Reaction products of alkenyl dihydrofuran-2,5-dione and polyisoalkenyl dihydro-2,5-furandione with alkylene glycol

Endpoint summary

Description of key information

The skin irritancy potential of the test material was assessed inaccordance with OCED Guideline 404.  The test material, produced a primary irritation index of 0.72 in relation to erythema only, and which was fully reversible by the 7-day observation period.  Therefore, the test material can be considered to be not irritiating and does not meet the GHS criteria for classification.  
The in vitro eye irritancy potential of the test material was assessed in accordance with OECD Guideline 437.  The test item was considered not to be an ocular corrosive or severe irritant.  The result of this study has identified the test item as not causing serious eye damage. Furthermore, The eye irritancy potential of the test item was assessed in accordance with OECD Guideline 405.  The test item did not cause any damage to the rabbit eye. The individual mean scores for corneal opacity and iris light reflex were respectively 0.00 and 0.5 for all two animals. The individual mean scores for the conjunctivae were 2 for reddening and 1.33 for chemosis for all animals. Based on these results, the test item is considered to be irritating to rabbit eye.  As such, the test material does not meet the GHS criteria for classification.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 June to 4 July 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

On one occasion the temperature was outside the protocol limit of 23'C. This deviation was considered not to affect the purpose or integrity of the study.

Reason / purpose for cross-reference:

read-across: supporting information

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)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: Supplied by sponsor
- Date Recieved: 1 June 1995


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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

At the start of the study the animals weighed 2.45 to 2.67 kg and were twelve to sixteen weeks old. After a minimum acclimatisation period of 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 metal cages. Free access to mains drinking water and food (STANRAB SQC Rabbit Diet, Special Diets Services Ltd., Witham, Essex, UK) was allowed throughout the study.
The animal room was maintained at a temperature of 18 to 24' C and relative humidity of 45 to 59%. The rate of air exchange was approximately 15
changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness. On one occasion the
temperature was outside the protocol limit of 23'C. This deviation was considered not to affect the purpose or integrity of the study.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL/animal

Duration of treatment / exposure:

4 Hours

Observation period:

72 hours

Number of animals:

6

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.
On the day of the test a suitable test site was selected on the back of each rabbit. A quantity of 0.5 ml of the test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the shorn skin. The patch was secured in position with a strip of surgical adhesive tape
(BLENDERM: approximate size 2.5 cm x 4.0 cm). To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an
elasticated corset (TUBIGRIP) and the 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 material removed by gentle swabbing with cotton wool soaked in distilled water.
Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the following scale from Draize J.H., (1 977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington D.C. p.31.
SCORING SYSTEM:The skin reaction was assessed according to the numerical scoring system listed in the Commission Regulation (EC) No. 440/2008

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 Days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0.75

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

positive indication of irritation

Remarks:

Mild Irritation

Irritant / corrosive response data:

Very slight to well-defined erythema was noted at all treated skin sites at the 1-hour observation with very slight erythema at the 24-hour observation. Very slight erythema was noted at four treated skin sites at the 48-hour observation and three treated skin sites at the 72-hour observation.
Very slight oedema was noted at all treated skin sites at the 1-hour observation.
Desquamation, which was considered to be reversible so did not warrant continuation of the observation period, was noted at four treated skin sites at the 7-day observation.

Other effects:

None

Interpretation of results:

GHS criteria not met

Conclusions:

The skin irritancy potential of the test material was assessed inaccordance with OCED Guideline 404. The test material, produced a primary irritation index of 0.72 in relation to erythema only, and which was fully reversible by the 7-day observation period. Therefore, the test material can be considered to be not irritiating and does not meet the GHS criteria for classification.

Executive summary:

Guideline

A study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method followed the recommendations of the US Environmental Protection Agency (EPA) Guidelines, OECD Guidelines for Testing of Chemicals No. 404 "Acute Dermal Irritation/Corrosion" (adopted 17 July 1992) and Method 84 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 6 7/548/E EC) and Austrialian Toxicology Guidelines.

Method

A single 4-hour, semi-occluded application of the test material to the intact skin of six rabbits produced very slight to well-defined erythema and very slight oedema. Desquamation was noted on day 7 but was considered to be a reversible effect which did not warrant continuation of the observation period.

Results

The test material, produced a primary irritation index of 0.72 in relation to erythema only, and which was fully reversible by the 7-day observation period.

The substance is not classified for skin irritation in accordance with the CLP Regulation 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 to 29 April 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

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

Species:

cattle

Details on test animals or tissues and environmental conditions:

Eyes from adult cattle were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee and placed in Hanks’ Balanced Salt Solution (HBSS), supplemented with Penicillin and Streptomycin, and transported to the laboratory on ice packs. The corneas were refrigerated on arrival and used within 24 hours of receipt.

A pH measurement of the test item at a concentration of 10% W/w in sterile water.
pH measurement:
Initial reading = 3.02
After 10 minutes = 3.04

For the purpose of this study the test item was used as supplied.
The negative control item, 0.9% w/v sodium chloride solution, was used as supplied.
The positive control item, Ethanol, was used as supplied.

Vehicle:

unchanged (no vehicle)

Controls:

other: Negative and positive controls

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 ml/eye

Duration of treatment / exposure:

10 minutes

Observation period (in vivo):

The holders were incubated, anterior chamber facing forward, at 32 +/- 1 °C for 120 minutes.

Number of animals or in vitro replicates:

3 tested eyes, 3 negative controls and 3 positive controls

Details on study design:

All eyes were macroscopically examined before and after dissection. Only corneas free of damage were used.
The cornea from each selected eye was removed leaving a 2 to 3 mm rim of sclera to facilitate handling. The iris and lens were peeled away from the cornea. The isolated corneas were immersed (epithelial side uppermost) in a dish containing HBSS until they were mounted in Bovine Corneal Opacity and Permeability (BCOP) holders.
The anterior and posterior chambers of each BCOP holder were filled with complete Eagle’s minimum essential medium(MEM) and plugged. The holders were incubated at 32+/- 1 °C for 60 minutes. At the end of the incubation period each comea was examined for defects. Only corneas free of damage were used.
The medium from both chambers of each holder was replaced with fresh complete MEM.
A pre-treatment opacity reading was taken for each cornea using a calibrated opacitometer. The average opacity for all corneas was calculated.
Three corneas with opacity values close to the median value of all corneas were allocated to the negative control. Three corneas were also allocated to the test item and three corneas to the positive control item.
The MEM was removed from the anterior chamber of the BCOP holder and 0.75 mL of the test item or control items were applied to the appropriate corneas. The holders were gently tilted back and forth to ensure a uniform application of the item over the entire cornea. Each holder was incubated, anterior chamber uppermost, at 32 +/- 1 °C for 10 minutes.
At the end of the exposure period the test item and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete MEM. The anterior chamber was refilled with fresh complete MEM. A post-treatment opacity reading was taken and each comea was visually observed. The holders were incubated, anterior chamber facing forward, at 32 +/- 1 °C for 120 minutes.
After incubation the holders were removed from the incubator and a final opacity reading was taken. Each cornea was visually observed.
Following the opacity measurement the permeability of the comeas to sodium fluorescein was evaluated. The medium from the anterior chamber was removed and replaced with 1 mL of sodium fluorescein solution (4 mg/mL). The dosing holes were plugged and the holders incubated, anterior chamber uppermost, at 32 +/- 1 °C for 90 minutes.
After incubation the medium in the posterior chamber of each holder was decanted and retained. 360 µL of medium representing each comea was applied to a designated well on a 96-well plate and the optical density at 492 mn (OD492) was measured using the Anthos 2001 microplate reader.
The corneas were retained after testing for possible conduct of histopathology. Each cornea was placed into a pre-labeled tissue cassette fitted with a histology sponge to protect the endothelial surface. The cassette was immersed in 10% neutral buffered formalin.

Irritation parameter:

in vitro irritation score

Run / experiment:

3 duplicates

Value:

ca. 7

Vehicle controls validity:

not valid

Negative controls validity:

valid

Positive controls validity:

valid

Irritant / corrosive response data:

Individual and mean comeal opacity measurements and individual and mean corneal permeability measurements are given in Table 1 of the attached document.
The condition of each comea post treatment and at the final opacity measurement is given in Table 2 of the attached document.
The in vitro irritancy scores are summarized in the table hereunder.
The comeas treated with the test item were clear post treatment and slightly cloudy post incubation. The corneas treated with the negative control item were clear post treatment and post incubation. The comeas treated with the positive control item were cloudy post treatment and post incubation.

Results from the two test method endpoints, opacity and permeability, were combined in an empirically derived fonnula to generate an In Vitro Irritancy Score.

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 the average change in opacity observed for the negative control comeas. The mean opacity value of each treatment group was then calculated by averaging the corrected opacity values of each cornea for that treatment group.

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 Irritancy Score:

In Vitro Irritancy Score = mean opacity value + (15 x mean OD492 value)

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

The condition of the cornea was visually assessed immediately after rinsing and at the final opacity measurement.

In Vitro Irritancy Scores  Treatment  In vitro Irritancy Score  Test Item 7.0   Negative Control  0.9 Positive Control   32.3

Interpretation of results:

GHS criteria not met

Conclusions:

The eye irritancy potential of the test material was assessed in accordance with OECD Guideline 437. The test item was considered not to be an ocular corrosive or severe irritant. The result of this study has identified the test item as not causing serious eye damage. As such, the test material does not meet the GHS criteria for classification.

Executive summary:

Introduction

A study was performed to assess the ocular irritancy potential of the test item to the isolated bovine cornea.

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 fonnula to generate an In Vitro Irritancy Score (IVIS).

Method

The test item is classified according to the prediction model below:

IVIS CLASSIFICATION

<3 No category. Not requiring classification to UN GHS or EU CLP

> 3 and < 55 No prediction of eye irritation can be made

> 55 Category 1. UN GHS or EU CLP Causes serious eye damage

The In Vitro irritancy scores are summarized as follows:

Test Item: 7.0

Negative Control: 0.9

Positive Control: 32.3

Conclusions

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

The result of this study has identified the test item as not causing serious eye damage.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 July 2014 to 04 August 2014

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)

Specific details on test material used for the study:

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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories U.K. Ltd. Leicestershire UK
- At the start ofthe study the animals weighed 2.29 or 2.35 kg and were twelve to twenty weeks old.
- The animals were individually housed in suspended cages. Free access to mains drinking water and food. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity ofthe study.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):20 ± 3 °C
- Humidity (%):30 - 70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light):12 hours light and 12 hours darkness.
- The animals were provided with environmental enrichment items which were considered not to contain any contaminant ofa level that might have affected the purpose or integrity of the study.

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml/animal

Duration of treatment / exposure:

No wash done

Observation period (in vivo):

1, 24, 48 and 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 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 anesthesia of ocular anesthetic
(two drops of 0.5% tetracaine hydrochloride) was applied to each eye.
A volume of 0.1 mL ofthe 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 six point scale.
Eight hours after test item application, a subcutaneous injection of post-dose analgesia, buprenorphine 0.01 mg/kg and nieloxicam 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.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Individual and group mean scores for ocular irritation are given in Table 1 and Table 2 of the attached document.
No corneal effects were noted during the study.
Iridial inflammation was noted in both treated eyes 1 and 24 hours after treatment and persisted in one treated eye at the 48-Hour observation.
Moderate conjunctival irritation was noted in both treated eyes 1 hour after treatment and at the 24 and 48-Hour observations. Moderate conjunctival irritation persisted in one treated eye with minimal conjunctival irritation noted in the other treated eye at the 72-Hour observation.
Both treated eyes appeared normal at the 7-Day observation.

Other effects:

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. No clinical signs noted.
Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period. Both animals showed expected gain in body weight during the study.

Interpretation of results:

Category 2B (mildly irritating to eyes) based on GHS criteria

Conclusions:

The test item did not cause any damage to the rabbit eye. The individual mean scores for corneal opacity and iris light reflex were respectively 0.00 and 0.5 for all two animals. The individual mean scores for the conjunctivae were 2 for reddening and 1.33 for chemosis for all animals. Therefore, the test item will be classified the following way; GHS classification category 2B, EU CLP classification category 2, and a Canadian WHMIS classification D2B. The classification is due to the fact that during the 24 to 72 hour time period both animals had conjunctival redness equal to 2 and was fully reversible within the 7 days of observation.

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.

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 7-Day observation.

Conclusions

The test item did not cause any damage to the rabbit eye. The individual mean scores for corneal opacity and iris light reflex were respectively 0.00 and 0.5 for all two animals. The individual mean scores for the conjunctivae were 2 for reddening and 1.33 for chemosis for all animals. Therefore, the test item will be classified the following way; GHS classification category 2B, EU CLP classification category 2, and a Canadian WHMIS classification D2B.  The classification is due to the fact that during the 24 to 72 hour time period both animals had conjunctival redness equal to 2 and was fully reversible within the 7 days of observation.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation

The key study for skin irritation was performed on the read-across substance Ethylene glycol, reaction products with polyisobutenyl succinic anhydride and hexadecenyl succinic anhydride, salts with dimethylethanolamine. The justification for read-across is attached in Section 13 of the IUCLID.

A study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method followed the recommendations of the US Environmental Protection Agency (EPA) Guidelines, OECD Guidelines for Testing of Chemicals No. 404 "Acute Dermal Irritation/Corrosion" (adopted 17 July 1992) and Method 84 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 6 7/548/E EC) and Austrialian Toxicology Guidelines.

A single 4-hour, semi-occluded application of the test material to the intact skin of six rabbits produced very slight to well-defined erythema and very slight oedema. Desquamation was noted on day 7 but was considered to be a reversible effect which did not warrant continuation of the observation period.

The test material, produced a primary irritation index of 0.72 in relation to erythema only, and which was fully reversible by the 7-day observation period.

The substance is not classified for skin irritation in accordance with the CLP Regulation 1272/2008.

Eye Irritation

In vitro

A study was performed to assess the ocular irritancy potential of the test item to the isolated bovine cornea.

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 fonnula to generate an In Vitro Irritancy Score (IVIS).

The test item is classified according to the prediction model below:

IVIS CLASSIFICATION

<3 No category. Not requiring classification to UN GHS or EU CLP

> 3 and < 55 No prediction of eye irritation can be made

> 55 Category 1. UN GHS or EU CLP Causes serious eye damage

The In Vitro irritancy scores are summarized as follows:

Test Item: 7.0

Negative Control: 0.9

Positive Control: 32.3

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

The result of this study has identified the test item as not causing serious eye damage.

In vivo

The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit.

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 7-Day observation.

The test item did not cause any damage to the rabbit eye. The individual mean scores for corneal opacity and iris light reflex were respectively 0.00 and 0.5 for all two animals. The individual mean scores for the conjunctivae were 2 for reddening and 1.33 for chemosis for all animals. Based on these results, the test item is considered to be “irritating” to rabbit eye. Ocular reactions were fully reversed by Day 7.

The substance is classified for eye irritation (Category 2) in accordance with the CLP Regulation 1272/2008.

Justification for classification or non-classification

The test material is not irritating to the skin of rabbits based on read-across to data on a surrogate substance. Based on in vivo data, the test material was determined to be irritating to the eye of rabbits in accordance with Regulation EC 1272/2008.