Registration Dossier

Administrative data

Description of key information

0.5 g of the test item was mixed to a paste in water before application to the skin.

RESULT: Very slight erythema with and without and very slight to slight oedema was observed in three intact and six abraded sites at the 24 hours reading and three abraded sites at the 72 hours reading.

The primary eye irritation potential of the test item was investigated according to a method compatible with OECD test guideline No. 405. The test item was applied by instillation of 0.1 mL into the right eye of each of two young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48, 72 hours and 7 and 14 days after test item instillation. 

 The mean score was calculated separately for each animal across three scoring times for corneal opacity, iritis, redness and chemosis of the conjunctivae. The individual mean scores for corneal opacity was 0.0 for both animals. The individual mean scores for iritis were 0.3 for both animals. The individual mean scores for the conjunctivae were 2.0 for reddening and 1.7 and 2.0 for chemosis. Small areas of petechial hemorrhage on the nictitating membrane were noted in one treated eye at the 48 and 72‑Hour observations.

 

The instillation of the test iteminto the eye resulted in conjunctival irritation. These effects were reversible in both animals. No abnormal findings were observed in the cornea of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.

  

According to the findings in this study, the test item meets the criteria for classification as ‘Irritating to Eyes’ (Category 2) according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

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:
1974
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Remarks:
Early study, but reporting the main relevant details, No GLP, short report
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
other: The Food and Drug Administration of the U.S .A. in The Federal Regi5ter (17 September 1964 § 191 .11)
GLP compliance:
no
Remarks:
pre-dates GLP
Species:
rabbit
Strain:
not specified
Type of coverage:
occlusive
Preparation of test site:
clipped
Vehicle:
water
Controls:
not required
Amount / concentration applied:
0,5 g of test substance was mixed to a paste in water before application
Duration of treatment / exposure:
24 h
Observation period:
72 hours
Number of animals:
6
Details on study design:
All animals were treated on intact skin (left) and on abraded skin (right)
Irritation parameter:
erythema score
Remarks:
intact skin
Basis:
mean
Time point:
other: 24 h
Score:
0.5
Max. score:
1
Reversibility:
fully reversible within: 72 h
Irritation parameter:
edema score
Remarks:
intact skin
Basis:
mean
Time point:
other: 24 h
Score:
0.166
Max. score:
1
Reversibility:
fully reversible within: 72 h
Irritant / corrosive response data:
Only 3 of 6 animals showed slight erythema on intact skin and 1 of these additionally a slight edema.
Other effects:
None

Animal

Effect

24 h

72 h

Intact

intact

1

Erythema

0

0

 

Edema

0

0

2

Erythema

0

0

 

Edema

0

0

3

Erythema

1

0

 

Edema

1

0

4

Erythema

0

0

 

Edema

0

0

5

Erythema

1

0

 

Edema

0

0

6

Erythema

1

0

 

Edema

0

0

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Findings were not sufficient to warrant classification under 1272/2008 CLP
Executive summary:

0.5 g of the test item was mixed to a paste in water before application to the skin.

RESULT

Very slight erythema with and without and very slight to slight oedema was observed in three intact and six abraded sites at the 24 hours reading and three abraded sites at the 72 hours reading.

Findings were not sufficient to warrant classification under 1272/2008 CLP

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation, other
Remarks:
in vivo study
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 02 June 215 and 07 July 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.
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
Qualifier:
according to
Guideline:
other: US EPA, OCSPP (OPPTS 870.2400, 1998)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No. 8147, amended 10 December 2002
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.59 or 3.20 kg and were 12 to 20 weeks old. After an acclimatization period of at least 5 days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
Vehicle:
unchanged (no vehicle)
Controls:
other: The left eye remained untreated and was used for control purposes.
Amount / concentration applied:
0.1 mL of the test item, which was found to weigh approximately 92 mg
Duration of treatment / exposure:
Up to 1 hour
Observation period (in vivo):
14 days
Number of animals or in vitro replicates:
2 animals were tested in total. (After consideration of the ocular responses produced in the first treated animal, an additional animal was treated. )
Details on study design:
Immediately before the start of the test, both eyes of the provisionally selected 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.

Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre dose anesthesia of ocular anesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.

A volume of 0.1 mL of the test item, which was found to weigh approximately 92 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid 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 left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1.

Eight hours after test item application, a subcutaneous injection of post dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.

After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated.

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation (Draize, J.H, 1977) given in Appendix 2.

Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Any clinical signs of toxicity, if present, were also recorded.

Additional observations were made on Days 7 and 14 to assess the reversibility of the ocular effects.

Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Irritation parameter:
cornea opacity score
Basis:
animal: 75082 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
cornea opacity score
Basis:
animal: 75111 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
iris score
Basis:
animal: 75082 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
0.3
Max. score:
2
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
iris score
Basis:
animal: 67747 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
0.3
Max. score:
2
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
other: redness
Basis:
animal: 75082 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: Small areas of petechial hemorrhage on the nictitating membrane
Irritation parameter:
other: redness
Basis:
animal: 75111 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
animal: 75082 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
animal: 75111 Male
Time point:
other: Mean Values after 24, 48 and 72 hours
Score:
1.7
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
Ocular Reactions

Individual and mean scores for ocular irritation are given in Table 1.

No corneal effects were noted during the study.

Iridial inflammation was noted in both treated eyes 1 and 24 hours after treatment.

Severe conjunctival irritation was noted in one treated eye and moderate conjunctival irritation was noted in the other treated eye 1 hour after treatment with moderate conjunctival irritation noted in both treated eyes at the 24 and 48 Hour observations. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye at the 72 Hour observation with minimal conjunctival irritation noted in both treated eyes at the 7 Day observation.

Small areas of petechial hemorrhage on the nictitating membrane were noted in one treated eye at the 48 and 72 Hour observations.

Both treated eyes appeared normal at the 14-Day observation.

No corrosive effects were noted during the study. The test item did not induce significant or irreversible damage to the rabbit eye.
Other effects:
Body weight

Individual body weights and body weight change are given in Table 2.

Both animals showed expected gain in body weight during the study.

Interpretation of Results

Data were summarized in tabular form, showing for each animal the irritation scores for the designated observation time, a description of the degree and nature of irritation, the presence of serious lesions and non-ocular effects. The scores of each animal at the following reading times (24, 48 and 72 hours) were used in calculating the respective mean values for each type of lesion.

 

The results were interpreted according to Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.

Table 1     Eye Irritation Scores

Individual Scores for Eye Irritation

Rabbit Number and Sex

IPR

Evaluation interval

Corneal Opacity

Area of Corneal Opacity

Iris

Conjunctivae

Redness

Chemosis

Discharge

75082Male

0

1 Hour

0

0

1

2

3

3

75111Male

0

0

0

1

2

3

2

75082Male

 

24 Hour

0

0

1

2

2

2

75111Male

 

0

0

1

2

2

2

75082Male

 

48 Hour

0

0

0

2Pt

2

2

75111Male

 

0

0

0

2

2

1

75082Male

 

72 Hour

0

0

0

2Pt

2

1

75111Male

 

0

0

0

2

1

0

75082Male

 

7 Days

0

0

0

1

1

0

75111Male

 

0

0

0

1

1

0

75082Male

 

14 Days

0

0

0

0

0

0

75111Male

 

0

0

0

0

0

0


IPR=Initial pain reaction

Pt =       Small areas of petechial hemorrhage on the nictitating membrane

Table 1 (continued)  Eye Irritation Scores

Mean Values after 24, 48 and 72 Hours

Rabbit Number
and Sex

Number of available data points

Corneal Opacity

Iris

Conjunctivae

Redness

Chemosis

75082Male

3

0.0

0.3

2.0

2.0

75111Male

3

0.0

0.3

2.0

1.7

 

 

Assessment According to Regulation (EC) No. 1272/2008

Evaluated Intervals

Corneal Opacity

Iris

Conjunctivae

Redness

Chemosis

24 Hours

Not classified

Not classified

Classified

Not classified

48 Hours

72 Hours

 

Table 2     Individual Body Weights and Body Weight Change

Rabbit Number
and Sex

Individual Body Weight (kg)

Body Weight Change (kg)

Day 0

Day 14

75082Male

2.59

2.81

0.22

75111Male

3.20

3.33

0.13

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the findings in this study, the test item meets the criteria for classification as ‘Irritating to Eyes’ (Category 2) according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.
Executive summary:

The primary eye irritation potential of the test item was investigated according to a method compatible with OECD test guideline No. 405 and Method B5. The test item was applied by instillation of 0.1 mL into the right eye of each of two young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48, 72 hours and 7 and 14 days after test item instillation. 

 

The mean score was calculated separately for each animal across three scoring times (24, 48 and 72 hours after instillation) for corneal opacity, iritis, redness and chemosis of the conjunctivae. The individual mean scores for corneal opacity was 0.0 for both animals. The individual mean scores for iritis were 0.3 for both animals. The individual mean scores for the conjunctivae were 2.0 for reddening and 1.7 and 2.0 for chemosis. Small areas of petechial hemorrhage on the nictitating membrane were noted in one treated eye at the 48 and 72‑Hour observations.

 

The instillation of the test item into the eye resulted in conjunctival irritation. These effects were reversible and were no longer evident 14 days after treatment in both animals (end of the observation period). No abnormal findings were observed in the cornea of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.

 

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

 

According to the findings in this study, the test item meets the criteria for classification as ‘Irritating to Eyes’ (Category 2) according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Irritant to eyes: H319 (Cat 2)

 

The instillation of the test item into the eye resulted in conjunctival irritation. These effects were reversible in both animals. No abnormal findings were observed in the cornea of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.