relative mean viability (%)

Negative Control Item1

0.843

0.790

0.046

106.7

100*

5.5

0.758

97.2

0.760

96.2

Positive Control Item1

0.119

0.093

0.030

15.1

11.8

3.8

0.100

12.7

0.060

7.6

Test Item

0.785

0.777

0.027

99.4

98.4

3.4

0.747

94.8

0.799

101.1

SD = Standard deviation

* = The mean viability of the negative control tissues is set at 100%

1Control group shared with test laboratory project number 41202604

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Aluminum, benzoate C16-18-fatty acids complexes is considered to be non-irritant using the EPISKIN (TM) human epidermis model.
Executive summary:

Aluminum, benzoate C16-18-fatty acids complexesis considered to be non-irritant using the EPISKIN (TM) human epidermis model. The skin irritation potential of the test item was evaluated using EPISKIN (TM) reconstructed human epidermis model after a treatment period of 15 minutes followed by a post-exposure period of 42 hours using a colourimetric MTT reduction assay following OECD guideline 439 in an experimental proprietary study (Harlan 2013). The quality criteria required for the acceptance of results in the test were satisfied and the study is considered reliable and relevant for use.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
12 November 2012 to 22 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant, guideline study, available as an unpublished report.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Two New Zealand White rabbits were supplied by Harlan Laboratories UK Limited, Hillcrest, Belton, Loughborough, UK. At the start of the study the animals weighed 2.22 or 2.83 kg and were twelve to twenty weeks old. After an acclimatisation period of at least five days each animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.
The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 17 to 23°C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
A volume of 0.1 ml of the test material, which was found to weigh approximately 62 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye.
Duration of treatment / exposure:
Up to 72 hours
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
2
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: Not applicable

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 described by Kay J H and Calandra J C (1962), J. Soc. Cosmet. Chem. 13, 281 289 (see Appendix 3) 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.
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Highest score observed at 1 and 24 hour timepoints
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 24 hours
Remarks on result:
other: Highest score observed at 1 hour timepoint
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Highest score observed at 1 and 24 hour timepoints
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Highest score observed at 1 hour timepoint
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effect noted
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effect noted
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
72665 Male
Time point:
other: mean at 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: No effect noted
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: No effect noted
Irritation parameter:
other: Conjunctivae Discharge
Basis:
animal #1
Remarks:
72655 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
3
Reversibility:
other: No effects noted
Irritation parameter:
other: Conjunctivae Discharge
Basis:
animal #1
Remarks:
72703 Male
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
3
Reversibility:
fully reversible within: 1 hour
Remarks on result:
other: Highest score observed at 1 hour timepoint
Irritant / corrosive response data:
Individual and group mean scores for ocular irritation are given in Table 1 and Table 2 in any other information on results section. Individual mean scores for cornea, iris and conjunctivae are given in Table 3.
No corneal or iridial effects were noted during the study.
Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye one hour after treatment with minimal conjunctival irritation noted in both treated eyes at the 24 Hour observation. Both treated eyes appeared normal at the 48 Hour observation.
Other effects:
Both animals showed expected gain in bodyweight during the study (Table 4).

Table 1              Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

72665 Male

72703 Male

IPR= 2

IPR = 1

Time After Treatment

1
Hour

24
Hours

48
Hours

72
Hours

1
Hour

24
Hours

48
Hours

72
Hours

CORNEA

 

 

 

 

 

 

 

 

E = Degree of Opacity

0

0

0

0

0

0

0

0

F = Area of Cornea Involved

0

0

0

0

0

0

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

IRIS

 

 

 

 

 

 

 

 

D

0

0

0

0

0

0

0

0

Score (D x 5)

0

0

0

0

0

0

0

0

CONJUNCTIVAE

 

 

 

 

 

 

 

 

A = Redness

1

1

0

0

2

1

0

0

B = Chemosis

1

1

0

0

2

1

0

0

C = Discharge

0

0

0

0

1

0

0

0

Score (A + B + C) x 2

4

4

0

0

10

4

0

0

Total Score

4

4

0

0

10

4

0

0

IPR=  Initial pain reaction

Interpretation According to the Globally Harmonized System of Classification and Labelling of Chemical

Irreversible eye effects categories

An eye irritant Category 1 (irreversible effects on the eye)is a test material that produces:

-

at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days; and/or

-

at least in 2 of 3 tested animals a positive response of:

corneal opacity³ 3 and/or

iritis³ 1.5r

calculated as the mean scores following grading at 24, 48 and 72 hours after instillation of the test material

Reversible eye effects categories

An eye irritant Category 2A (irritating to eyes)is a test material that produces:

-

at least in 2 of 3 tested animals a positive response of:

corneal opacity³ 1 and/or

iritis³ 1 and/or

conjunctival redness³ 2 and/or

conjunctival oedema (chemosis)³ 2

-

Calculated as the mean scores following grading at 24, 48 and 72-Hour observations after instillation of the test material, and

at

-

which fully reverses within an observation period of normally 21 days

Within this category an eye irritant isconsidered mildlyirritating to eyes (Category 2B)when the effects listed above are fully reversible within 7 days of observation.

Interpretation According to Regulation (EC) No 1272/2008, Relating to the Classification, Labelling and Packaging of Dangerous Substances

        

Category for irreversible eye effects

Category

Criteria

Irreversible effects on the eye

(Category 1)

If, when applied to the eye of an animal, a substance produces:

-  at least in one animal effects on the cornea, iris or conjunctiva that   are not expected to reverse or have not fully reversedwithin an observation periodof normally 21 days;

and/or

-     at least in 2 of 3 animals, a positive response of:

-     cornea opacity³3 and/or

-     iritis>1.5

calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.

Category for reversible eye effects

Category

Criteria

Irritating to eyes

(Category 2)

If, when applied to the eye of an animal, a substance produces:

-     at least in 2 of 3 tested animals, a positive response of:

-   cornea opacity³1 and/or

-   iritis>1, and/or

-   conjunctival redness³2 and/or

-   conjunctival oedema (chemosis)³2

-   calculated as the mean scores following grading at 24, 48 and         72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

For those substances where there is pronounced variability among animal responses, this information shall be taken into account in determining the classification.

Table 2              Individual Total Scores and Group MeanScores forOcular Irritation

Rabbit Number

and Sex

Individual Total Scores At:

1 Hour

24 Hours

48 Hours

72 Hours

72665 Male

4

4

0

0

72703Male

10

4

0

0

Group Total

14

8

0

0

Group Mean Score

7.0

4.0

0.0

0.0

Table 3            Individual and Mean Scores for Cornea, Iris and Conjunctivae

Rabbit Number and Sex

Time After Treatment

Corneal Opacity

Iridial Inflammation

Conjunctival Redness

Conjunctival Chemosis

72665 Male

24 Hours

0

0

1

1

48 Hours

0

0

0

0

72 Hours

0

0

0

0

Total

0

0

1

1

Mean

0.0

0.0

0.3

0.3

72703 Male

24 Hours

0

0

1

1

48 Hours

0

0

0

0

72 Hours

0

0

0

0

Total

0

0

1

1

Mean

0.0

0.0

0.3

0.3

 

Table 4              Individual Bodyweights and Bodyweight Changes

Rabbit Number
and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 3

72665 Male

2.83

2.88

0.05

72703 Male

2.22

2.25

0.03

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other:
Conclusions:
The test material, aluminum, benzoate C16-18-fatty acids complexes, produced a maximum group mean score of 7.0 and was classified as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. Aluminum, benzoate C16-18-fatty acids complexes does not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals or according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.
Executive summary:

Introduction.

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The assessment was carried out in a GLP-compliant study following OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 24 April 2002) and Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008 in a proprietary, experimental study (Harlan 2013). The study is considered reliable and relevant for use for this endpoint.

Result.

A single application of the test material to the non-irrigated eye of two rabbits produced moderate conjunctival irritation in one treated eye and minimal conjunctival irritation in the other treated eye one hour after treatment with minimal conjunctival irritation noted in both treated eyes at the 24-hour observation. Both treated eyes appeared normal at the 48-hour observation.

Conclusion.

The test material, aluminum, benzoate C16-18-fatty acids complexes, produced a maximum group mean score of 7.0 and was classified as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. Aluminum, benzoate C16-18-fatty acids complexesdoes not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals or according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

As no data was available for eye or skin irritation of (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium, these endpoints were read across from aluminum, benzoate C16-18-fatty acids complexes.

Aluminum, benzoate C16-18-fatty acids complexes (CAS No. 94166-87-7, EC No. 303-385-6) is considered suitable for read-across to (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium as it is structurally similar being an approximate 2:1 mixture of (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium and (benzoato-O,O')hydroxy(hexadecanoato-O,O')aluminium. The only significant difference is the presence of about 30% of the aluminium complex containing the saturated C16-fatty acid, hexadecanoate, in place of the saturated C18-fatty acid, octadecanoate. This small difference in composition is expected to have no adverse influence on skin or eye irritation.

 

The skin irritation potential of the read across substance was evaluated using the in vitro EPISKIN (TM) reconstructed human epidermis model after a treatment period of 15 minutes followed by a post-exposure period of 42 hours using a colourimetric MTT reduction assay following OECD guideline 439 in an experimental proprietary study. The quality criteria required for the acceptance of results in the test were satisfied and the study is considered reliable and suitable for use. No evidence of corrosivity or classifiable skin irritation was observed.

The occular irritation potential of the read across substance was first assessed in a GLP-compliant, non-guideline in vitro study on enucleated rabbit eyes. The test item was applied to the cornea of three enucleated rabbit eyes maintained at 32°C in a superfusion chamber and maximal ocular irritation observations were recorded at 60, 120 and 240 minutes, for corneal opacity, fluorescein uptake and condition of the corneal epithelium. The study is considered reliable and suitable for use for this endpoint. No evidence of corrosion was observed.

An in vivo study was performed to assess the irritancy potential of the read across substance to the eye of the New Zealand White rabbit. The assessment was carried out in a GLP-compliant study following OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 24 April 2002) and Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008 in a proprietary, experimental study. The study is considered reliable and suitable for use for this endpoint. No evidence of classifiable eye irritation was observed.

The isolated form of the test substance was used for these studies.


Justification for selection of skin irritation / corrosion endpoint:
Only one study available based on read across from GLP compliant study for aluminum, benzoate C16-18-fatty acids complexes.

Justification for selection of eye irritation endpoint:
Read-across data from an in vivo GLP study in rabbits for aluminum, benzoate C16 -18 fatty acids complexes have been taken as representative of eye irritation assessment.

Justification for classification or non-classification

(Benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium is considered to be non-irritant using the EPISKIN (TM) human epidermis model.When assessed for eye irritation in vivo, the read-across substance aluminium, benzoate C16-18 fatty acid complexes exhibited minor irritation which did not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals or according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.