Registration Dossier

Administrative data

Description of key information

Skin irritation: not irritating, OECD 404, Moreno 1979
Skin irritation: not irritating, HRIPT, Wilson 1979
Eye irritation: not irritating, OECD 405, Teunissen 2013

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 vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP.
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
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: at least 6 weeks old
- Weight at study initiation: at least 1.0kg
- Housing: individually housed in labeled cages with perforated floors (dimensions 67 x 62 x 55 cm) and shelters.
- Diet (e.g. ad libitum): certified pellet diet (100g a day) and wooden sticks
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 40 - 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hour light/12 hour dark

IN-LIFE DATES: From: 04/02/13 To: 18/02/13
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): undiluted

Duration of treatment / exposure:
Each animal was treated by instillation of 0.1 mL of the test substance, in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control. Immediately after the 24-hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage.
Observation period (in vivo):
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 days after instillation of the test substance.
Number of animals or in vitro replicates:
3 males
Details on study design:
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 1 week s later, after considering the degree of eye irritation observed in the first animal.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none

SCORING SYSTEM:
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal luster) 0
Scattered or diffuse areas of opacity, details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacreous area, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity 4

Area of cornea involved:
No ulceration or opacity 0
One quarter or less but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4

IRIS
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia,
or injection, any of these or combination thereof, iris still reacting to light
(sluggish reaction is positive) 1
No reaction to light, hemorrhage, gross destruction (any or all of these) 2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal 0
Some blood vessels definitely hyperaemic (injected) 1
Diffuse, crimson color, individual vessels not easily discernible 2
Diffuse beefy red 3

Chemosis (refers to lids and/or nictitating membranes):
No swelling 0
Any swelling above normal (includes nictitating membranes) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals) 0
Any amount different from normal and/or lacrimation 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs (considerable area around the eye) 3

TOOL USED TO ASSESS SCORE: ophthalmic examination lamp
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
1.13
Max. score:
3
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
0.53
Max. score:
4
Reversibility:
fully reversible
Irritant / corrosive response data:
Irritation of the conjunctivae, which consisted of redness, chemosis and discharge was observed in all test animals. The irritation had completely resolved within 48 hours in one animal and within 7 days in the other two animals. No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage. There was no evidence of ocular corrosion.
Other effects:
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1. Individual scores

Animal Time after dosing Cornea Iris Conjunctivae
Opacity Area Fluorescein area Redness Chemosis Discharge
1 1h 0 0 - 0 2 2 2
24h 0 0 0 0 2 2 1
38h 0 0 - 0 2 1 1
72h 0 0 - 0 1 0 0
7d 0 0 - 0 0 0 0
2 1h 0 0 - 0 2 2 2
24h 0 0 0 0 2 1 1
38h 0 0 - 0 2 0 0
72h 0 0 - 0 1 0 0
7d 0 0 - 0 0 0 0
3 1h 0 0 - 0 2 2 2
24h 0 0 0 0 1 1 1
38h 0 0 - 0 1 0 0
72h 0 0 - 0 0 0 0
7d 0 0 - 0 0 0 0
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the test material is not irritating to New Zealand white rabbit eyes.
Executive summary:

The study was performed to assess the irritancy potential of the test material to the eye following a single application in the New Zealand White rabbit. The study was performed to GLP and the method was designed to meet the requirements of OECD Guidelines for the Testing of Chemicals No. 405. A volume of 0.1 ml of the test material was placed into the conjunctival sac of one eye of 3 animals. The other eye remained untreated and was used for control purposes. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. No corneal (opacity) or iridial effects were noted during the study. Minimal to moderate (grade 1 to 2) conjunctival irritation was noted for all animals at the 1 to 48 -hour observations.The irritation had completely resolved by 7 days in all animals. Under the conditions of this study the test material is not considered to be irritating to New Zealand White rabbit eyes.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

OECD 404 1979 - In an in vivo animal study the irritancy potential of the test material to the skin of the New Zealand White rabbit was assessed. The study was performed pre-GLP and followed a method similar to OECD 404 guideline. The test substance was evaluated in 6 New Zealand white rabbits. A dose of 0.5 ml test substance (undiluted), was applied to intact and abraded clipped dorsal skin site under a occlusive dressing for 24 hours. Skin observations were made 24 and 72 hours after patch removal. Very slight to well defined erythema was noted at 24 hours. Very slight edema was also noted at 24 hours. By 72 hours, all dermal scores were essentially zero. The test material produced a mean primary irritation index (PII) of 1.13 and is considered to be non-irritating to skin.

HRIPT 1979 - A study was performed to assess the irritancy potential of the test material in human volunteers. The study was performed following a modification of Draize Repeated Insult Patch Test method. The test substance was evaluated in 50 human volunteers. A dose of 0.2 g test substance, was applied to the upper arm of each subject under a semi-occlusive dressing for 24 hours. This was repeated 3 times a week for 3 weeks for a total of 9 applications. No skin effects were observed in any test subject. Under the conditions of this test, the test material is considered to be non-irritating to skin.

Eye irritation:

BCOP 2013 - A BCOP study was performed to assess the irritancy potential of the test material to the eye following exposure to bovine corneas. The study was performed to GLP and the method was designed to meet the requirements of OECD 437 guideline. A total of 3 corneas per treatment group were used. A volume of 750 microlitres of the test material was placed the cornea. The negative control group received saline and the positive control group received 10% benzalkonium chloride. For each group the corneas were incubated for 10 minutes at 32 degrees C. After the incubation the solutions were removed and the corneas were washed with MEM with phenol red (Eagle’s Minimum Essential Medium) and thereafter with cMEM. Possible pH effects of the test substance on the corneas were recorded. After the completion of the incubation period opacity determination was performed. Each cornea was inspected visually for dissimilar opacity patterns. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (10% (w/v) Benzalkonium Chloride) was 136 and was within the historical positive control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. The test material did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 11.2 after 10 minutes of treatment. Based on these results the test material is considered to be non irritating or corrosive in the Bovine Corneal Opacity and Permeability test.

OECD 405 2013 - The study was performed to assess the irritancy potential of the test material to the eye following a single application in the New Zealand White rabbit. The study was performed to GLP and the method was designed to meet the requirements of OECD Guidelines for the Testing of Chemicals No. 405. A volume of 0.1 ml of the test material was placed into the conjunctival sac of one eye of 3 animals. The other eye remained untreated and was used for control purposes. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. No corneal (opacity) or iridial effects were noted during the study. Minimal to moderate (grade 1 to 2) conjunctival irritation was noted for all animals at the 1 to 48 -hour observations.The irritation had completely resolved by 7 days in all animals. Under the conditions of this study the test material is not considered to be irritating to New Zealand White rabbit eyes.


Justification for selection of skin irritation / corrosion endpoint:
one in vivo klimisch 4 and one HRIPT klimisch 4 studies. No study was selected since all studies indicate with clear weight of evidence no adverse effects.

Justification for selection of eye irritation endpoint:
Four studies available; one in vivo klimisch 1, one in vitro klimisch 1 and two disregarded in vivo klimisch 4 studies. Study selected has the highest reliability supported by an in vitro study both indicating no adverse effects.

Justification for classification or non-classification

The substance does not meet classification criteria under EU Directive 67/548/EEC for dermal irritation.

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for dermal irritation.

 

The substance does not meet classification criteria under EU Directive 67/548/EEC for eye irritation.

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for eye irritation.