Registration Dossier

Administrative data

Description of key information

SKIN
Not corrosive (in vitro, Reconstructed Human Epidermis (RHE) model), OECD 431, EU Method B.40, Warren (2013a).
Not irritating (in vitro, Reconstructed Human Epidermis (RHE) model), OECD 439, EU Method B.46, Warren (2013b).
EYE
In vivo, Non-irritant (Rabbit, male), OECD 405, EU Method B.5, Sanders (2013b).
In vitro, Non-iritant, (in vitro, SkinEthic Reconstituted Human Corneal model), Warren (2013c).

Key value for chemical safety assessment

Skin irritation / corrosion

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
Remarks:
an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 February 2013 to 28 February 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed to standardised guidelines OECD 405 and EU Method B.5 and in line with GLP. The study was reported to a high standard, sufficient to assess the quality of the data presented.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Strain: Hsdlf:NZW
- Sex: male.
- Age at study initiation: 12 - 20 weeks.
- Weight at study initiation: 2.40 - 2.66 kg.
- Housing: individually in suspended cages.
- Diet: rabbit diet provided ad libitum.
- Water: mains drinking water, ad libitum.
- Acclimation period: at least 5 days.

Immediately before the start of the test, both eyes of the test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): at least 15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark.
Vehicle:
unchanged (no vehicle)
Controls:
other: The contra lateral eye was used as an untreated control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL (98 mg).
Duration of treatment / exposure:
- Dose administration: 0.1 mL of the test material was placed into the conjunctival sac of the right eye of one rabbit, by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were then held together for about 1 second immediately after treatment, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes. After consideration of the ocular responses and initial pain reaction in the first treated animal, a second animal was treated.
Observation period (in vivo):
Animals were observed up to 72 hours post administration.
Number of animals or in vitro replicates:
One animals initially, followed by a further animal once the irritation potential was fully assessed in the first animal.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: Irrigation was not performed.

SCORING SYSTEM:
The reactions observed 1, 24, 48 and 72 hours following treatment were scored in accordance with the criteria of Draize (1977) and assessed using a modified form of the Kay and Calandra system (1962). Any other ocular effects were also noted.

TOOL USED TO ASSESS SCORE:
Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

ADDITIONAL OBSERVATIONS
Any clinical signs of toxicity, if present, were noted.
Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Irritation parameter:
conjunctivae score
Remarks:
chemosis
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0.66
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritant / corrosive response data:
No corneal or iridial effects were noted during the study.
Minimal conjunctival irritation was noted in all treated eyes one hour after treatment and at the 24 and 48 hour observations.
Both treated eyes appeared normal at the 72 hour observation.

The test material produced a maximum group mean score of 3.0 and was classified as a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.
Other effects:
Both animals showed expected gain in body weight during the study.

Table 3: Ocular Irritation Results

Parameter 

Animal

Male 1

Male 2

Time after treatment (hours)

1  

24

48

72

Mean*

1  

24

48

72

Mean*

Cornea:

Degree of opacity

0

0

0

0

0

0

0

0

0

0

Area of cornea involved

0

0

0

0

0

0

0

0

0

0

Iris:

Iris

0

0

0

0

0

0

0

0

0

0

Conjunctivae:

Redness

1

1

1

0

0.66

1

1

1

0

0.66

Chemosis

1

0

0

0

0

0

0

0

0

0

Discharge

0

0

0

0

0

0

0

0

0

0

* Mean at 24, 48 and 72 hours.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the test, the test material only elicited slight reactions in any of the animals during the course of the study that meant the test material does not require classification as an eye irritant.
Executive summary:

The eye irritation potential of the test material was determined in a GLP study which was conducted in accordance with standardised guidelines OECD 405 and EU Method B.5. During the study, 0.1 mL of test material was placed into one eye of each of two rabbits and was assessed for up to 72 hours to determine the grade of ocular reaction. No corneal or iridial effects were noted during the study. Minimal conjunctival irritation was noted in all treated eyes one hour after treatment and at the 24 and 48 hour observations. Both treated eyes appeared normal at the 72 hour observation.

Under the conditions of the test, the test material only elicited slight reactions in any of the animals during the course of the study that meant the test material does not require classification as an eye irritant.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin

In the key study, Warren (2013a), the corrosivity potential of the test material was determined using the validated EPISKIN™ in vitro Reconstructed Human Epidermis (RHE) model. The study was performed in compliance with GLP and in accordance with the standardised guidelines OECD 431 and EU Method B.40 and as such, the study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality described in Klimisch (1997).

Under the conditions of the study the relative mean viability of the test item treated tissues were: 240 minutes exposure: 104.0 % ; 60 minutes exposure: 124.5 % ; 3 minutes exposure: 10.5 %. Since the relative mean viability was ≥ 35 % at 240 minutes, the test material was considered to be non-corrosive to skin.

 

Following on from the negative in vitro skin corrosion study, the skin irritation potential of the test material was determined in the key study Warren (2013b) using the validated EPISKIN™ in vitro Reconstructed Human Epidermis (RHE) model. The study was performed in compliance with GLP and in accordance with the standardised guidelines OECD 439 and EU Method B.46 and as such, the study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality described in Klimisch (1997).

The quality control criteria required for acceptance of results in the test were satisfied. Under the conditions of the study, the relative mean viability of the test material treated tissues was 92.7 % after the 15 minute exposure period, since this was > 50% the test material was concluded to be a non-irritant to the skin.

Eye

In the supporting study, reported by Warren (2013c) the in vitro eye irritation potential of the test material was determined in a non-standardised guideline study performed in line with good scientific practices and in line with GLP. The reliability of the study was assigned a reliability score of 2 in accordance with the principles for assessing data quality in accordance with Klimisch (1997) due to the pre-validation status of the protocol. The irritation potential was assessed using the SkinEthic Reconstituted Human Corneal model after a treatment period of 10 minutes.

Solution A served as the negative control and 2 % w/v Sodium Dodecyl Sulphate served as the positive control. Under the conditions of the study the relative mean viability of the test material treated tissues after a 10 minute exposure was 100.4 %. It was, therefore, considered unnecessary to proceed with tissue histopathology. Since the relative mean viability was ≥ 60 %, the test material was considered to be a non-irritant in accordance with the study protocol.

Following on from the negative in vitro eye irritation study, the in vivo eye irritation potential of the test material was determined in the key study, reported by Sanders (2013b). The study was performed in line with standardised guidelines (OECD 405 and EU Method B.5) and in accordance with GLP, the study was therefore assigned a reliability score of 1 in accordance with the principles for assessing data quality as described in Klimisch (1997). During the study, 0.1 mL of test material was placed into one eye of each of two rabbits and was assessed for up to 72 hours to determine the grade of ocular reaction.

No corneal or iridial effects were noted during the study. Minimal conjunctival irritation was noted in all treated eyes one hour after treatment and at the 24 and 48 hour observations. Both treated eyes appeared normal at the 72 hour observation.

Under the conditions of the test, the test material only elicited slight reactions in any of the animals during the course of the study that meant the test material does not require classification as an eye irritant.


Justification for selection of skin irritation / corrosion endpoint:
Both in vitro studies have been selected as key since they both address different endpoints, skin corrosivity and irritation. Both studies were performed under GLP conditions and in accordance with standardised guidelines. Studies were reported to a good standard and were assigned a reliability score of 1 in accordance with Klimisch (1997.)

Justification for selection of eye irritation endpoint:
The study reported by Sanders (2013b) was selected as the key study since it is an vivo study to determine eye irritation potential of test material. Supporting information is available in the form of an in-vitro study. The key study was performed to a high standard in accordance with GLP and standardised guidelines, and was assigned a reliability score of 1 in line with Klimisch (1997).

Justification for classification or non-classification

Skin

In accordance with the criteria for classification and labelling as defined in Regulation (EC) No. 1272/2008 (CLP) and Directive 67/548/EEC (DSD), the test material does not require classification for skin irritation and corrosion.

Eye

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008 and Directive 67/548/EEC (DSD), the test material does not require classification for eye irritation.