Reaction product of D-Glucopyranoside, methyl; esterified with octadecanoic acid, methyl ester

Endpoint summary

Description of key information

Skin corrosion in vivo: The registered substance was determined to be non-corrosive to rabbit skin (equivalent or similar to OECD 404 and EU Method B.4).

 

Skin irritation in vivo: The registered substance was determined to be non-irritant to rabbit skin (equivalent or similar to OECD 404 and EU Method B.4).

 

Eye irritation in vivo: The registered substance was determined to be non-irritant to the rabbit eye (equivalent or similar to OECD 405 and EU Method B.5).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 April 2015 to 24 April 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

Remarks:

Hsdlf:NZW

Details on test animals or test system and environmental conditions:

ANIMAL INFORMATION
- Two New Zealand White (Hsdlf:NZW) were supplied by Harlan Laboratories UK Ltd, Leicestershire, UK.
- At the start of the study the animals weighed 2.75 or 3.00 kg and were 12 to 20 weeks old.
- After an acclimatisation period of at least 5 days each animal was given a number unique within the study. The number was written with black indelible marker pen on the inner surface of the ear and on the cage label.

ANIMAL CARE AND HUSBANDRY
- The animals were individually housed in suspended cages.
- Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd, Oxon, UK) was allowed throughout the study.
- The diet and drinking water were not considered to contain any contaminant of a level that might have affected the purpose or integrity of the study.
- Temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70 % respectively.
- Lighting was controlled by a timer switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.
- The rate of air exchange was at least 15 changes per hour.
- Animals were provided with environmental enrichment items which were not considered 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:

clipped

Vehicle:

unchanged (no vehicle)

Remarks:

absorption of the test item was not determined

Controls:

not required

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

Four hours

Observation period:

72 hours

Number of animals:

Two

Details on study design:

MEASUREMENT OF pH
- The pH of the test item was determined prior to commencement of the study and the findings are shown in the table below.

TEST SYSTEM
- On the day before the test, two rabbits were clipped free of fur on the dorsal/flank area using veterinary clippers.
- Only animals where gross observation showed healthy intact epidermis were selected for the study.
- On the day of the investigation a suitable test site was selected on the back of each rabbit.
- Test item (0.5 mL) was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch.
- The patch was secured in position with a strip of surgical adhesive tape.
- To prevent animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset.
- Animals were returned to their cages for the duration of the exposure period.
- Four hours after application the corset and patches were removed from each animal and any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water.
- Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for primary irritancy and scored according to the scale shown in the table below.
- Any other skin reactions and clinical signs of toxicity, if present, were also recorded.
- Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

INTERPRETATION OF RESULTS
- The scores for erythema and edema at the 24 and 72-Hour readings were totaled for the two test
rabbits (8 values) and this total was divided by 4 to give the primary irritation index of the test
item. The test item was graded according to the following scheme devised by Draize, J.H.
(1959): Non-irritant (primary irritation index 0); mild irritant (primary irritation index > 0 to 2); moderate irritant (primary irritation index > 2 to 5); severe irritant (primary irritation index > 5 to 8).
- 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 considered to be corrosive to rabbit skin. Grading according to Draize may, therefore, not be applicable.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75038 male

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75039 male

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75038 male

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75039 male

Irritant / corrosive response data:

SKIN REACTIONS
- The individual scores for erythema/eschar and edema are given in Table 1 (attached).
- Very slight erythema and very slight edema were noted at both treated skin sites 1 hour after patch removal.
- The treated skin sites appeared normal at the 24-Hour observation.

Other effects:

BODY WEIGHT
- Individual body weights and body weight change are given in Table 2 (attached).
- Both animals showed expected gain in body weight during the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item produced a primary irritation index of 0.0 and was considered to be non-irritant to rabbit skin according to the Draize scheme. No corrosive effects were noted.

Executive summary:

GUIDELINE

The study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit in compliance with the OECD Guideline for the Testing of Chemicals No 404 “Acute Dermal Irritation/Corrosion” (adopted 24 April 2002) and Method B.4 Acute Toxicity (Skin Irritation) of Commission Regulation (EC) No 440/2008.

 

METHODS

Two animals were clipped free of fur on the dorsal/flank area using veterinary clippers and 0.5 mL of test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in place with a strip of surgical adhesive tape and the trunk of each rabbit was wrapped in an elasticated corset. Four hours after application, the corset and patches were removed from each animal and any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water. Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for primary irritancy and scored.

 

RESULTS

Very slight erythema and very slight oedema were noted at both treated skin sites one hour after patch removal. The treated skin sites appeared normal at the 24-hour observation.

 

CONCLUSION

The test item produced a primary irritation index of 0.0 and was considered to be non-irritant to rabbit skin according to the Draize scheme. No corrosive effects were noted.

Reference 4

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Justification for type of information:

See read-across justification attached in Section 13.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

erythema score

Remarks:

key study

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75038 male

Irritation parameter:

erythema score

Remarks:

key study

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75039 male

Irritation parameter:

edema score

Remarks:

key study

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 45038 male

Irritation parameter:

edema score

Remarks:

key study

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75039 male

Irritation parameter:

erythema score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24 h

Score:

1.5

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Irritation parameter:

erythema score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

72 h

Score:

1

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Irritation parameter:

edema score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24 h

Score:

0.8

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Irritation parameter:

edema score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

72 h

Score:

0.5

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Reference 5

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

December 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

not applicable

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

not applicable

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

ANIMALS
- Six New Zealand white rabbits, weighing approximately 2 kilograms and about 3 months of age, sex unspecified, were obtained from a suitably licensed dealer.
- Animals were checked carefully upon receipt for diarrhoea and dehydration, respiratory difficulties, postural deficiencies, and general condition.
- Animals were acclimated at least 4 days prior to test initiation.
- The animals were housed in galvanised or stainless-steel cages, in a temperature controlled room with a 12-hour light/dark cycle.
- Diet consisted of a growth and maintenance ration from a commercial producer, as well as water, ad libitum.
- Animals were identified through individual markings on the outer ear of each animal, as well as a cage label.

Type of coverage:

occlusive

Preparation of test site:

clipped

Remarks:

abraded and non-abraded

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

24 hours

Observation period:

72 hours

Number of animals:

Six

Details on study design:

STUDY DESIGN
- Twenty-four hours prior to test initiation, the animals were re- examined. Any animals in poor condition, and particularly animals with skin eruptions or dermal lesions, were not used.
- Animals were prepared for testing by close-clipping the skin of the mid-dorsal area of the trunk, between the scapulae and the pelvis, using a small animal clipper equipped with a #40 (surgical) head.
- Immediately prior to test initiation, the animals were placed in wooden restrainers.
- Two test sites, each 2.5 cm2, were chosen on opposite sides of the vertebral column.
- The test site on the left side of the animal remained intact; the test site on the right was further prepared by abrading with a sterile 22-gauge hypodermic needle. The abrasions were longitudinal epidermal incisions, sufficiently deep to penetrate the stratum corneum, but not so deep as to destroy the integrity of the derma, i.e., to cause bleeding.
- A single application (0.5 mL) of the test article was made to each test site.
- The test article was then covered with a 2.5cm surgical gauze pad, and the latter held in place with adhesive tape.
- After both test sites were treated, the entire trunk of each animal was encased in an impermeable occlusive wrapping fixed in place with adhesive tape. This aided in maintaining the test article and patches in position and prevented the evaporation of possible volatile components of the test article.
- The wrapping and test article were removed 24 hours following application. Remaining test article was gently wiped from the skin, and each test site was individually examined and scored at 24 and 72 hours for erythema and edema using the Draize skin scoring scale. (see Table 1, attached). The presence of effects not listed in the scoring scale was also noted.
- Following the 72-hour reading, the mean scores for 24 and 72-hour gradings were averaged to determine the primary skin irritation index. A Score of 5.0 or more indicates a primary dermal irritant (see Table 2, attached).

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24 h

Score:

1.5

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

72 h

Score:

1

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24 h

Score:

0.8

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

72 h

Score:

0.5

Max. score:

4

Reversibility:

other: not reported but observed effects scored at levels below that causing classification

Remarks on result:

other: non-abraded skin

Irritant / corrosive response data:

- Individual scores are given in Table 3 (attached).

Interpretation of results:

study cannot be used for classification

Conclusions:

This test article was not found to be a primary dermal irritant to rabbits under conditions of the test.

Executive summary:

METHODS

Six New Zealand white rabbits each received a single dermal application (0.5 mL) of a preparation containing 20 % of an analogue test item on two test sites; one abraded and one intact. The test sites were occluded for 24 hours and were observed individually for erythema, edema, and other effects 24 and 72 hours after application. Mean scores from the 24 and 72-hour reading were averaged to determine the primary irritation index. The test article was used as a 20 % gravimetric mineral oil suspension.

 

RESULTS

The test article was not found to be a primary dermal irritant to rabbits under conditions of the test.

.

Reference 6

Endpoint:

skin irritation / corrosion, other

Remarks:

skin corrosion in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

not applicable

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

not applicable

GLP compliance:

no

Species:

rabbit

Strain:

other: New Zealand

Details on test animals or test system and environmental conditions:

ANIMALS
- A group of six albino New Zealand rabbits (three males and three females) weighing 1.8 to 2.4 kg were used in the study.

Type of coverage:

occlusive

Preparation of test site:

clipped

Remarks:

intact skin

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

48 hours

Number of animals:

Six

Details on study design:

METHOD
- Test material (0.5 mL) was applied to clipped areas of intact skin.
- Applications were made under occlusive patches (1” x 1” gauze, covered by adhesive tape).
- Following application of the test material, the entire trunk of each animal was covered with an impermeable occlusive wrapping and animals were immobilised.
- The wrapping and test material were removed 4 hours following application.
- At 4 and 48 hours, the sites were individually examined and scored separately for erythema and edema (see Table 1, attached).
- Mean scores for the 4 and 48-hour gradings were averaged to determine final irritation indices (see Table 1A, attached).
- Corrosiveness seen at 4 and/or 48 hours alone indicates a corrosive material. Tissue destruction (corrosiveness) does not include merely sloughing of the epidermis or erythema, oedema or fissuring.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

other: 4/48 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

edema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

other: 4/48 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

- Individual scores are given in Table 3 (attached).

Other effects:

None reported

Interpretation of results:

GHS criteria not met

Conclusions:

The primary irritation index was reported as 0.0 and the 100% concentration test item was considered to be non-corrosive to skin.

Executive summary:

METHODS

Three male and three female New Zealand white rabbits each received a single dermal application (0.5 mL) of test material to intact skin. The test sites were occluded for 4 hours and were observed individually for erythema, oedema, and other effects at 4 and 48 hours after application. Mean scores from the 4 and 48-hour reading were averaged to determine the primary irritation index.

 

RESULTS

The primary irritation index was reported as 0.0 and the 100% concentration test item was considered to be non-corrosive to skin.

Reference 7

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

not applicable

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

not applicable

GLP compliance:

no

Species:

rabbit

Strain:

other: New Zealand

Details on test animals or test system and environmental conditions:

ANIMALS
- A group of six albino New Zealand rabbits (three males and three females) weighing 1.8 to 2.4 kg were used in the study.

Type of coverage:

occlusive

Preparation of test site:

clipped

Remarks:

intact and abraded skin

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

24 hours

Observation period:

72 hours

Number of animals:

Six

Details on study design:

METHOD
- Test material (0.5 mL) was applied to clipped areas of intact and abraded skin.
- Abrasions were longitudinal epidermal incisions that were sufficiently deep to penetrate the stratum corneum, but not so deep as to destroy the integrity of the derma.
- Applications were made under occlusive patches (1” x 1” gauze, covered by adhesive tape).
- Following application of the test material, the entire trunk of each animal was covered with an impermeable occlusive wrapping and animals were immobilised.
- The wrapping and test material were removed 24 hours following application.
- At 24 and 72 hours, the sites were individually examined and scored separately for erythema and edema (see Table 1, attached).
- Mean scores for 24 and 72-hour gradings were averaged to determine final irritation indices (see Table 1A, attached).

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h in 4/6 animals with abraded skin and 5/6 animals with non-abraded skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h in 6/6 animals

Remarks on result:

other: abraded and non-abraded skin

Irritant / corrosive response data:

- Individual scores are given in Table 2 (attached).

Other effects:

None reported

Interpretation of results:

study cannot be used for classification

Remarks:

observed effects scored at levels below that causing EU classification and reversibility in all animals with non-abraded skin can be assumed

Conclusions:

The primary irritation index was reported as 1.13 and the 100% concentration test item was considered to have potential for mild irritation.

Executive summary:

METHODS

Three male and three female New Zealand white rabbits each received a single dermal application (0.5 mL) of test item on two test sites; one abraded and one intact. The test sites were occluded for 24 hours and were observed individually for erythema, oedema, and other effects 24 and 72 hours after application. Mean scores from the 24 and 72-hour reading were averaged to determine the primary irritation index.

 

RESULTS

The primary irritation index was reported as 1.13 and the 100% concentration test item was considered to have potential for mild irritation.

.

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

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 April 2015 to 08 May 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

Remarks:

Hsdlf:NZW

Details on test animals or tissues and environmental conditions:

ANIMAL INFORMATION
- 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 2.09 or 2.85 kg and were 12 to 20 weeks old.
- After an acclimatisation period of at least 5 days each animal was given a number unique within the study which was written with black indelible marker pen on the inner surface of the ear and on the cage label.

ANIMAL CARE AND HUSBANDRY
- Animals were individually housed in suspended cages.
- Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study.
- Diet and drinking water were not considered to contain any contaminant of a level that might have affected the purpose or integrity of the study.
- Temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70 % respectively.
- Rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.
- Animals were provided with environmental enrichment items that were not considered to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

Single application

Observation period (in vivo):

72 hours

Details on study design:

MEASUREMENT OF pH
- The pH of the test item was determined prior to commencement of the study and the findings are shown in the table below.

PROCEDURE
- 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 opthalmoscope.
- Only animals free of ocular damage were used.
- Initially, a single rabbit was treated.
- A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to the test item application to provide a therapeutic level of systemic analgesia.
- Five minutes prior to test item application, a pre-dose anaesthesia of ocular anaesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.
- Test item (0.1 mL) was placed into the conjunctiva! 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 (attached).
- Eight hours after test item application, a subcutaneous injection of post-dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.
- After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated.
- Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation of Draize (1977) given in Appendix 2 (attached).
- 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 body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

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) (see Appendix 3, attached) was employed to determine 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 determination of the eye irritancy potential of the test item.
- If evidence of irreversible ocular damage is noted, the test item will be considered to be corrosive to the eye.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75050 male

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75060 male

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: 75050 male

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: 45060 male

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 75050 male

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 75060 male

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: 75050 male

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: 45060 male

Irritant / corrosive response data:

OCULAR REACTIONS
- Individual and group mean scores for ocular irritation are given in Table 1 and Table 2 (attached).
- No corneal or iridial effects were noted during the study.
- Moderate conjunctiva! irritation was noted in both treated eyes 1 hour after treatment with minimal conjunctiva! irritation noted at the 24 and 48-hour observations.
- Both treated eyes appeared normal at the 72-hour observation.

Other effects:

BODY WEIGHT
- Individual body weights and body weight change are given in Table 3 (attached).
- Both animals showed expected gain in body weight during the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item produced a maximum group mean score of 8.0 and was considered to be a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system

Executive summary:

GUIDELINE

The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit in compliance with the OECD Guideline for the Testing of Chemicals No 405 “Acute Eye Irritation/Corrosion” (adopted 02 October 2012) and Method B.5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No 440/2008.

 

METHODS

The test involved a single application of the test item to the non-irrigated eye of two rabbits. Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre-dose anaesthesia of ocular anaesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye. A volume of 0.1 mL of the test item was placed into the conjunctiva! 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 a six-point scale. Eight hours after test item application, a subcutaneous injection of post-dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.

After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation of Draize (1977). 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 body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

 

RESULTS

A single application of the test item to the non-irrigated eye of two rabbits produced iridial inflammation and moderate conjunctival irritation. Both treated eyes appeared normal at the 72-hour observation.

 

CONCLUSION

The test item produced a maximum group mean score of 8.0 and was considered to be a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

Reference 3

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Justification for type of information:

See read-across justification attached in Section 13.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

cornea opacity score

Remarks:

key study

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75050 male

Irritation parameter:

cornea opacity score

Remarks:

key study

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 75060 male

Irritation parameter:

iris score

Remarks:

key study

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: 75050 male

Irritation parameter:

iris score

Remarks:

key study

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: 75060 male

Irritation parameter:

conjunctivae score

Remarks:

key study

Basis:

animal #1

Remarks:

conjunctival redness

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 75050 male

Irritation parameter:

conjunctivae score

Remarks:

key study

Basis:

animal #2

Remarks:

conjunctival redness

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 75060 male

Irritation parameter:

chemosis score

Remarks:

key study

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: 75050 male

Irritation parameter:

chemosis score

Remarks:

key study

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: 75060 male

Irritation parameter:

cornea opacity score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

iris score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Remarks on result:

other: conjunctival redness

Irritation parameter:

chemosis score

Remarks:

supporting study

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Reference 4

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

December 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not applicable

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

not applicable

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

ANIMALS
- Six New Zealand white rabbits, weighing approximately 2 kilograms and about 3 months of age, were obtained through a suitably licensed dealer.
- The animals were checked carefully upon receipt for ocular defects, diarrhoea and dehydration, respiratory difficulties, postural deficiencies, and general condition. Any animal exhibiting visible ocular defects or irritation, or poor condition, was not used in this test.
- Animals were acclimated for at least 3 days prior to test initiation. They were housed in galvanised or stainless-steel cages and identified through individual markings on the outer ear of each animal, as well as a cage label.
- Diet consisted of a growth and maintenance ration from a commercial producer, as well as water, ad libitum.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

Six

Details on study design:

STUDY DESIGN
- Immediately prior to test initiation, the animals were placed in wooden restrainers.
- Test article (0.1 mL) was placed in one eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test article was dropped. The eyelids were gently held together for one second.
- The contralateral eye, remaining untreated and served as a control.
- If any of the test article remained in the eye for 24 hours, the eye was washed out with lukewarm wafer after the 24-hour reading.
- Observations of ocular irritation were recorded 24, 48, and 72 hours following instillation of the test article (see Table 4, attached). Additional readings were made at 4 and 7 days after application if irritation persisted.
- An animal was considered as exhibiting a positive reaction if the test article produced any of the following: ulceration of the cornea other than a fine stippling, opacity of the cornea other than a slight dulling of the normal lustre, inflammation of the iris other than a slight deepening of the folds or slight circumcorneal injection of the blood vessels, obvious conjunctival swelling with partial eversion of the lids or a diffuse crimson-red with individual vessels not discernible.
- If two or more animals exhibited a positive reaction, the test article was considered an ocular irritant (unless the test was repeated with another six animals without positive reactions).

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

- See Tables 5 and 6 (attached).

Interpretation of results:

GHS criteria not met

Conclusions:

The test article was not found to be an ocular irritant to rabbits under conditions of the test.

Executive summary:

METHODS

Six New Zealand white rabbits, free from visible ocular defects, each received a single intraocular application (0.1 mL) of a preparation containing 20 % of an analogue test item in one eye. The contralateral eye, remaining untreated and served as a control. The eyes of all animals remained unwashed for 24 hours. Observations of corneal opacity, iritis, and conjunctivitis were recorded 24, 48, and 72 hours after treatment, and at 4 and 7 days if irritation persisted. The test article was used as a 20 % gravimetric mineral oil suspension.

 

RESULTS

The test article was not found to be an ocular irritant to rabbits under conditions of the test.

Reference 5

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not applicable

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

not applicable

GLP compliance:

no

Species:

rabbit

Strain:

other: New Zealand

Details on test animals or tissues and environmental conditions:

ANIMALS
- A group of six albino New Zealand rabbits, of mixed sex, weighing 1.8 to 2.4 kg were used in the study.

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1

Duration of treatment / exposure:

Single dose

Observation period (in vivo):

3 days

Number of animals or in vitro replicates:

Six

Details on study design:

METHOD
- Healthy New Zealand rabbits without ocular defects were used in the study.
- Test item (0.1 mL) was instilled into the right eye of each animal.
- The left eye of each animal remained untreated and served as a control.
- The treated eyes of all rabbits remained unwashed.
- Observations were made on the cornea, iris and the bulbar and palpebral conjunctivae.
- Numerical scores (see Table 4, attached) were assigned to lesions observed according to a standard scoring system in which injuries to the cornea and iris account for approximately 80 % of the total score. These structures are purposely weighted because of their vital role in vision.
- Readings facilitated by hand-held lenses were made 1, 2 and 3 days after treatment, and up to 7 days when necessary.

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

animals 1 to 6

Time point:

24/48/72 h

Score:

0

Max. score:

110

Reversibility:

other: not applicable

Irritant / corrosive response data:

- Individual scores are shown in Table 5 (attached)

Interpretation of results:

GHS criteria not met

Conclusions:

No evidence of eye damage/irritation was observed under the conditions of the test.

Executive summary:

METHODS

Six New Zealand white rabbits, free from visible ocular defects, each received a single intraocular application (0.1 mL) of test item in one eye. The contralateral eye, remaining untreated and served as a control. The eyes of all animals remained unwashed. Observations were made on the cornea, iris and the bulbar and palpebral conjunctivae at 1, 2 and 3 days after treatment. Further observations were made at 7 days after treatment if necessary.

RESULTS

No evidence of eye damage/irritation was observed under the conditions of the test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin corrosion in vitro

An in vitro skin corrosion study does not need to be conducted because adequate data are available from investigation of skin corrosion in vivo.

 

Skin corrosion in vivo

The key study was performed to assess the skin corrosion potential of the registered substance using a method equivalent or similar to OECD 404 and EU Method B.4.

 

Three male and three female New Zealand white rabbits each received a single dermal application (0.5 mL) of test material to intact skin. The test sites were occluded for 4 hours and were observed individually for erythema, oedema, and other effects at 4 and 48 hours after application. Mean scores from the 4 and 48-hour reading were averaged to determine the primary irritation index.

 

The primary irritation index was reported as 0.0 and the 100% concentration test item was considered to be non-corrosive to skin.

 

Skin irritation in vitro

An in vitro skin irritation study does not need to be conducted because adequate data are available from in vivo skin irritation studies.

 

Skin irritation in vivo

 

The first key study was performed to assess the irritancy potential of the registered substance using a method equivalent or similar to OECD 404 and EU Method B.4, six New Zealand white rabbits each received a single dermal application (0.5 mL) of test item on two test sites; one abraded and one intact. The test sites were occluded for 24 hours and were observed individually for erythema, oedema, and other effects 24 and 72 hours after application. Mean scores from the 24 and 72-hour reading were averaged to determine the primary irritation index.

 

The primary irritation index was reported as 1.13 and the 100% concentration test item was considered to have potential for mild irritation. However, observed effects were scored at levels below that causing EU classification and reversibility in all animals with non-abraded skin can be assumed.

A second key study was performed to assess the irritancy potential of an analogue test item to the skin of the New Zealand White rabbit in compliance with the OECD Guideline for the Testing of Chemicals No 404 “Acute Dermal Irritation/Corrosion” (adopted 24 April 2002) and Method B.4 Acute Toxicity (Skin Irritation) of Commission Regulation (EC) No 440/2008.

Two animals were clipped free of fur on the dorsal/flank area using veterinary clippers and 0.5 mL of test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in place with a strip of surgical adhesive tape and the trunk of each rabbit was wrapped in an elasticated corset. Four hours after application, the corset and patches were removed from each animal and any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water. Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for primary irritancy and scored.

 

Very slight erythema and very slight oedema were noted at both treated skin sites one hour after patch removal. The treated skin sites appeared normal at the 24-hour observation. The test item produced a primary irritation index of 0.0 and was considered to be non-irritant to rabbit skin according to the Draize scheme. No corrosive effects were noted.

In a supporting study, Six New Zealand white rabbits each received a single dermal application (0.5 mL) of a preparation containing 20 % of the same analogue test item on two test sites; one abraded and one intact. The test sites were occluded for 24 hours and were observed individually for erythema, edema, and other effects 24 and 72 hours after application. Mean scores from the 24 and 72-hour reading were averaged to determine the primary irritation index. The test article was used as a 20 % gravimetric mineral oil suspension. The test article was not found to be a primary dermal irritant to rabbits under conditions of this test.

Eye damage/irritation in vitro

An in vitro study to assess eye damage/irritation does not need to be conducted because adequate data are available from in vivo eye irritation studies.

 

Eye damage/ irritation in vivo

The first key study was performed to assess the irritancy potential of the registered substance using a method equivalent or similar toOECD 405 and EU Method B.5.

 

Six New Zealand white rabbits, free from visible ocular defects, each received a single intraocular application (0.1 mL) of test item in one eye. The contralateral eye, remaining untreated and served as a control. The eyes of all animals remained unwashed. Observations were made on the cornea, iris and the bulbar and palpebral conjuntivae at 1, 2 and 3 days after treatment. Further observations were made at 7 days after treatment if necessary.

 

No evidence of eye damage/irritation was observed under the conditions of the test.

A second key study was performed to assess the irritancy potential of an analogue test item to the eye of the New Zealand White rabbit in compliance with the OECD Guideline for the Testing of Chemicals No 405 “Acute Eye Irritation/Corrosion” (adopted 02 October 2012) and Method B.5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No 440/2008.

The test involved a single application of the test item to the non-irrigated eye of two rabbits. Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre-dose anaesthesia of ocular anaesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye. A volume of 0.1 mL of the test item was placed into the conjunctiva! 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 a six-point scale. Eight hours after test item application, a subcutaneous injection of post-dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required. After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated. Assessment of ocular damage/irritation was made approximately 1, 24, 48 and 72 hours following treatment, according to the numerical evaluation of Draize (1977). 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 body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

A single application of the test item to the non-irrigated eye of two rabbits produced iridial inflammation and moderate conjunctival irritation. Both treated eyes appeared normal at the 72-hour observation. The test item produced a maximum group mean score of 8.0 and was considered to be a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

 

In a supporting study, six New Zealand white rabbits,free from visible ocular defects, each received a single intraocular application (0.1 mL) of a preparation containing 20 % of an analogue test item in one eye. The contralateral eye, remaining untreated, served as a control. The eyes of all animals remained unwashed for 24 hours. Observations of corneal opacity, iritis, and conjunctivitis were recorded24,48, and 72 hours after treatment, and at 4 and 7 days if irritation persisted. The test article was used as a 20 % gravimetric mineral oil suspension.The test article was not found to be an ocular irritant to rabbits under conditions of the test.

Justification for classification or non-classification

Skin corrosion in vivo

A key in vivo study involving the registered substance at a concentration of 100 % reported the primary irritation index as 0.0. Classification for skin corrosion is therefore not required under the terms of Regulation 1272/2008.

 

Skin irritation in vivo

Available human data provides no evidence of skin irritation caused by the registered substance and a key in vivo study involving the registered substance at a concentration of 100 % permitted reversibility of low-level erythema within a short time frame to be assumed. These data are supported by a second key in vivo study on an analogue substance where mean scores (24/48/72 hours) were zero for erythema and oedema. The non-irritant nature of theregistered substance is further supported by a non-guideline study involving application of the same analogue test item to non-abraded rabbit skin in a preparation containing 20 % active ingredient. Classification of the registered substance for skin irritation is therefore unnecessary under the terms of Regulation 1272/2008.

Eye damage/irritation in vivo

A key in vivo study involving the registered substance at a concentration of 100 % reported no evidence of ocular effects. These data are supported by a second key study in whichan analogue substance was investigated in vivo. Mean scores (24/48/72 hours) were zero for corneal opacity and iritis and < 2 for conjunctival redness and chemosis with reported effects fully reversing within 72 hours. The non-irritant nature of the registered substance is further supported by a non-guideline study where the same analogue test item was instilled into rabbit eyes as a preparation containing 20 % active ingredient. Classification of the registered substance for eye damage/irritation is therefore unnecessary under the terms of Regulation 1272/2008.