2-hydroxy-3-phenoxypropyl acrylate

Endpoint summary

Description of key information

Irritating potential of 2 -Hydroxy-3 -phenoxy acrylate was evaluated by in vitro tests.

Negative results were obtained for skin irritation in the vitro study (OECD 439, with a mean cell viability of 79%) and in the in vivo study (OECD 404).

However, the substance is severely irritating when administered into the eyes of rabbits.

Based on these results, 2 -Hydroxy-3 -phenoxy acrylate is considered to be not irritating for skin, but severely irritating to the eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

June 1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

not GLP

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Gaukler, Offenbach/Main, FRG
- Age at study initiation: not specified
- Weight at study initiation: 2.89 - 3.03 kg
- Housing: individually
- Diet (e.g. ad libitum): about 130 g per day per animal
- Water (e.g. ad libitum): about 250 ml per day per animal
- Acclimation period: at least 8 days bafore the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): air-conditionned without details
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

at least 15 hours before the beginning of the study

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated skin sites of the same animal

Amount / concentration applied:

Application of 0.5 ml of the undiluted test substance to the patch test (2.5 x 2.5)

Duration of treatment / exposure:

4h

Observation period:

72h

Number of animals:

1 male and 2 females

Details on study design:

Application site : upper third of the back or flanks
Removal of the test substance at the end of exposure period with lutrol and lutrom/water (1:1)
Readings 30-60 minutes after removal of the test patches and 24, 48, 72 hours after the beginning of application.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 intact animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 intact animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

There is no indication of skin irritation (no erythema and no edema) at 24 hr after exposure.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on this study on rabbits, the test item is considered to be not irritating to the skin.

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.

The test item was applied for 4 hours to a three rabbits with intact skin. 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. After a 4-hour exposure (three animals), no erythema or edema were observed at 24 hours after exposure.

Based on this study on rabbits, the test item is considered to be not irritating to the skin.

 

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 April 2015 - 05 June 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: EU Method B.46 (In vitro dermal irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 439 (In vitro dermal irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

other: reconstructed human epidermis

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiskinTM Model Kit (0.38 cm2 tissues) supplied by SkinEthic Laboratories, Lyon, France.
Medium and Incubation T°C: 37°C

REMOVAL OF TEST SUBSTANCE
- Rinsing: At the end of the treatment period, each tissue was removed from the well of the treatment plate, and rinsed with D-PBS. Rinsing was achieved by gently filling and emptying several times each tissue with D PBS to gently remove any residual test or control items. The rinsed tissues were transferred to the second column of 3 wells containing 2 mL of maintenance medium in each well and the plates were incubated at +37°C, 5% CO2 in a humidified incubator for 42 hours.

POSITIVE CONTROL
Name: Sodium Dodecyl Sulphate (SDS) at a 5% (w/v) aqueous solution.

NEGATIVE CONTROL
Name: Dulbecco’s Phosphate-Buffered Saline (D-PBS).

SCORING SYSTEM:
- Optical density (OD) was measured at 570 nm:
Relative mean viability (%) = 100 x mean cOD(test item) / mean cOD(negative control)
where:
- mean cOD Negative Control = mean ODNC – mean ODblank
- mean cOD Test Item = mean ODTI – mean ODblank

Control samples:

yes, concurrent no treatment

yes, concurrent positive control

Amount/concentration applied:

Amount applied per tissue: 10 +/- 2 µL.

Duration of treatment / exposure:

Exposure period of 15 minutes, followed by rinsing
MTT-loading after a 42h-incubation period following rinsing. Observation of MTT-> formazan transformation by viable cells.

Number of replicates:

Triplicate

Irritation / corrosion parameter:

% tissue viability

Value:

79

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

In the preliminary tests, the test item was found not to have direct MTT reducing properties or colouring potential.
 
Main test: 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 42 hours of recovery period, the relative mean viability of the tissues treated with the test item was 79% with a standard deviation of 15%.
As the mean viability was > 50% after the MTT reduction, the IL-1a concentrations inculture media samples retained from the three negative controls and the three test item-treated tissues were analyzed by ELISA.
The mean IL-1a concentration for treated tissues was 23.8 pg/mL. Due to this value being below 60 pg/mL, the results met the criteria for an in vitro classification as non-irritant to skin.

Other effects:

no

Interpretation of results:

GHS criteria not met

Conclusions:

In vitro, the test item is considered to be non-irritant to the skin.

Executive summary:

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

The study design was based upon international guidelines (OECD Guideline No. 439 and Commission Regulation (EC) No. 761/2009, B.46). The study was conducted in compliance with CiToxLAB France standard operating procedures and the principles of Good Laboratory Practice.

 

Methods

Preliminary tests were performed to detect the ability of the test item to directly reduce MTT as well as its colouring potential.

Following the preliminary tests, the skin irritation potential of the test item was tested in the main test. The test item and both the negative and positive controls were applied topically on triplicate tissues and incubated at room temperature for 15 minutes. At the end of the treatment period, each tissue was rinsed with D-PBS and incubated for 42 hours at 37°C, 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 addition, the concentration of the inflammatory mediator IL-1awas evaluated in the culture medium retained following the 42-hour recovery period. This quantification, based on an ELISA assay, was performed since the mean relative viability of the test item-treated tissues was > 50% following the MTT reduction assay.

 

Results

Preliminary tests

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

 

Main test

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 42 hours of recovery period, the relative mean viability of the tissues treated with the test item was 79% with a standard deviation of 15%.

As the mean viability was > 50% after the MTT reduction, the IL-1a concentrations inculture media samples retained from the three negative controls and the three test item-treated tissues were analyzed by ELISA.

The mean IL-1a concentration for treated tissues was 23.8 pg/mL. Due to this value being below 60 pg/mL, the results met the criteria for an in vitro classification as non-irritant to skin.

 

Conclusion

Under the experimental conditions of this study, the test item is considered to be non-irritant to skin.

According to the results of this study, the classification of the test item should be:

.            not classified (Directive 67/548/EEC) and no category (Regulation (EC) No. 1272/2008).

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:

June / July 1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

not GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1981

Deviations:

no

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Gaukler, Offenbach/Main, FRG
- Age at study initiation: not specified
- Weight at study initiation: 2.86 - 3.14 kg
- Housing: individually
- Diet (e.g. ad libitum): about 130 g per day per animal
- Water (e.g. ad libitum): about 250 ml per day per animal
- Acclimation period: at least 8 days bafore the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): air-conditionned without details
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.1 ml

Observation period (in vivo):

1h, 24h, 48h, 72h, 8d, 15d, 21d after application

Number of animals or in vitro replicates:

3 rabbits

Details on study design:

Single application to the conjunctival sac of the right eyelid ; the substance was not washed out.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 21d

Remarks on result:

other: individual mean results: 1-1-1

Irritation parameter:

iris score

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 21d

Remarks on result:

other: individual mean results: 0-0.67-0.33

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: 21d

Remarks on result:

other: individual mean results: 2-2-2

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

2.3

Max. score:

4

Reversibility:

fully reversible within: 21d

Remarks on result:

other: individual mean results: 2.3-2.3-2.3

Irritant / corrosive response data:

The test item showed signs of eye irritation, not fully reversible in 21 days.

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Under the experimental conditions of this study, the test item was severely irritant with irreversible effects 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.

The test item was administered to 3 rabbits. Ocular reactions were observed approximately 1, 24, 48 and 72 hours, 7, 14 and 21 days after the administration.

Local clinical signs were observed in rabbits: loss of cornea tissue (2 animals), pupil contracted (all), suppuration (2 animals), discharge of blood (2 animals), marginal vascularization of the cornea (2 animals), small retractions in the eyelids (all), bloody incrustation (1 animal).

Mean scores calculated for the 3 rabbits over 24, 48 and 72 hours were: 1 for corneal opacity (individual mean score: 1-1-1), 2.3 for chemosis (2.3-2.3-2.3), 0.33 for iritis (0-0.67-0.33) and 2 for redness of the conjunctiva (2-2-2).

The effects on cornea and conjunctiva are not fully reversible into 21 days.

 Under the experimental conditions of this study, the test item was severely irritant with irreversible effects when administered by ocular route to rabbits.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vitro skin irritation study (Gerbeix 2015a)

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

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

The test item and both the negative and positive controls were applied topically on triplicate tissues and incubated at room temperature for 15 minutes. At the end of the treatment period, each tissue was rinsed with D-PBS and incubated for 42 hours at 37°C, 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 addition, the concentration of the inflammatory mediator IL-1awas evaluated in the culture medium retained following the 42-hour recovery period. This quantification, based on an ELISA assay, was performed since the mean relative viability of the test item-treated tissues was > 50% following the MTT reduction assay.

In the preliminary tests, the test item was found not to have direct MTT reducing properties or 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 42 hours of recovery period, the relative mean viability of the tissues treated with the test item was 79% with a standard deviation of 15%. As the mean viability was > 50% after the MTT reduction, the IL-1a concentrations inculture media samples retained from the three negative controls and the three test item-treated tissues were analyzed by ELISA. The mean IL-1a concentration for treated tissues was 23.8 pg/mL. Due to this value being below 60 pg/mL, the results met the criteria for an in vitro classification as non-irritant to skin.

Under the experimental conditions of this study, the test item is considered to be non-irritant to skin.

In vivo skin irritation study (Kirsch 1989) :

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

The test item was applied for 4 hours to a three rabbits with intact skin. 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. After a 4-hour exposure (three animals), no erythema or edema were observed at 24 hours after exposure.

Based on this study on rabbits, the test item is considered to be not irritating to the skin.

In vivo eye irritation study in rabbit (Kirsch 1989):

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.

The test item was administered to 3 rabbits. Ocular reactions were observed approximately 1, 24, 48 and 72 hours, 7, 14 and 21 days after the administration.

Local clinical signs were observed in rabbits: loss of cornea tissue (2 animals), pupil contracted (all), suppuration (2 animals), discharge of blood (2 animals), marginal vascularization of the cornea (2 animals), small retractions in the eyelids (all), bloody incrustation (1 animal).

Mean scores calculated for the 3 rabbits over 24, 48 and 72 hours were: 1 for corneal opacity (individual mean score: 1-1-1), 2.3 for chemosis (2.3-2.3-2.3), 0.33 for iritis (0-0.67-0.33) and 2 for redness of the conjunctiva (2-2-2).

The effects on cornea and conjunctiva are not fully reversible into 21 days.

Under the experimental conditions of this study, the test item was severely irritant with irreversible effects when administered by ocular route to rabbits.

In vitro eye irritation study (Gerbeix 2015b)

The objective of this study was to evaluate the potential irritant and corrosive properties of the test item to the eye. The Bovine Corneal Opacity and Permeability (BCOP) test method (OECD 437) can identify chemicals inducing serious eye damage and chemicals not requiring classification for eye irritation or serious eye damage.

Three corneas were used for each treated series (test item, positive control and negative control). The test item was applied undiluted, in a single experiment using a treatment time of 10 minutes and using the closed-chamber method. At the end of the incubation period, the optical density of the solution from the posterior chamber of each holder was measured in order to determine the permeability of the cornea. Each cornea was then observed for opaque spots and other irregularities.

Fluorescein fixation was observed on the corneas treated with the test item. All acceptance criteria were fulfilled. The study was therefore considered as valid.

The mean In Vitro Irritancy Score (IVIS) of the test item-treated corneas was: 3. 

As the test item induced a mean IVIS = 3, the test item was considered as a test chemical not requiring classification for eye irritation or serious eye damage.

Justification for classification or non-classification

Based on the available data, 2 -Hydroxy-3 -phenoxy acrylate should be classified as eye corrosive (Eye Dam 1) according to the Regulation EC n°1272/2008.