Poly(oxy-1,2-ethanediyl), .alpha.,.alpha.'-[(1-methylethylidene)di-4,1-phenylene]bis[.omega.-hydroxy-, mixed acrylates and isononanoates

Endpoint summary

Description of key information

Two in vivo reliable studies (Valin 2013 and Papineau 2013) are available to evaluate the skin and eye irritation potential of 4,4'-lsopropylidenediphenol, ethoxylated, esters with acrylic acid and isononanoic acid. The substance is not irritating for skin and eyes.

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:

09 April 2013 - 03 May 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Due to the inconclusive results obtained in the in vitro study, an in vivo study is required to conclude of the skin irritation potential of 4,4'-Isopropylidenediphenol, ethoxylated, esters with acrylic acid and isononanoic acid.

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:

TEST ANIMALS
- Source: breeder: CEGAV, Argenvilliers, France
- Age at study initiation: 2 to 4 months old on the day of treatment
- Mean body weight at study initiation: 2923 g (range: 2875 g to 3015 g)
- Fasting period before study: no
- Housing: the animals were individually housed in noryl cages
- Diet: 110 pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 23 April 2013 to 03 May 2013.

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 mL/flank.

Duration of treatment / exposure:

3 min, 1 h, 4 h.

Observation period:

1, 24, 48 and 72 h after removal of the dressing; if relevant, daily until reversibility of reactions

Number of animals:

Three males

Details on study design:

TEST SITE
- Area of exposure: The test item was placed on a gauze pad, which was then applied to a skin area of approximately 6 cm2.
A dosage-volume of 0.5 mL/flank was used.
During each exposure time, a dry gauze pad was applied to the opposite flank, in order to check that no alteration of the skin is induced by the semi-occlusive dressing and restraining bandage.
The untreated flank acted as control.
The gauze pad was held in place by a non-irritating semi-occlusive dressing and a restraining bandage. After required period of contact with the skin, the dressing was removed.
After removal of the dressing, any residual test item was wiped off by means of a dry cotton pad.

- Coverage: 6 cm2
- Type of wrap if used: gauze pad held in place by a non-irritation semi-occlusive dressing and a restraining bandage.

REMOVAL OF TEST SUBSTANCE
- Washing: using a dry cotton pad
- Time after start of exposure: at removal of each dressing (see Duration of exposure)

SCORING SYSTEM:

- Erythema and eschar formation:
0 no erythema
1 very slight erythema (barely perceptible)
2 well-defined erythema
3 moderate to severe erythema
4 severe erythema (beet redness) to slight eschar formation (injuries in depth)

- Edema formation:
0 no edema
1 very slight edema (barely perceptible)
2 slight edema (edges of area well-defined by definite raising)
3 moderate edema (raised approximately 1 millimeter)
4 severe edema (raised more than 1 millimeter and extending beyond area of exposure)

- Any other lesions were noted

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: Day 4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: Day 4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

After a 4-hour exposure, a very slight erythema was noted in 2/3 animals from day 1 to day 3.
No edema was observed in any animals.
Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:
.            erythema: 0.0, 0.7, 0.7; showing no significant inflammation,
.            edema: 0.0, 0.0; 0.0; showing no inflammation.

Other effects:

No unscheduled deaths occurred during the study and no clinical signs indicative of systemic toxicity were noted in any animals.
The body weight of the animals was unaffected by the test item treatment.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, the test item was slightly irritant when applied topically to rabbits.
However, the test item should not be classified as irritating to skin according to the criteria of CLP Regulation.

Executive summary:

The objective of this study was to evaluate the potential corrosive and irritant properties of the test item following dermal application on rabbits.

This study was conducted in compliance with OECD Guideline No. 404 and the principles of Good Laboratory Practices.

Methods

The test item was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit.

After the 4-hour application, since the mean value from grading at 24, 48 and 72 hours after patch removal was < 2.3 for erythema or for edema, the test item was applied on the skin of two otheranimalsfor 4 hours.

A dosage-volume of 0.5 mL/flank was used.

The test item was placed on a gauze pad, which was then applied to a skin area of approximately 6 cm². The gauze pad was held in place by a non-irritation semi-occlusive dressing and a restraining bandage.After required period of contact with the skin, the dressing was removed.

Each animal was observed at least once a day for mortality and clinical signs. For each exposure period, cutaneous reactions were evaluated approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the reversibility of cutaneous reactions. The mean values of the scores for erythema and edema were calculated for each animal. Body weight was recorded on the day of treatment and at the end of the evaluation of cutaneous reactions.

On completion of the observation period, the animals were used subsequently for the evaluation of the ocular irritation potential of the same test item.

 

Results

No unscheduled deaths occurred during the study and no clinical signs indicative of systemic toxicity were noted in any animals.

The body weight of the animals was unaffected by the test item treatment.

After a 4-hour exposure, a very slight erythema was noted in 2/3 animals from day 1 to day 3.

No edema was observed in any animals.

Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:

.            erythema: 0.0, 0.7, 0.7; showing no significant inflammation,

.            edema: 0.0, 0.0; 0.0; showing no inflammation.

 

Conclusion

Under the experimental conditions of this study, the test item was slightly irritant when applied topically to rabbits.

However, the test item should not be classified as irritating to skin according to the criteria of CLP Regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 April 2013 - 17 May 2013

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

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: breeder: CEGAV, Argenvilliers, France.
- Age at study initiation: 2 to 4 months old on the day of treatment
- Mean body weight at study initiation: the animals had a mean body weight of 3248 g (range: 3055 g to 3360 g).
- Fasting period before study: no
- Housing: individually housed in noryl cages
- Diet: 110 pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: 07 May 2013 to 17 May 2013

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated right eye served as a control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 mL/animal

Duration of treatment / exposure:

Not applicable: single application followed by rinsing before 24-hour reading in 2/3 animals.

Observation period (in vivo):

1, 24, 48 and 72 h

Number of animals or in vitro replicates:

three males

Details on study design:

REMOVAL OF TEST SUBSTANCE: yes in 2/3 animals.

SCORING SYSTEM: Draize scale.

- Conjunctival chemosis (lids and/or nictitating membranes):
0 no swelling
1 any swelling above normal
2 obvious swelling with partial eversion of lids
3 swelling with lids about half-closed
4 swelling with lids more than half-closed

- Conjunctival redness (refers to the most severe reading for the palpebral and bulbar conjunctivae, excluding the cornea and iris):
0 blood vessels normal
1 a number of blood vessels definitely hyperemic (injected)
2 diffuse, crimson colour, individual vessels not easily discernible
3 diffuse, beefy red

- Iris lesions
0 normal
1 markedly deepened rugae, congestion, swelling, moderate circum-corneal hyperemia,or injection, any or a combination of any there findings, but iris still reacting to light (sluggish reaction is positive)
2 no reaction to light, haemorrhage, gross destruction (any or all of these)

- Corneal intensity of opacity (direct examination and, if necessary, with an UV lamp)
0 no ulceration or opacity
1 scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible
2 easily discernible translucent area, details of iris slightly obscured
3 nacreous areas, no details of iris visible, size of pupil barely discernible
4 opaque cornea, iris not discernible through the opacity

- Corneal area of opacity (direct examination and, if necessary, with an UV lamp)
1 one quarter (or less) but not zero
2 greater than one quarter but less than a half
3 greater than one half but less than three quarters
4 greater than three quarters up to whole area

- Any other lesions observed were noted

TOOL USED TO ASSESS SCORE: UV lamp after instillation of 0.5% sodium fluorescein solution

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

cornea opacity score

Remarks:

(intensity)

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

In the left treated eye, slight chemosis and slight redness of the conjunctiva were observed in all animals on day 1. Then, no ocular reactions were observed in any animals until the end of the observation period (day 4).
Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:
- chemosis: 0.0, 0.0 and 0.0; showing no eye irritation,
- redness of the conjunctiva: 0.0, 0.0 and 0.0; showing no eye irritation,
- iris lesions: 0.0, 0.0 and 0.0; showing no eye irritation,
- corneal opacity: 0.0, 0.0 and 0.0; showing no eye irritation.

Other effects:

No unscheduled deaths occurred during the study and no clinical signs were noted in any animals.
The body weight of the animals was unaffected by the test item treatment.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, the test item was very slightly irritant when administered by ocular route to rabbits.

Executive summary:

The objective of this study was to evaluate the potential irritant properties of the test item for the eye following a single administration to rabbits.

This study was conducted in compliance with OECD Guideline No. 405and the principles of Good Laboratory Practices.

Methods

 

The test item was first administered to a single male New Zealand White rabbit.

 

As mean value from grading at 24, 48 and 72 hours after instillation was < 2 for conjunctival edema (chemosis) and for conjunctival redness; < 1 for iris lesion and for corneal opacity, the test item was administered in the left eye of two other animals.

 

The test item was administered inthe conjunctival sac of the left eye. The right eye remained untreated and served as control.

A dosage-volume of 0.1 mL/animal was used.

For all animals,a local anesthetic was used prior to treatment.

Before 24-hour reading, both eyes of two males were rinsed with a sterile isotonic saline solution (0.9% NaCl).

Each animal was observed at least once a day for mortality and clinical signs. Ocular reactions were observed approximately 1, 24, 48 and 72 hours after the administration and then daily until the end of the observation period. The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal. Body weight was recorded on the day of treatment and at the end of the evaluation of ocular reactions.

On completion of the observation period, the animals were sacrificed then discarded without macroscopic post-mortem examination.

 

Results

 

No unscheduled deaths occurred during the study and no clinical signs were noted in any animals.

The body weight of the animals was unaffected by the test item treatment.

In the left treated eye, slight chemosis and slight redness of the conjunctiva were observed in all animals on day 1. Then, no ocular reactions were observed in any animals until the end of the observation period (day 4).

 

Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:

- chemosis: 0.0, 0.0 and 0.0; showing no eye irritation,

- redness of the conjunctiva: 0.0, 0.0 and 0.0; showing no eye irritation,

- iris lesions: 0.0, 0.0 and 0.0; showing no eye irritation,

- corneal opacity: 0.0, 0.0 and 0.0; showing no eye irritation.

 

Conclusion

 

Under the experimental conditions of this study, the test item was very slightly irritant when administered by ocular route to rabbits.

 

Therefore, the test item should not be classified as irritating to the eyes according to the criteria of CLP Regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vitro skin irritation test (Valin 2013):

The objective of this study was to evaluate the skin irritation potential of the test item using the EpiskinTM reconstituted human epidermis model.

The study design was based upon international guidelines (OECD Guideline No. 439 and Commission Regulation (EC) No. 761/2009, B.46).

In the main test, the test item and both the negative and positive controls were topically applied on triplicate tissues and incubated at room temperature for 15 (± 1) minutes. At the end of the treatment period, each tissue was rinsed with D-PBS and incubated for 42 (± 1) hours at, 5% CO2 in a humidified incubator. The cell viability was then assessed by means of the colourimetric MTT reduction assay. Relative viability values were calculated for each tissue and expressed as a percentage of the mean viability of the negative control tissues which was set at 100% (reference viability).

In the preliminary tests, the test item was found not to have direct MTT reducing properties or a colouring potential.

All acceptance criteria for the negative and positive controls were fulfilled. The study was therefore considered to be valid.Following a 15 minutes exposure and a 42-hour recovery period, the relative mean viability of the tissues treated with the test item was 52% with a standard deviation of 5%. According to the OECD Guideline No. 439, these results are considered as equivocal since the relative mean viability was found equal to 50 ± 5% and since the relative viability data between replicates were non-concordant. Indeed two out of the three replicates had viability values upper the positivity threshold of 50% and the third one had a viability value below. No second test was conducted in this study and therefore these results remained equivocal.

The irritancy potential of the test item is considered to be equivocal to skin. Due to these equivocal results, no classification could be issued.

In vivo skin irritation test (Papineau 2013):

The objective of this study was to evaluate the potential corrosive and irritant properties of the test item following dermal application on rabbits (OECD 404). The test item was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit. After the 4-hour application, since the mean value from grading at 24, 48 and 72 hours after patch removal was < 2.3 for erythema or for edema, the test item was applied on the skin of two other animals for 4 hours. A dosage-volume of 0.5 mL/flank was used. The test item was placed on a gauze pad, which was then applied to a skin area of approximately 6 cm2. The gauze pad was held in place by a non-irritation semi-occlusive dressing and a restraining bandage. After required period of contact with the skin, the dressing was removed. Each animal was observed at least once a day for mortality and clinical signs. For each exposure period, cutaneous reactions were evaluated approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the reversibility of cutaneous reactions. The mean values of the scores for erythema and edema were calculated for each animal. Body weight was recorded on the day of treatment and at the end of the evaluation of cutaneous reactions. No unscheduled deaths occurred during the study and no clinical signs indicative of systemic toxicity were noted in any animals. The body weight of the animals was unaffected by the test item treatment. After a 4-hour exposure, a very slight erythema was noted in 2/3 animals from day 1 to day 3. No edema was observed in any animals. Under the experimental conditions of this study, the test item was slightly irritant when applied topically to rabbits.

In vivo eye irritation test (Papineau 2013):

The objective of this study was to evaluate the potential irritant properties of the test item for the eye following a single administration to rabbits (OECD 405). The test item was first administered to a single male New Zealand White rabbit. As mean value from grading at 24, 48 and 72 hours after instillation was < 2 for conjunctival edema (chemosis) and for conjunctival redness; < 1 for iris lesion and for corneal opacity, the test item was administered in the left eye of two other animals. The test item was administered inthe conjunctival sac of the left eye. The right eye remained untreated and served as control. A dosage-volume of 0.1 mL/animal was used.

Each animal was observed at least once a day for mortality and clinical signs. Ocular reactions were observed approximately 1, 24, 48 and 72 hours after the administration and then daily until the end of the observation period. The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal. Body weight was recorded on the day of treatment and at the end of the evaluation of ocular reactions. No unscheduled deaths occurred during the study and no clinical signs were noted in any animals. The body weight of the animals was unaffected by the test item treatment. In the left treated eye, slight chemosis and slight redness of the conjunctiva were observed in all animals on day 1. Then, no ocular reactions were observed in any animals until the end of the observation period (day 4).Under the experimental conditions of this study, the test item was very slightly irritant when administered by ocular route to rabbits.

Justification for classification or non-classification

No damage or irritation on the skin and eye were observed in the rabbit studies, 4,4'-lsopropylidenediphenol, ethoxylated, esters with acrylic acid and isononanoic acid is not irritating for skin and eyes. No classification is expected for skin and eye irritation endpoints, according to the Regulation EC n°1272/2008.