Registration Dossier

Administrative data

Description of key information

Skin: A single 24-Hour, occluded application of the test item to the intact skin of three rabbits produced a very slight erythema response in two animals at the 24-hour observation which had fully reversed by the 72-hour observation. The substance is not a skin irritant in rabbits.
Eye: A single application of the test item to the non-irrigated eye of three rabbits produced minimal conjunctival irritation. All treated eyes appeared normal at the 48-Hour observation. The test material is not an irritant in the eye of rabbits.

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:
14th March 1977 to 4th April 1977
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Older in vivo study used as read-across. No further in vivo studies conducted due to cosmetic use. See attached read across justification.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
No observation at 48 hours, as only mild irritation scores were observed it is not thought to impact the results of the test.
Principles of method if other than guideline:
The intact and abraded skin of 3 rabbits was used. The hair was clipped from the backs of the rabbits using angora clippers. Four areas of the back, places ca. 10 cm apart were designated for the patches. Areas 2and 3 were abraded by making four epidermal incisions (two perpendicular to two others in the area of the patch). The patches consisted of layers of light gauze cut in aquares (2.5 cm sides). The patches were secured to the area by thin bands of adhesive tape. The material was introduced (0.5 mL) beneath the patch. The entire trunks of the animals were then wrapped in clear plastic trunk bands. The test material and control, if using, were applied so that there were two applications to each of 3 animals (one intact, one abraded). The animals were immobilised for the 24 hour exposure period. The evaluations were made at 24 and 72 hours after application.
GLP compliance:
no
Remarks:
Prior to GLP
Specific details on test material used for the study:
Date
Species:
rabbit
Strain:
not specified
Details on test animals and environmental conditions:
No details on test animals and environment provided
Type of coverage:
occlusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 mL
Duration of treatment / exposure:
24 hour occlusive exposure. The sites were not washed after the patches were removed.
Observation period:
24 and 72 hours
Number of animals:
3
Details on study design:
The intact and abraded skin of 3 rabbits was used. The hair was clipped from the backs of the rabbits using angora clippers. Four areas of the back, places ca. 10 cm apart were designated for the patches. Areas 2and 3 were abraded by making four epidermal incisions (two perpendicular to two others in the area of the patch). The patches consisted of layers of light gauze cut in aquares (2.5 cm sides). The patches were secured to the area by thin bands of adhesive tape. The material was introduced (0.5 mL) beneath the patch. The entire trunks of the animals were then wrapped in clear plastic trunk bands. The test material and control, if using, were applied so that there were two applications to each of 3 animals (one intact, one abraded). The animals were immobilised for the 24 hour exposure period. The evaluations were made at 24 and 72 hours after application.

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
probability of weak irritation
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
probability of weak irritation
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
Intact skin only
The test material elicited a very slight erythema response in two animals at the 24-hour observation which had fully reversed by the 72-hour observation. No other irritation responses were noted.

Irritation effects on intact skin only are considered for classification purposes.

Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not a skin irritant in rabbits.
Executive summary:

Introduction

The study was performed in accordance with the US Federal Hazardous Substances Labelling Act to assess the irritancy potential of the test item to the skin of rabbits.

Method and materials

The intact and abraded skin of 3 rabbits was used. The hair was clipped from the backs of the rabbits using angora clippers. Four areas of the back, places ca. 10 cm apart were designated for the patches. Areas 2and 3 were abraded by making four epidermal incisions (two perpendicular to two others in the area of the patch). The patches consisted of layers of light gauze cut in aquares (2.5 cm sides). The patches were secured to the area by thin bands of adhesive tape. The material was introduced (0.5 mL) beneath the patch. The entire trunks of the animals were then wrapped in clear plastic trunk bands. The test material and control, if using, were applied so that there were two applications to each of 3 animals (one intact, one abraded). The animals were immobilised for the 24 hour exposure period. The evaluations were made at 24 and 72 hours after application and scored according to the Draize scheme.

Results

A single 24-Hour, occluded application of the test item to the intact skin of three rabbits produced a very slight erythema response in two animals at the 24-hour observation which had fully reversed by the 72-hour observation. No other irritation responses were noted.

Conclusion

The substance is not a skin irritant in rabbits.

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:
1977-03-14 to 1977-04-04
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Justification for type of information:
See attached justification
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
not applicable
Principles of method if other than guideline:
One tenth of a milliliter of the test material was instilled into the conjunctival sac of 3 albino rabbits. The test material was allowed to fall on the everted lower lid of each animal and the upper and lower lids were then gently held together for one second to prevent loss of the test material. All treated eyes remained unwashed.
The evaluations were performed at 1, 24, etc hours for a period of 7 days. A 2% instillation of fluorescin was made on each animal at least once during the course of the experimental period.
GLP compliance:
no
Remarks:
Prior to GLP
Species:
rabbit
Strain:
not specified
Details on test animals or tissues and environmental conditions:
No details on test animals and environment provided
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
7 days - eye unwashed.
Observation period (in vivo):
1 hour, 24, 48, 72 hours and then up to 7 days exposure.
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: hand-slit lamp / fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.67
Max. score:
20
Reversibility:
fully reversible within: 72 hours
Remarks on result:
positive indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
positive indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2.67
Max. score:
20
Reversibility:
fully reversible within: 72 hours
Remarks on result:
positive indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.67
Max. score:
20
Reversibility:
fully reversible within: 72 hours
Remarks on result:
positive indication of irritation
Irritant / corrosive response data:
Redness: All three animals exhibited conjunctival redness with a score of 2 on the Draize scale 1 hour after test material instillation. By the 24 hour observation the score was reduced to 1 for all animals with effects fully reversed by the 48-hour observation.
Minor conjunctival discharge was observed in 2 animals at the 1 hour observation, this was fully reversed in one animal by the 24 hour observation and in the remaining animal at the 48-hour observation.
One animal exhibited minor conjunctival chemosis at the 1-hour observation. This was fully reversed by the 24-hour observation. No other effects on eyes were noted during the course of the study.
Other effects:
No data
Interpretation of results:
GHS criteria not met
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material is not an irritant to the eyes of rabbit under the conditions of the study. The test item is not classifiable under CLP.
Executive summary:

Introduction

The study was performed in accordance with the US Federal Hazardous Substances Labelling Act.

Method

A volume of 0.1 mL of the test item was placed into the conjunctival sac of one eye of three rabbits, formed by gently pulling the lower li d away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The eyes were left unwashed and observations were perfomed one hour after installation and then daily for a period of 7 days. Assessment of ocular damage/irritation was made according to the numerical evaluation (Draize, J.H, 1977).

Results

A single application of the test item to the non-irrigated eye of three rabbits produced minimal conjunctival irritation. All treated eyes appeared normal at the 48-Hour observation.

Conclusion

The test material is not an irritant in the eye of rabbits. The test item is not classifiable under CLP.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The justification for read-across to the substance EC 255 -485 -3 is attached in Section 13 of the IUCLID.

Skin

The key study on the read-across substance EC 255 -485 -3 was performed in accordance with the US Federal Hazardous Substances Labelling Act to assess the irritancy potential of the test item to the skin of rabbits.

The intact and abraded skin of 3 rabbits was used. The hair was clipped from the backs of the rabbits using angora clippers. Four areas of the back, places ca. 10 cm apart were designated for the patches. Areas 2and 3 were abraded by making four epidermal incisions (two perpendicular to two others in the area of the patch). The patches consisted of layers of light gauze cut in aquares (2.5 cm sides). The patches were secured to the area by thin bands of adhesive tape. The material was introduced (0.5 mL) beneath the patch. The entire trunks of the animals were then wrapped in clear plastic trunk bands. The test material and control, if using, were applied so that there were two applications to each of 3 animals (one intact, one abraded). The animals were immobilised for the 24 hour exposure period. The evaluations were made at 24 and 72 hours after application and scored according to the Draize scheme.

A single 24-Hour, occluded application of the test item to the intact skin of three rabbits produced a very slight erythema response in two animals at the 24-hour observation which had fully reversed by the 72-hour observation. No other irritation responses were noted.

The substance is not a skin irritant in rabbits. In accordance with EU CLP Regulation (EC) No. 1272/2008 classification of this substance for skin irritation is not required.

Eye

in vitro

The key in vitro study on the substance was performed using the the MatTek Corporation EpiOcular Tissue Model in vitro toxicity testing system.

After the appropriate tissue preparation, 100 µL of the test article (10% in corn oil) and negative control (distilled water) were added to the millicells containing the EpiOcular samples. The six well plates containing the dosed EpiOcular samples were then incubated at 37°C, 5% CO2 and >= 90% humidity.

After the appropriate exposure period, each insert was individually removed from its plate and rinsed with phosphate buffered saline (PBS) to remove any residual material. Each was then rinsed for a second and third time. Following the three rinses, each Millicell was submerged in 5 mL of assay media for 10 mins at room temperature. This final soak removed any residual absorbed article. After the 10 mins, excess liquid was shaken off and each EpiOcular tissue was placed into 300 µL of MTT solution. The EpiOcular samples were then returned to the incubator.

After the 3 hour MTT exposure, each insert was removed and gently rinsed with PBS to remove any residual MTT solution. Excess PBS was shaken from each of the inserts, which were then blotted on the bottom on paper towels. The inserts were then placed into one well of a 24 well extraction plate. Each insert was then immersed in 2 mL of extration at room temperature overnight. After the extraction procedure,. the liquid within each insert was decanted back intothe well from which it was taken. The remaining extractant solution was then agitated and a 200 µL of each extract was removed for evaluation. A Dynatech MR 4000 Automatic Microplate Reader was used to determine the absorbance of each extract at 570 nm. With the absorbance of a negative control (distilled water) defined as 100%, the percentage absorbancies of the test and reference articles were determined.

The test material (10% in corn oil) elicited the following results

Exposure time              % inhibition              %Viability

5 min                             106                            -6

1 hour                            102                            -2

4 hours                           100                            0

The resulting ET50 would be greater than 256 minutes. The estimated Draize score is 0.

Under the conditions of the test, the test substance (10% in corn oil) is not irritating in vitro.

in vivo

The key in vivo study on the read-across substance EC 255 -485 -3 was performed in accordance with the US Federal Hazardous Substances Labelling Act.

A volume of 0.1 mL of the test item was placed into the conjunctival sac of one eye of three rabbits, formed by gently pulling the lower li d away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The eyes were left unwashed and observations were perfomed one hour after installation and then daily for a period of 7 days. Assessment of ocular damage/irritation was made according to the numerical evaluation (Draize, J.H, 1977).

A single application of the test item to the non-irrigated eye of three rabbits produced minimal conjunctival irritation. All treated eyes appeared normal at the 48-Hour observation.

The test material is not an irritant in the eye of rabbits. The test item is not classifiable under CLP.

Justification for classification or non-classification

Based on the read-across to EC 255 -485 -3:

Skin: The test material is not an irritant to the skin of rabbits

Eye: The test material is not an irritant in the eye of rabbits

In accordance with EU CLP Regulation (EC) No. 1272/2008 classification of this substance for irritation is not required.