Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Skin:

A study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit. The test item produced a primary irritation index of 2.0 and was classified as a mild irritant to rabbit skin according to the Draize classification scheme. However, the test item did not meet the criteria for classification according to the EU Regulation on the classification, labeling and packaging of substances and mixtures.

Eye:

Although a previous in vivo study (Dean, 1976) concluded that Cadox TS-50 would not be considered an eye irritant, this study done in a pre-GLP era, lacked full experimental details and failed to fully establish the test item's identity / purity. Therefore, the study conclusions cannot be relied upon.A negative result was obtained from an in vitro study (BCOP; Warren, 2012).A negative study in vitro eye test does not fully guarantee a completely negative in vivo test results, but this in vitro study was considered as a good supportive study. In the key study, an in vivo GLP test conducted in rabbits (Sanders, 2012), the test item did not meet the criteria for classification accordingtoRegulation (EC) No. 1272/2008 on the classification, labelling and packaging of substances and mixtures. It is thereforeconsidered a non-irritant.


Justification for selection of skin irritation / corrosion endpoint:
Apparently well-conducted GLP study in vivo in accordance with OECD guideline.

Justification for selection of eye irritation endpoint:
Apparently well-conducted in vivo GLP study in accordance with OECD guideline.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 24 April 2012 and 10 May 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.35 or 3.19 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 Certified Rabbit diet supplied by Harlan Laboratories UK Ltd., 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.
Type of coverage:
semiocclusive
Preparation of test site:
other: clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
A quantity of 0.5 ml of the test item was applied directly to the skin
Duration of treatment / exposure:
4 hours
Observation period:
7 days
Number of animals:
2
Details on study design:
On the day before the test each rabbit was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.
One rabbit was initially treated. Three suitable sites were selected on the back of the rabbit. A quantity of 0.5 ml of the test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. Each patch was secured in position with a strip of surgical adhesive tape. To prevent the animal interfering with the patches, the trunk of the rabbit was wrapped in an elasticated corset for the duration of the exposure period.
One patch was removed at each of three time points: 3 minutes, 1 hour and 4 hours after application. Any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water.
After consideration of the skin reactions produced in the first animal, an additional animal was treated with 0.5 ml of test item. One patch was applied to the back of the rabbit and was allowed to remain in contact with the skin for a period of four hours.
Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the following scale:
EVALUATION OF SKIN REACTIONS
Erythema and Eschar Formation Value
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema 4
Oedema Formation Value
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimetre) 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4
Any other skin reactions and clinical signs of toxicity, if present, were also recorded.
An additional observation was made on Day 7 to assess the reversibility of skin reactions.
Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Irritation parameter:
other: Erythema/Eschar Formation
Basis:
animal: 71953 male
Time point:
other: Highest score at 0, 24, 48, 72 hours and 7 days
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
other: Erythema/Eschar Formation
Basis:
animal: 71986 Male
Time point:
other: Highest score at 0, 24, 48, 72 hours and 7 days
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
other: Oedema Formation
Basis:
animal: 71953 male
Time point:
other: Highest score at 0, 24, 48, 72 hours and 7 days
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
other: Oedema Formation
Basis:
animal: 71986 male
Time point:
other: Highest score at 0, 24, 48, 72 hours and 7 days
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritant / corrosive response data:
3-Minute Exposure Period
The individual scores for erythema/eschar and oedema are given in Table 1.
No evidence of skin irritation was noted during the study.

1-Hour Exposure Period
The individual scores for erythema/eschar and oedema are given in Table 1.
No evidence of skin irritation was noted during the study.

4-Hour Exposure Period
The individual scores for erythema/eschar and oedema are given in Table 2.
Well-defined erythema and very slight oedema were noted at both treated skin sites at the 24 Hour observation. Well-defined erythema and very slight oedema persisted at one treated skin site with very slight erythema noted at the other treated skin site at the 48 Hour observation. Very slight erythema was noted at both treated skin sites at the 72 Hour observation.
Both treated skin sites appeared normal at the 7 Day observation.
Other effects:
Bodyweight
Individual bodyweights and bodyweight changes are given in Table 3.
Both animals showed expected gain in bodyweight during the study.

Interpretation of Results

Calculation of Primary Irritation Index and Grading of Irritancy Potential Using the Draize Scheme

The scores for erythema and oedema at the 24 and 72‑Hour readings were totalled for the two test rabbits (8 values) and this total was divided by four to give the primary irritation index of the test item. The test item was classified according to the following scheme devised by Draize J H (1959) "Dermal Toxicity" In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Association of Food and Drug Officials of the,,, p.46‑59:

Primary Irritation Index

Classification of Irritancy

0

Non-irritant

> 0 to 2

Mild irritant

> 2 to 5

Moderate irritant

> 5 to 8

Severe irritant

If irreversible alteration of the dermal tissue is noted in any rabbit, as judged by the Study Director, which include ulceration and clear necrosis or signs of scar tissue, the test item is classified as corrosive to rabbit skin. Classification according to Draize may, therefore, not be applicable.

The results were also interpreted according to Regulation (EC) No. 1272/2008 on the classification, labelling and packaging of substances and mixtures.

Table 1              Individual Skin Reactions Following 3-Minute and 1-Hour Exposures

Skin Reaction

Observation Time
(following patch removal)

Individual Scores - Rabbit Number and Sex

71953Male

3-Minute Exposure

1-Hour Exposure

Erythema/Eschar Formation

Immediately

0

0

1 Hour

0

0

24 Hours

0

0

48 Hours

0

0

72 Hours

0

0

7 Days

0

0

Oedema Formation

Immediately

0

0

1 Hour

0

0

24 Hours

0

0

48 Hours

0

0

72 Hours

0

0

7 Days

0

0

Table 2              IndividualSkin ReactionsFollowing 4-Hour Exposure

Skin Reaction

Observation Time
(following patch removal)

Individual Scores – Rabbit Number and Sex

Total

71953Male

71986Male

Erythema/Eschar Formation

Immediately

0

0

(0 )

1 Hour

0

0

( 0 )

24 Hours

2

2

4

48 Hours

2

1

( 3 )

72 Hours

1

1

2

7 Days

0

0

( 0 )

Oedema Formation

Immediately

0

0

( 0 )

1 Hour

0

0

( 0 )

24 Hours

1

1

2

48 Hours

1

0

( 1 )

72 Hours

0

0

0

7 Days

0

0

( 0 )

Sum of 24 and 72‑Hour(S)    :          8

Primary Irritation Index (S/4)                  :          8/4 = 2.0

Classification                                       :          MILD IRRITANT

(   ) = Total values not used for calculation of primary irritation index

Table 3              Individual Bodyweights and Bodyweight Changes

Rabbit Number
and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 7

71953Male

3.19

3.37

0.18

71986Male

2.35

2.46

0.11


Interpretation of results:
other: MILD IRRITANT
Remarks:
Criteria used for interpretation of results: other: Draize classification scheme
Conclusions:
The test item produced a primary irritation index of 2.0 and was classified as a MILD IRRITANT to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.
The test item did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the classification, labelling and packaging of substances and mixtures.
Executive summary:

Introduction. The study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit. The method was designed to be compatible with the following:

OECD Guidelines for the Testing of Chemicals No. 404 "Acute Dermal Irritation/Corrosion" adopted 24 April 2002)

Method B4 Acute Toxicity (Skin Irritation) of Commission Regulation (EC) No. 440/2008

Results. 3-minute and 1-hour semi-occluded applications of the test item to the intact skin of one rabbit produced no evidence of skin irritation.

A single 4-hour, semi-occluded application of the test item to the intact skin of two rabbits produced well-defined erythema and very slight oedema. Both treated skin sites appeared normal at the 7-Day observation. No corrosive effects were noted.

Conclusion. The test item produced a primary irritation index of 2.0 and was classified as a mild irritant to rabbit skin according to the Draize classification scheme.

The test item did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the classification, labelling and packaging of substances and mixtures.

Endpoint:
skin irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 26 June 2012 and 01 July 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.43 or 2.49 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 Certified Rabbit diet supplied by Harlan Laboratories UK Ltd., 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:
other: The left eye remained untreated and was used for control purposes.
Amount / concentration applied:
A volume of 0.1 ml of the test item was placed into the conjunctival sac of the right eye
Duration of treatment / exposure:
1 hour
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
2
Details on study design:
Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.
Initially, a single rabbit was treated. A volume of 0.1 ml of the test item was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1.
After consideration of the ocular responses produced in the first treated animal, a second animal was treated.
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation given in Appendix 2, (from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).
Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Any clinical signs of toxicity, if present, were also recorded.
Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Irritation parameter:
cornea opacity score
Basis:
animal: 72174 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
cornea opacity score
Basis:
animal: 72184 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
iris score
Basis:
animal: 72174 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: No effects observed
Irritation parameter:
iris score
Basis:
animal: 72184 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: No effects observed
Irritation parameter:
other: redness
Basis:
animal: 72174 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
other: redness
Basis:
animal: 72184 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal: 72174 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
chemosis score
Basis:
animal: 72184 Male
Time point:
other: Highest score at 1, 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 24 hours
Irritant / corrosive response data:
Individual and group mean scores for ocular irritation are given in Table 1 and Table 2.
No corneal or iridial effects were noted during the study.
Minimal conjunctival irritation was noted in both treated eyes one and 24 hours after treatment.
Both treated eyes appeared normal at the 48-Hour observation.
Other effects:
Individual bodyweights and bodyweight changes are given in Table 3.
Both animals showed expected gain in bodyweight during the study.

Interpretation of Results

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 (seeAppendix3) was used to classify the ocular irritancy potential of the test item. 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 item.

If evidence of irreversible ocular damage is noted, the test item will be classified as corrosive to the eye.

The results were also interpreted according to Regulation (EC) No. 1272/2008 on the classification, labelling and packaging of substances and mixtures.

Table 1              Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

72174Male

72184Male

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

1

1

0

0

B = Chemosis

1

1

0

0

1

0

0

0

C = Discharge

1

0

0

0

0

0

0

0

Score (A + B + C) x 2

6

4

0

0

4

2

0

0

Total Score

6

4

0

0

4

2

0

0

IPR=  Initial pain reaction

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

Rabbit Number

and Sex

Individual Total Scores At:

1 Hour

24 Hours

48 Hours

72 Hours

72174Male

6

4

0

0

72184Male

4

2

0

0

Group Total

10

6

0

0

Group Mean Score

5.0

3.0

0.0

0.0

Table 3              Individual Bodyweights and Bodyweight Changes

Rabbit Number
and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 3

72174Male

2.43

2.55

0.12

72184Male

2.49

2.63

0.14

Interpretation of results:
other: minimal irritant (Class 3 on a 1 to 8 scale)
Remarks:
Criteria used for interpretation of results: other: modified Kay and Calandra classification system
Conclusions:
The test item produced a maximum group mean score of 5.0 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.
The test item did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the classification, labelling and packaging of substances and mixtures.
Executive summary:

Introduction. The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit. The method was designed to be compatible with the following:

- OECD Guideline for the Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion (adopted 24 April 2002)

- Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No.440/2008

Result. A single application of the test item to the non-irrigated eye of two rabbits produced minimal conjunctival irritation. Both treated eyes appeared normal at the 48‑Hour observation.

Conclusion. The test item produced a maximum group mean score of 5.0 and was classified as aminimal irritant (Class3 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. The test item did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the classification, labelling and packaging of substances and mixtures.

Endpoint:
eye irritation: in vitro / ex vivo
Remarks:
ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was performed on 22 March 2012.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.
Qualifier:
according to guideline
Guideline:
other: OECD 437
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
other: Excised Bovine Cornea
Strain:
other: Not Applicable
Details on test animals or tissues and environmental conditions:
Not applicable
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL

-Amounts(s) applied (volume or weight with unit):
0.75 mL of the test material was applied to triplicate corneas.

-Concentration (if solution):
The test material was used as supplied.

VEHICLE
No vehicle used
Duration of treatment / exposure:
The undiluted test material was applied for 10 minutes followed by an incubation period of 120 minutes.
Observation period (in vivo):
Not applicable
Number of animals or in vitro replicates:
Not applicable
Details on study design:
TEST SITE
-Area of exposure
0.75 mL of the test material was applied to triplicate corneas.

-% coverage:
The test material was topically applied to the cornea. The holders were gently tilted back and forth to ensure a uniform application of the material over the entire cornea.

-Type of wrap used:
None used

REMOVAL OF TEST SUBSTANCE
-Washing (if done):
At the end of the exposure period the test material was removed from the anterior chamber and each cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete MEM.

-Time after start of exposure:
10 minutes post exposure

SCORING SYSTEM:
Results from the two test method endpoints, opacity and permeability, were combined in an empirically derived formula to generate an In Vitro Irritancy Score.
Opacity Measurement-
The change in opacity for each cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final opacity reading. These values were then corrected by subtracting from each the average change in opacity observed for the negative control corneas. The mean opacity value of each treatment group was then calculated by averaging the corrected opacity values of each cornea for that treatment group.

Permeability Measurement
The corrected OD492 was calculated by subtracting the mean OD492 of the negative control corneas from the OD492 value of each treated cornea. The OD492 value of each treatment group was calculated by averaging the corrected OD492 values of the treated corneas for the treatment group.

The following formula was used to determine the in vitro score:
In Vitro Irritancy Score = mean opacity value + (15 x mean OD492 value)

DATA INTERPRETATION
A test material that induces an in vitro irritancy score > than or equal to 55.1 is defined as an ocular corrosive or severe irritant.

Additionally, the opacity and permeability values were evaluated independently to determine whether the test material induced a response through only one of the two endpoints.

Visual Observation
The condition of the cornea was visually assessed immediately after rinsing and at the final opacity measurement.
Irritation parameter:
cornea opacity score
Remarks:
Mean Score of Opacity & Permeability
Run / experiment:
120 minutes, Post Rinsing
Value:
0.7
Vehicle controls validity:
not applicable
Negative controls validity:
not applicable
Positive controls validity:
not applicable
Remarks on result:
other: The corneas treated with the test item and negative control item were clear post treatment and post incubation.

RESULTS

Corneal Opacity and Permeability Measurement

Individual and mean corneal opacity measurements and individual and mean corneal permeability measurements are given in Table 1.

Corneal Epithelium Condition

The condition of the cornea immediately after rinsing and at the final opacity measurement is given in Table 2.

In Vitro Irritancy Score

The results are summarised as follows:

Treatment

In Vitro Irritancy Score

Test Material

0.7

Negative Control

2.8

Positive Control

47.8

Criteria for an Acceptable Test

The positive control In Vitro irritancy Score was within the range of 30.9 to 67.7. The positive control acceptance criterion was therefore satisfied.

Table 1          Individual and Mean Corneal Opacity and Permeability Measurements

Treatment

Cornea Number

Opacity

Permeability (OD)

In vitroIrritancy Score

Pre-Treatment

Post-Treatment

Post-Incubation

Post-Incubation-Pre‑Treatment

Corrected Value

 

Corrected Value

Negative Control

1

3

3

6

3

 

0.032

 

 

2

4

4

5

1

 

0.029

 

 

3

4

3

7

3

 

0.029

 

 

 

 

 

 

2.3*

 

0.030+

 

2.8

Positive Control

4

2

18

20

18

15.7

1.935

1.905

 

5

3

21

23

20

17.7

2.030

2.000

 

6

2

22

25

23

20.7

2.090

2.060

 

 

 

 

 

 

18.0·

 

1.988·

47.8

Test Item

7

2

3

3

1

0.0

0.072

0.042

 

8

2

3

5

3

0.7

0.042

0.012

 

9

2

2

5

3

0.7

0.036

0.006

 

 

 

 

 

 

0.4·

 

0.020·

0.7


OD= Optical density   * = Mean of the post treatment-pre‑treatment values    += Mean permeability         ·= Mean corrected value


Table 2          Corneal Epithelium Condition

Treatment

Cornea Number

Observation

Post Treatment

Post Incubation

Negative Control

1

clear

clear

2

clear

clear

3

clear

clear

Positive Control

4

cloudy

cloudy

5

cloudy

cloudy

6

cloudy

cloudy

Test Item

7

clear

clear

8

clear

clear

9

clear

clear


Interpretation of results:
other: Non-Corrosive
Remarks:
Criteria used for interpretation of results: expert judgment
Conclusions:
The test item was considered not to be an ocular corrosive or severe irritant.
Executive summary:

Introduction. 

A study was performed to assess the ocular irritancy potential of the test item to the isolated bovine cornea. The method was designed to be compatible with the following:

OECD Guidelines for the Testing of Chemicals No. 437 (2009) “Bovine Corneal Opacity and Permeability Assay”

Method. 

The undiluted test item was applied for 10 minutes followed by an incubation period of 120 minutes. Negative and positive control items were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). 

Results.

 The in vitro Irritancy scores are summarised as follows:

Treatment

In Vitro Irritancy Score

Test Item

0.7

Negative Control

2.8

Positive Control

47.8

Conclusion. 

The test item was considered not to be an ocular corrosive or severe irritant.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

In vitro and in vivo GLP tests conducted in rabbits, the test item did not meet the criteria as a skin or eye irritant. Therefore, the data are conclusive but not sufficient for classification.