reaction products of 2-Propenoic acid (2 moles) and Neopentylglycol hydroxypivalate (1 mole)

Endpoint summary

Description of key information

The irritating potential of the registered substance has been evaluated. Based on the rabbit study, the registered substance is considered to be slightly skin irritating but not classified according to the CLP and GHS Regulations. Based on the BCOP, the registered substance is considered to be not eye irritating.

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:

November - December 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Highgate farm, market Rasen, Lincolnshire, England.
- Age at study initiation: at least 9 weeks
- Weight at study initiation: 2.5-2.8 kg
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: > 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19+/-2
- Humidity (%): 30-70
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

Approximately 0.5 ml of the test substance was applied under a 2-ply 25 mm x 25 mm porous gauze pad to each intact skin site on each animal.

Duration of treatment / exposure:

32 min, 60 min, 4h

Observation period:

Examination of the treated skin was made on Day 1 (ie immediately after removal of the dressings following the three minute and one hour exposure or approximately 60 minutes after removal of the dressings following the four hour exposure) and on Day 2, 3 and 4 (equivalent to approximately 24, 48 and 72 hours after exposure). Additional observations were made for all animals on Days 5 and 6, for 2 animals on Day 7 and for one animal on Day 8 through 12.

Number of animals:

3

Details on study design:

Approximately 24 hours prior to application of the test substance, hair was removed with electric clippers from the dorso-lumbar region of each rabbit exposing an area of skin approximately 100 mm x 100 mm.
To help to clarify the irritant potential of the test substance, initially only one animal was treated. With this animal, the test substance was applied to 3 treatment sites and the exposure period was varied between treatment sites. On the basis of initial results from this phase of the study, 2 further animals were exposed to the test substance for 4 hours.
Each treatment site was covered with Elastoplast elastic adhesive dressing for up to 4 hours. only during the 3 minute exposure period was the initial animal restrained. For the 60 minute and 4 hour expsure, none of the animals were restrained but were returned to their cages immediately after treatment.
At the end of the exposure periods, the semi-occlusive dressing and gauzer pad were removed and the treatment sites washed with warm water (34-37°C) to remove any residual test substance. The treated area was blotted dry with absorbent paper

All animals were observed daily for signs of ill health or toxicity.

Irritation parameter:

erythema score

Remarks:

(4h exposure)

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

1.43

Max. score:

4

Reversibility:

fully reversible within: 12 days

Remarks on result:

other: Mean individual scores: 1.3-2-1

Irritation parameter:

edema score

Remarks:

(4h exposure)

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0.87

Max. score:

4

Reversibility:

fully reversible within: 6 days

Remarks on result:

other: Mean individual scores: 0.3-1.3-1

Irritant / corrosive response data:

Very slight erythema was elicited from Day 2 following a single semi-occlusive application of test material to intact skin for 3 and 60 minutes. reactions had resolved completely by Day 4 and 3, respectively.
Very slight to well-defined erythema with slight oedema was observed in all 3 animals following the 4 hour exposure. reactions were first noted approximately an hour after bandage removal and had resolved completely by either Day 6, 7 or 12.
The mean scores for these reactions at approximately 24, 48 and 72 hours after administration, calculated separately for each animal are : 1.3, 2 and 1 for erythema, and 0.3,1.3 and 1 for oedema.

Other effects:

There were no signs of toxicity or ill health in any rabbit during the observation period.

Interpretation of results:

GHS criteria not met

Remarks:

Not skin irritating.

Conclusions:

Based on these results, the test material is considered to be slightly skin irritating.

Executive summary:

A study was performed to assess the skin irritation potential of test material to the rabbit. The method foolowed was that described in OECD test guideline n°404.

Three rabbits were each administered a single dermal dose of 0.5 ml of the test substance an observed for up to 12 days. The effects of a 3 minutes, 60 minutes and 4 hours exposure were initially investigated using a screen animal.

Very slight dermal irritation was elicited following a single semi-occlusive application of test material to intact rabbit skin for 3 and 60 minutes.

A single semi-occlusive application of test material to intact rabbit skin for 4 hours alicited slight to well-defined dermal irritation in all animals.

The mean scores for these reactions at approximately 24, 48 and 72 hours after administration, calculated separately for each animal are : 1.3, 2 and 1 for erythema, and 0.3,1.3 and 1 for oedema.

Based on these results, the test material is considered to be slightly skin irritating.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 December 2016 - 02 February 2017

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 i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: bovine cornea

Strain:

other: not applicable

Details on test animals or tissues and environmental conditions:

Species: bovine cattle.
Origin: bovine eyes were obtained from freshly slaughtered cattle at the abattoir EVA, Saint Pierre sur Dives, France.
Age: as French Authorities avoid the use of any organs from the head of bovines aged more than 12 months, bovine cattle were up to 12 months old (typically, 5 to 8 months old).
Reason for choice: bovine corneas are recommended by Regulatory Authorities for this type of study. They are adapted for the evaluation of potential ocular irritants since they are part of the target organ.
Transport from Supplier to CiToxLAB France: the eyes were transported to CiToxLAB France at ambient temperature, immerged in buffered Hanks medium containing an antibiotic [Hank’s Balanced Salts Solution (HBSS) plus penicillin/streptomycin (100 units/100 µg/mL final)]. A container with smooth internal surfaces was used for the transport to avoid damage to the corneas.
Upon arrival at CiToxLAB France, the selection and preparation of corneas was performed as soon as possible. At each step of the preparation procedure, care was taken to avoid touching the corneas in order not to damage them.

Selection: a careful macroscopic examination was performed on all eyes to detect the presence of any defects (opacity, scratches, pigmentation, etc). Any eyes with defects were discarded. The examination was performed under a lamp, using HBSS in order to keep the eyes moistened and shiny. Particular attention was paid to the corneas and the eyes were swiveled in order to observe the fringe areas and any scratches directly under the light.
Preparation of the selected corneas: the tissues surrounding the eyeball were carefully pulled away and the cornea, surrounded by approximately 2 to 3 mm of sclera, was dissected out. The isolated corneas were stored in HBSS until all corneas had been prepared.
Washing of the corneas: the corneas were washed for 15 minutes, three times, in HBSS plus penicillin/streptomycin (100 units/100 µg/mL final) at room temperature. The corneas were used within a maximum of 24 hours.
Storage of the corneas: as the corneas were not used immediately, they were stored after washing. Each cornea was stored individually in 12 mL of M199 medium containing 5% dextran, plus penicillin/streptomycin, at +4°C, for a maximum of 24 hours before use.

(Pre)Incubation T°C: 32°C
Dates of experimental phase: from 30 January 2017 to 02 February 2017

Vehicle:

unchanged (no vehicle)

Controls:

other: in vitro negative and positive controls

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL

VEHICLE
As the test item is a non-surfactant liquid, it was tested undiluted (i.e. in its original form). Thus, no vehicle was used.

Duration of treatment / exposure:

Exposure period of 10 minutes (± 30 seconds), followed by rinsing

Duration of post- treatment incubation (in vitro):

Opacity measurement:
- before treatment
- after 2-hour incubation in water
Permeability measurement:
- after 90-min incubation in water (and other procedures), following the 2nd opacity measurement

Number of animals or in vitro replicates:

Triplicate corneas for each tested substance (test item, negative control, positive control)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Rinsing: the anterior part of the eye was emptied and then rinsed 5 times with pre-warmed cMEM containing phenol red

NEGATIVE CONTROL:
As the test item was tested undiluted (i.e. in its original form), 0.9% Sodium Chloride (0.9% NaCl) was used as negative control.

Since several test items were assayed concurrently, the negative control was shared.

POSITIVE CONTROL:
As the test item was tested using a 10-minute treatment, the positive control was Absolute Ethanol. It was used neat and sampled on the day of use.

Since several test items were assayed concurrently, the positive control was shared.

SCORING SYSTEM/TOOL
- Opacity:
Using an opacitometer
The average change in opacity during exposure is determined. It is corrected by subtracting the average negative control value from values in positive control and test item.
- Permeability:
Using a spectrophotometer: optical density (OD) at 490 nm wavelength
The optical density is corrected by subtracting the average negative control value from values in positive control and test item.
- Scoring:
In vitro irritancy score (IVIS) = Corrected Opacity + (15 x Corrected OD)

Interpretation: see below

Irritation parameter:

in vitro irritation score

Run / experiment:

treated (10 min)

Value:

1

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

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: 1.
 

Macroscopic examination: No notable opaque spots or irregularities were observed on corneas treated with the test item.

Interpretation of results:

GHS criteria not met

Remarks:

Not eye irritating.

Conclusions:

The test item was identified as not requiring classification for eye irritation or serious eye damage.

Executive summary:

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 can identify chemicals inducing serious eye damage and chemicals not requiring classification for eye irritation or serious eye damage.

The design of this study was based on the guideline OECD Guideline 437 and the study was performed in compliance with CiToxLAB France standard operating procedures and with the OECD Principles of Good Laboratory Practice.

 

Corneas obtained from freshly slaughtered calves were mounted in corneal holders. Both chambers of the corneal holder were filled with complemented MEM culture media (cMEM) and pre-incubated for 1 hour and 5 minutes (± 5 minutes) at +32°C.

A single experiment was performed using three corneas for each treated series (test item, positive control and negative control).

Before the treatment, a first opacity measurement was performed on each cornea using an opacitometer.

The test item was applied undiluted in a single experiment using a treatment time of 10 minutes and using open-chamber method. Negative and positive controls were applied using the same treatment time and using the closed-chamber method. At the completion of the treatment period, all items were removed from the front opening of the anterior chamber and the epithelia were rinsed.

The corneas were then incubated for 2 hours (± 10 minutes) at +32°C before a second opacity measurement was performed.

After the second opacity measurement, the medium of the anterior chamber was removed and filled with a fluorescein solution. The holders were then incubated vertically for 90 minutes (± 5 minutes) at +32°C.

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.

 

No notable opaque spots or irregularities were observed on 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: 1.

As the mean IVIS was < 3, the test item was considered asa test chemical not requiring classification for eye irritation or serious eye damage (UN GHS No Category).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation study in rabbits (Blanchard 2003) :

A study was performed to assess the skin irritation potential of test material to the rabbit. The method followed was that described in OECD test guideline n°404.

Three rabbits were each administered a single dermal dose of 0.5 ml of the test substance an observed for up to 12 days. The effects of a 3 minutes, 60 minutes and 4 hours exposure were initially investigated using a screen animal.

Very slight dermal irritation was elicited following a single semi-occlusive application of test material to intact rabbit skin for 3 and 60 minutes.

A single semi-occlusive application of test material to intact rabbit skin for 4 hours alicited slight to well-defined dermal irritation in all animals.

The mean scores for these reactions at approximately 24, 48 and 72 hours after administration, calculated separately for each animal are : 1.3, 2 and 1 for erythema, and 0.3,1.3 and 1 for oedema.

Based on these results, the test material is considered to be slightly skin irritating.

 

In vitro eye irritation/corrosion study - BCOP (Gerbeix 2017):

 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 can identify chemicals inducing serious eye damage and chemicals not requiring classification for eye irritation or serious eye damage (OECD Guideline 437).

A single experiment was performed using three corneas for each treated series (test item, positive control and negative control). Before the treatment, a first opacity measurement was performed on each cornea using an opacitometer. The test item was applied undiluted in a single experiment using a treatment time of 10 minutes and using open-chamber method. Negative and positive controls were applied using the same treatment time and using the closed-chamber method. At the completion of the treatment period, all items were removed from the front opening of the anterior chamber and the epithelia were rinsed. The corneas were then incubated for 2 hours (± 10 minutes) at +32°C before a second opacity measurement was performed. After the second opacity measurement, the medium of the anterior chamber was removed and filled with a fluorescein solution. The holders were then incubated vertically for 90 minutes (± 5 minutes) at +32°C. 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.

 No notable opaque spots or irregularities were observed on 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: 1.

As the mean IVIS was < 3, the test item was considered as a test chemical not requiring classification for eye irritation or serious eye damage (UN GHS No Category).

Justification for classification or non-classification

Based on the available data, no classification for skin/eye irritation is required for the registered substance according to the Regulation EC N°1272/2008.