Isodecyl acrylate

Endpoint summary

Description of key information

Isodecyl acrylate is not irritating for skin and eye, according to the in vitro and in vivo tests performed on this test substance.
However, isodecyl acrylate is considered to be irritating for the respiratory tract.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 August 2012 to 09 January 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes

Test system:

human skin model

Source species:

human

Cell type:

other: skin model EpiDerm SIT (EPI-200).

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm SIT (EPI-200): A three-dimensional human skin model, comprising a reconstructed epidermis with a functional stratum corneum was supplied by MatTek corporation, Ashland, Massachusetts, USA.

Amount/concentration applied:

30 µL of either negative/ positive control or test article.

Duration of treatment / exposure:

The tissues were incubated at 37°C, 5% carbon dioxide for a 35 minute period.
The plates were then removed from the incubator and placed into a sterile hood for the remaining 60 minutes.

Duration of post-treatment incubation (if applicable):

Following treatment, the substances were removed by washing the tissues. The tissues were then placed on the appropriate medium and incubated for 42 hours.

Number of replicates:

3

Details on study design:

At the end of the 42-hour incubation period, tissue viability was assessed by MTT assay. Following rinsing, tissues were placed on 0.3 mL of MTT solution (1 mg/mL) and incubated for 3 hours. Once complete, the tissues were removed from the MTT solution and any resultant colour formed in the tissues by the MTT assay was extracted.
Extraction was achieved by flooding the tissue with 2 mL isopropanol, sealing the plate to avoid any evaporation occurring, and then shaking at 150 rpm for 2 hours, protected from light.
Upon completion of the extraction, each tissue was pierced using a hypodermic needle so that the extract could run through the tissue. Once drained, the tissue was discarded and the extract mixed by shaking at 150 rpm for 15 minutes. Three 200 µL aliquots of each resultant extract were placed into a 96-well plate for spectrophotometric determination of optical density at 570 nm using extraction solution as blank.
Tissue viability was calculated for each tissue as a percentage of mean of the negative control tissues.

Irritation / corrosion parameter:

% tissue viability

Value:

114

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The group mean viability for the test article was 114.2%
The group mean viability for the negative control was 100%
The goup mean viability for the positive control was 3.5%

Other effects:

no

Interpretation of results:

GHS criteria not met

Conclusions:

The test article, Isodecyl Acrylate, was considered not to be irritating to the in vitro skin model EpiDerm SIT (EPI-200).

Executive summary:

This study was conducted to determine whether the test article, Isodecyl Acrylate, causes dermal irritation in the in vitro skin model EpiDerm SIT (EPI-200).

EpiDerm SIT (EPI-200) inserts were treated with test article, negative control (phosphate buffered saline (PBS)) and positive control (5% w/v sodium dodecyl sulphate (SDS)) for 60 minutes. At the end of the treatment period, the tissues were washed with PBS and cell viability was assessed using the MTT assay. The skin irritation potential was classified according to the remaining cell viability obtained after test article treatment.

The group mean viability for the test article was 114.2%, for the negative control was 100% and for the positive control was 3.5%.

The test article, Isodecyl Acrylate, was considered not to be irritant to the in vitro skin model EpiDerm SIT (EPI-200).

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 September 2012 to 11 January 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

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan UK Ltd., Bicester
- Age at study initiation: 13 to 14 weeks
- Weight at study initiation: 2.7 to 3.0 kg
- Housing: Individually housed in cages that conformed to the 'Code of Practice for the Housing and Care of Animals Used in Scientific Procedures'
- Diet (e.g. ad libitum): Global Diet 2930C, ad libitum
- Water (e.g. ad libitum): Mains water, ad libitum
- Acclimation period: 7 to 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15 to 21°C
- Humidity (%): 45%
- Air changes (per hr): 15 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light): Rooms illuminated by fluorescent strip-lights for twelve hours daily

IN-LIFE DATES: From: 18 September 2012 to: 05 October 2012

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

0.1 mL of undiluted test article instilled into the left conjunctival sac of each rabbit (one sentinel rabbit, followed by two further rabbits).

Duration of treatment / exposure:

Ocular reactions were assessed for up to 4 days after treatment.

Observation period (in vivo):

The treated animals were examined for ocular changes approximately 30 minutes, one and four hours after treatment and on Days 2, 3 and 4 (at approximately 24, 48 and 72 hours post-treatment.)

Number of animals or in vitro replicates:

3 rabbits

Details on study design:

The test article did not demonstrate corrosive or severe irritant responses in a validated and accepted ex vivo test designed for the specific assessment of eye corrosivity/irritation (Covance study number 8268255), and did not demonstrate irritant responses in a validated and accepted in vitro test designed for the specific assessment of skin irritation (Covance study number 8268258).
Furthermore, before the first animal was dosed, the pH of the test article was checked, and found to be 5 (i.e. within the acceptable range of pH 2 to 11.5).

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

(3 rabbits)

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

(3 rabbits)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

(3 rabbits)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Remarks:

(3 rabbits)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritant / corrosive response data:

Ocular instillation of the test article provoked practically no initial sting reaction.
There were no ocular responses noted other than minimal conjunctival discharge in one animal at 30 minutes and 1 hour after instillation.

Other effects:

No observations indicative of systemic toxicity or ill health were noted for any rabbits during the course of the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The test article produced a maximum group mean score of 0.6 and was considered to be practically non irritant according to the modified version of the system described by Kay and Calandra.

Executive summary:

This study was conducted to determine the irritancy caused to the rabbit eye following a single instillation of the test article, Isodecyl Acrylate, into the conjunctival sac.

The undiluted test article (0.1 mL) was instilled into one conjunctival sac of each of three New Zealand White rabbits on Day 1. Ocular reactions were assessed for up to 4 days after treatment.

Ocular instillation of the test article provoked practically no initial sting reaction.

There were no ocular responses noted other than minimal conjunctival discharge in one animal at 30 minutes and 1 hour after instillation.

The test article produced a maximum group mean score of 0.6 and was considered to be practically non-irritant according to the modified version of the system described by Kay and Calandra.

The test article did not meet the criteria for classification as irritant according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

 

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 August 2012 to 09 January 2013

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

Species:

other: Bovine

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Bovine corneas supplied by the local abattoir. The eyes were removed after slaughter, completely immersed in Hanks' Balanced Salt Solution (containing penicillin at 100 IU/mL and streptomycin at 100 µg/mL) in a suitably sized container and transported on the same day to the testing facility.
On arrival at the test facility the eyes were carefully examined for defects including increased opacity, scratches and neovascularisation. Only corneas free from such defects were used.
All corneas were preserved in 10% Neutral Buffer Formalin.
The corneas were then loaded onto specially designed holders and filled with pre-warmed Eagle's Minimal Essential Medium (EMEM) (posterior chamber first), ensuring no bubbles formed. The holders were maintained at 32 ± 1°C for at least 1 hour.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

A volume of 0.5 mL of the test article was applied to each of the cornea followed by a 10 minute incubation at 32°C.
The assay was repeated due to the positive controls not meeting the requirements. 1 mL of the test article was applied in the repeat assay.

Duration of treatment / exposure:

The corneas were incubated (horizontally) for 2 hours ± 10 minutes after which the opacities were measured and then the anterior chamber emptied. For the permeability endpoint, 1 mL of sodium fluorescein (4 mg/mL solution) was added to the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes.

Observation period (in vivo):

Not applicable.

Number of animals or in vitro replicates:

Not applicable. Three corneas used for each treatment.

Details on study design:

Fresh corneas, mounted onto specifically designed holders, were treated topically with the test material. Eye corrosion / severe irritation potential was based on the combined effect of the test article on the opacity of the cornea following the treatment and the cornea's ability to resist penetration of a fluorescent dye through the tissue.
The negative control substance was 0.9% sodium cholride solution, the positive control substance was dimethyl formamide.

Irritation parameter:

in vitro irritation score

Run / experiment:

1

Value:

0.37

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The mean opacity reading for the test article was 0.3, for the negative control was 0.0 and for the positive control was 22.3.
The mean group corrected optical density for the test article was 0.002, for the negative control was 0.0 and for the positive control was 0.295.
The IVIS for test article was 0.37, for the negative control was 0.0 and for the positive control was 26.76.
The results are showed in the tables.

Calculated IVIS

 

Test chemical	Mean opacity	Mean permeability	IVIS (mean opacity + (15 × mean permeability))
Negative control	0	0	0
Positive control	22.3	0.295	26.76
Test article	0.3	0.002	0.37

 

Interpretation of results:

GHS criteria not met

Conclusions:

The test article, Isodecyl Acrylate, produced an IVIS score of 0.37 and was considered not to be corrosive or irritating to the eye.

Executive summary:

This study was conducted to determine whether the test article, Isodecyl Acrylate, caused corrosion or severe irritation to excised bovine corneas using the bovine corneal opacity and permeability (BCOP) assay.

A volume of 1 mL of the test article formulation was applied to each of three corneas followed by ten minutes incubation at 32°C. A volume of the negative and positive control was similarly applied to further groups of three corneas. After this incubation, each cornea was washed and the anterior chamber filled with pre-warmed EMEM (without phenol red). The corneas were then incubated for an additional 2 hours 32 ± 1°C. At the end of the post-incubation period the corneas were assessed for opacity.

For the permeability endpoint, sodium fluorescein solution was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following this period, the media in the posterior chamber was removed and three 350 µL aliquots of this media (per cornea) were analysed for optical density at 490 nanometers (OD490).

The mean opacity reading for the test article was 0.3, for the negative control was 0.0 and for the positive control was 22.3.

The mean group corrected optical density for the test article was 0.002, for the negative control was 0.0 and for the positive control was 0.295.

The IVIS for test article was 0.37, for the negative control was 0.0 and for the positive control was 26.76.

The test article, Isodecyl Acrylate, produced an IVIS score of 0.37 and was considered not to be corrosive or irritating to the eye.

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

In vitro skin irritation study, EpiDerm test (OECD 439):

This study was conducted to determine whether the test article, Isodecyl Acrylate, causes dermal irritation in the in vitro skin model EpiDerm SIT (EPI-200). At the end of the treatment period (60 min), the tissues were washed with PBS and cell viability was assessed using the MTT assay. The skin irritation potential was classified according to the remaining cell viability obtained after test article treatment.

The group mean viability for the test article was 114.2%, for the negative control was 100% and for the positive control was 3.5%.

The test article, Isodecyl Acrylate, was considered not to be irritant to the in vitro skin model EpiDerm SIT (EPI-200).

In vivo skin irritation studies (Draize tests):

Two old in vivo studies are available to evaluate the skin irritation potential of isodecyl acrylate.

In both studies, rabbits were treated with isodecyl acrylate during 24 hours. The results showed that the test item induces a mild skin irritation.

In vitro eye irritation study, BCOP test (OECD 437):

This study was conducted to determine whether the test article, Isodecyl Acrylate, caused corrosion or severe irritation to excised bovine corneas using the bovine corneal opacity and permeability (BCOP) assay.

A volume of 1 mL of the test article formulation was applied to each of three corneas followed by ten minutes incubation at 32°C. A volume of the negative and positive control was similarly applied to further groups of three corneas. After this incubation, each cornea was washed and the anterior chamber filled with pre-warmed EMEM (without phenol red). The corneas were then incubated for an additional 2 hours 32 ± 1°C. At the end of the post-incubation period the corneas were assessed for opacity.

For the permeability endpoint, sodium fluorescein solution was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following this period, the media in the posterior chamber was removed and three 350 µL aliquots of this media (per cornea) were analysed for optical density at 490 nanometers (OD490).

The mean opacity reading for the test article was 0.3, for the negative control was 0.0 and for the positive control was 22.3.

The mean group corrected optical density for the test article was 0.002, for the negative control was 0.0 and for the positive control was 0.295.

The IVIS for test article was 0.37, for the negative control was 0.0 and for the positive control was 26.76.

The test article, Isodecyl Acrylate, produced an IVIS score of 0.37 and was considered not to be corrosive or severely irritating to the eye.

 

In vivo eye irritation studies on rabbits:

The key study (Dreher 2013) was conducted to determine the irritancy caused to the rabbit eye following a single instillation of the test article, Isodecyl Acrylate, into the conjunctival sac. The undiluted test article (0.1 mL) was instilled into one conjunctival sac of each of three New Zealand White rabbits on Day 1. Ocular reactions were assessed for up to 4 days after treatment. Ocular instillation of the test article provoked practically no initial sting reaction. There were no ocular responses noted other than minimal conjunctival discharge in one animal at 30 minutes and 1 hour after instillation. The test article produced a maximum group mean score of 0.6 and was considered to be practically non-irritant according to the modified version of the system described by Kay and Calandra.

Two old supporting studies showed the same results on eyes.

Justification for classification or non-classification

Skin irritation

A reliable in vitro skin irritation study showed negative results. Two old studies on rabbits are available and showed skin irritation. However, based on the fact that

-the REACh regulation allows to conclude on skin irritation classification based on in vitro studies only,

-the in vitro study available is a recent study (2013), performed with the actual composition of the registered substance,

-both in vivo studies are old, no details on purity are available, and the exposure conditions are stringent (24h of exposure indeed of 4h in the actual test guideline),

Self-classification : No classification for skin irritation is justified for isodecyl acrylate according to the Regulation EC n°1272/2008.

Mandatory classification: Generic classification of monoacrylate should be applied: skin irrit.2 (H315).

Eye irritation

Based on the negative results obtained in the in vitro and in vivo studies performed on isodecyl acrylate, the test susbtance is not eye irritating.

Self-classification : No classification for eye irritation is justified for isodecyl acrylate according to the Regulation EC n°1272/2008.

Mandatory classification: Generic classification of monoacrylate should be applied: Eye Irrit.2 (H319).

Respiratory irritation

The acute inhalation studies available on isodecyl acrylate showed no respiratory irritation. However, based on the read-across with 2 -ethylhexyl acrylate for inhalation repeated toxicity endpoint, isodecyl acrylate should be classified as irritating for respiratory tract:

Self and mandatory classification : STOT SE 3, H335 "May cause respiratory irritation" according to the Regulation EC n°1272/2008.