1,12-dodecanediyl bismethacrylate

Endpoint summary

Description of key information

Two reliable studies are available to evaluate the skin and eye irritation potential of an analogue of 1,12 -dodecaldiol bismethacrylate : 1,10 -decanediol diacrylate. 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 vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 March 2013 - 29 April 2013

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:

yes

Remarks:

see below

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 439 (In vitro dermal irritation)

Deviations:

no

Principles of method if other than guideline:

§ Test for direct MTT reduction with the test item: due to a typing error in the study plan, the volume of MTT solution used for the negative control in the preliminary test for direct MTT reduction which is specified in the study plan is incorrect. Indeed, the volume of MTT solution used in this study was 2 mL (as described in CiToxLAB’s SOP) instead of 2.2 mL. This deviation was considered not to have compromised the validity or integrity of the study.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

no

Test system:

human skin model

Source species:

other: reconstructed human epidermis

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. Excess D-PBS was removed by blotting the bottom of the tissue culture insert with absorbent paper.
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 (± 1) hours.

POSITIVE CONTROL
Name: Sodium Dodecyl Sulphate (SDS) at a 5% (w/v) aqueous solution.
The supplier and batch number of the positive control used are documented in the study files.

NEGATIVE CONTROL
Name: Dulbecco’s Phosphate-Buffered Saline (D-PBS).
The supplier and batch number of the negative control used are documented in the study files.

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

Interpretation: see in "any other information on materials and methods"

Control samples:

yes, concurrent vehicle

yes, concurrent positive control

Amount/concentration applied:

Amount(s) applied per tissue: 10 µL ± 2 µL

Duration of treatment / exposure:

Exposure period of 15 minutes, followed by a rinsing.

Duration of post-treatment incubation (if applicable):

MTT-loading after a 42h-incubation period following rinsing. Observation of MTT-> formazan transformation by viable cells, after 3-hour incubation.

Number of replicates:

Triplicate tissues for each tested substances (test item, negative control, positive control).

Irritation / corrosion parameter:

% tissue viability

Value:

112

Vehicle controls validity:

valid

Negative controls validity:

not applicable

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 -minute exposure and a 42-hour recovery period, the relative mean viability of the tissues treated with the test item was 112% with a standard deviation of 6%. As the mean viability was > 50% after the MTT reduction,the results met the criteria for an in vitro classification as non-irritant to skin.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item is considered to be non-irritant to skin.

Executive summary:

The objective of this study was to evaluate the skin irritation potential of the test item using the Episkin^TMreconstructed 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’s 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 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% CO₂in 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).

 

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 -minute exposure and a 42-hour recovery period, the relative mean viability of the tissues treated with the test item was 112% with a standard deviation of 6%. As the mean viability was > 50% after the MTT reduction,the results met the criteria for an in vitro classification as non-irritant to skin.

 

Conclusion

 

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

 

 

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:

12 April 2013 - 27 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:

yes

Remarks:

the animals were acclimated to the study conditions for a period of 4 days instead of at least 5 days.

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: 2590 g to 3130 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: for a period of 4 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: 14 May 2013 to 24 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 performed before 24-hour reading.

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.

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 (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:

other: 3 rabbits

Time point:

other: 24, 48 and 72 h (mean)

Score:

0

Max. score:

4

Reversibility:

other: not applicabke

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

other: 3 rabbits

Time point:

other: 24, 48 and 72 h (mean)

Score:

0

Max. score:

3

Reversibility:

other: not applicabke

Irritation parameter:

iris score

Basis:

other: 3 rabbits

Time point:

other: 24, 48 and 72 h (mean)

Score:

0

Max. score:

2

Reversibility:

other: not applicabke

Irritation parameter:

cornea opacity score

Remarks:

(intensity)

Basis:

other: 3 rabbits

Time point:

other: 24, 48 and 72 h (mean)

Score:

0

Max. score:

4

Reversibility:

other: not applicabke

Irritant / corrosive response data:

In the left treated eye, slight chemosis and slight or moderate 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.

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

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. 405 and the principles of Good Laboratory Practices.

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 and was < 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 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 (i.e. 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 or moderate 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.

 

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 significant eye irritation,

. redness of the conjunctiva: 0.0, 0.0 and 0.0; showing no significant 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)/ read-across:

The objective of this study was to evaluate the skin irritation potential of the test item using the Episkin 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’s standard operating procedures and the principles of Good Laboratory Practice.

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 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.

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

In the main test, all acceptance criteria for the negative and positive controls were fulfilled. The study was therefore considered to be valid.Following a 15 -minute exposure and a 42-hour recovery period, the relative mean viability of the tissues treated with the test item was 112% with a standard deviation of 6%. As the mean viability was > 50% after the MTT reduction,the results met the criteria for an in vitro classification as non-irritant to skin. 

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

In vivo eye irritation test (Papineau 2013)/ read-across:

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. 405 and the principles of Good Laboratory Practices.

The test item was administered to 3 male New Zealand White rabbits.

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 or moderate 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.

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 significant eye irritation,

. redness of the conjunctiva: 0.0, 0.0 and 0.0; showing no significant 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.

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

Based on the available on the analogue 1,10-decanediol diacrylate, no classification of 1,12 -dodecanediol bismethacrylate is required for skin or eye irritation endpoints according to the Regulation EC n°1272/2008.