Registration Dossier

Administrative data

Description of key information

GLP compliant, in vivo acute dermal irritation/corrosion study in the rabbit, according to OECD 404.
GLP compliant, in vivo acute eye irritation/corrosion study in the rabbit, according to OECD 405.

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:
17 September 2003 to 08 December 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
ANIMALS AND ANIMAL HUSBANDRY
- Three New Zealand White rabbits were supplied by David Percival Ltd, Moston, Sandbach, Cheshire, UK.
- At the start of the study the animals were in the weight range 2.0 kg to 3.5 kg and were 12 to 20 weeks old.
- After an acclimatisation 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.
- Animals were individually housed in suspended metal cages.
- Free access to mains drinking water and food (Certified Rabbit Diet (Code 5322) supplied by International Product Supplies Limited, Wellingborough, Northants, 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.
- Temperature and relative humidity were set to achieve limits of 17 °C to 23 °C and 30 % to 70 % respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study.
- The rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.
- 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.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 mL
Duration of treatment / exposure:
Four hours
Observation period:
14 days
Number of animals:
Three
Details on study design:
PREPARATION OF TEST MATERIAL
- The absorption of the test item was not determined.

MEASUREMENT OF pH
- The pH of the test material was determined prior to commencement of the study and found to be 4.7 for a 10 % w/w aqueous preparation.
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 105 (male)
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 108 (male)
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 111 (male)
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
1.67
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 105 (male)
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
1.67
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 108 (male)
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 111 (male)
Irritant / corrosive response data:
- Individual scores for erythema/eschar and oedema are given in Table 1 (attached).
- Very slight to well-defined erythema was noted at all treated skin sites one hour after patch removal with well-defined erythema at all treated skin sites at the 24, 48 and 72 hour observations.
- Very slight to slight oedema was noted at all treated skin sites one hour after patch removal and at the 24 and 48 hour observations. Very slight oedema was noted at all treated skin sites at the 72 hour observation.
- Light brown discolouration of the epidermis was noted at all treated skin sites at the 24, 48 and 72 hour observations with loss of skin elasticity noted at all treated skin sites at the 72 hour observation.
- Crust formation, which prevented evaluation of erythema and oedema, was noted at all treated skin sites at the 7 day observation.
- All treated skin sites appeared normal at the 14 day observation.
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test material produced a primary irritation index of 3.3 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme. However, the test material is not classified under the terms of Regulation (EC) No 1272/2008 because the mean scores for three animals calculated using data from 24, 48 and 72 hour observations were < 2.3 for erythema and edema. No corrosive effects were noted.
Executive summary:

INTRODUCTION

The study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method was designed to meet the requirements of the OECD Guidelines for the Testing of Chemicals No 404 "Acute Dermal Irritation/Corrosion" (adopted 24 April 2002) and Commission Directive 92/69/EEC Method B.4 Acute Toxicity (Skin Irritation).

RESULTS

A single 4 hour semi-occluded application of the test material to the intact skin of three rabbits produced well-defined erythema and very slight oedema. Other reactions were light brown discolouration of the epidermis, loss of skin elasticity and crust formation, which prevented evaluation of erythema and oedema. All treated skin sites appeared normal at the 14 day observation.

CONCLUSION

The test material produced a primary irritation index of 3.3 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme. However, the test material is not classified under the terms of Regulation (EC) No 1272/2008 because the mean scores for three animals calculated using data from 24, 48 and 72 hour observations were < 2.3 for erythema and edema. No corrosive effects were noted.

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:
01 October 2003 to 08 December 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
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:
ANIMALS AND ANIMAL HUSBANDRY
- Three New Zealand White rabbits were supplied by David Percival Ltd, Moston, Sandbach, Cheshire, UK.
- At the start of the study the animals were in the weight range 2.0 kg to 3.5 kg and were 12 to 20 weeks old.
- After an acclimatisation 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.
- Animals were individually housed in suspended metal cages.
- Free access to mains drinking water and food (Certified Rabbit Diet (Code 5322) supplied by International Product Supplies Limited, Wellingborough, Northants, 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.
- Temperature and relative humidity were set to achieve limits of 17 °C to 23 °C and 30 % to 70 % respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study.
- The rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.
- 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:
no
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
Single application
Observation period (in vivo):
14 days
Number of animals or in vitro replicates:
Three
Details on study design:
PREPARATION OF TEST MATERIAL
- The absorption of the test material was not determined.

MEASUREMENT OF pH
- The pH of the test item was determined prior to commencement of the study and found to be 4.7 for a 10 % v/v aqueous preparation.

PROCEDURE
- 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 opthalmoscope.
- Only animals free of ocular damage were used.
- Initially, a single rabbit was treated. A volume of 0.1 mL of the test material 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 lids were held together for about 1 s immediately after treatment, to prevent loss of the test material, and then released.
- Immediately after adinistration of the test material, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1 (attached).
- After consideration of the ocular responses produced in the first treated animal, two additional animals were treated. In order to minimise pain on application of the test material, one drop of local anaesthetic (amethocaine hydrochloride 0.5 %, Chauvin Pharmaceuticals, Romford, Essex, UK) was instilled into both eyes of these animals 1 to 2 minutes before treatment.
- Assessment of ocular damage was made approximately 1 hour and 24, 48 and 72 hours following treatment according to the numerical evaluation (Draize scale) given in Appendix 2 (attached).
- Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard opthalmoscope.
- Additional observations were made on Days 7 and 14 to assess the reversibility of the ocular effects.

INTERPRETATION OF RESULTS
- Numerical scores corresponding to each animal, tissue and observation time were recorded.
- Data relating to the conjuntivae were designated by the letters A (redness), B (chemosis) and C (discharge).
- Data relating to the iris were designated by the letter D.
- Data relating to the cornea were designated by the letter E (degree of opacity) and F (area of cornea involved).
- Score for conjuntivae = (A + B + C) * 2
- Score for iris = D * 5
- Score for cornea = (E x F) * 5
- Using the numerical data obtained, a modified version of the system described by Kay and Calandra (1962) was used to classify the ocular irritancy potential of the test material (see Appendix 3, attached). This was achieved by adding together the scores for the cornea, iris and conjuntivae for each time point for each rabbit.
- The group means of the total scores for each observation were calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled classification of the eye irritancy potential of the test material.
- If evidence of irreversible ocular damage was noted, the test material would be classified as corrosive to the eye.

Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
animal #1
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: 178 male
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 48 h
Remarks on result:
other: 178 male
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 h
Remarks on result:
other: 178 male
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other: 178 male
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
animal #2
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 7 d
Remarks on result:
other: 14 male
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
0.67
Max. score:
2
Reversibility:
fully reversible within: 7 d
Remarks on result:
other: 14 male
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #2
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 14 male
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 14 male
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
animal #3
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
1.67
Max. score:
4
Reversibility:
fully reversible within: 7 d
Remarks on result:
other: 15 male
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 7 d
Remarks on result:
other: 15 male
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #3
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 15 male
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
mean score
Time point:
other: 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 d
Remarks on result:
other: 15 male

RESULTS

- Individual and group mean scores for ocular irritiation are given in Table 1 and Table 2 (attached).

- Scattered or diffuse corneal opacity was noted in one treated eye at the 24 hour observation with translucent corneal opacity at the 48 hour and 72 hour observation. Scattered or diffuse corneal opacity was alo noted in one other treated eye at the 48 hour and 72 hour observation.

- Iridial inflammation was noted in two treated eyes at the 24 hour observation. Iridial inflammation persisted in one treated eye, developed in one other treated eye at the 48 hour observation and persisted in these two treated eyes at the 72 hour observation.

- Moderate conjunctival irritation was noted in all treated eyes at 1 hour and 24 hours after treatment. Moderate conjunctival irritation was noted in two treated eyes with minimal conjunctival irritation noted in the remaining treated eye at the 48 hour observation. Moderate conjunctival irritation was noted in two treated eyes at the 72 hour observation with minimal conjunctival irritation at the 7 day observation.

- Pale appearance of the nictitating membrane was noted in one treated eye at the 48 hour and 72 hour observations.

- One treated eye appeared normal at the 72 hour observation and the remaining treated eyes appeared normal at the 14 day observation.

Interpretation of results:
other: Irritating to eyes (category 2)
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
The test material produced a maximum group mean score of 24.3 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. Since the mean scores for conjunctival redness and chemosis calculated from observations at 24, 48 and 72 hours were ≥ 2 in two out of three animals, classification as irritating to eyes (category 2) is required in accordance with Regulation (EC) No 1272/2008..
Executive summary:

INTRODUCTION

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of OECD Guidelines for the Testing of Chemicals No 405 "Acute Eye Irritation/Corrosion" (adopted 24 April 2002) and Commission Directive 92/69/EEC Method B.5 Acute Toxicity (Eye Irritation).

RESULT

A single application of the test material to the non-irrigated eye of three rabbits produced scattered or diffuse to translucent corneal opacity, iridial inflammation and moderate conjunctival irritation. Pale appearance of the nictitating membrane was also noted in one treated eye. One treated eye appeared normal at the 72 hour observation and the remaining treated eyes appeared normal at the 14 day observation.

CONCLUSION

The test material produced a maximum group mean score of 24.3 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. Since the mean scores for conjunctival redness and chemosis calculated from observations at 24, 48 and 72 hours were ≥ 2 in two out of three animals, classification as irritating to eyes (category 2) is required in accordance with Regulation (EC) No 1272/2008.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation

INTRODUCTION

The study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method was designed to meet the requirements of the OECD Guidelines for the Testing of Chemicals No 404 "Acute Dermal Irritation/Corrosion" (adopted 24 April 2002) and Commission Directive 92/69/EEC Method B.4 Acute Toxicity (Skin Irritation).

RESULTS

A single 4 hour semi-occluded application of the test material to the intact skin of three rabbits produced well-defined erythema and very slight oedema. Other reactions were light brown discolouration of the epidermis, loss of skin elasticity and crust formation, which prevented evaluation of erythema and oedema. All treated skin sites appeared normal at the 14 day observation.

CONCLUSION

The test material produced a primary irritation index of 3.3 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme. However, the test material is not classified under the terms of Regulation (EC) No 1272/2008 because the mean scores for three animals calculated using data from 24, 48 and 72 hour observations were < 2.3 for erythema and edema. No corrosive effects were noted.

Eye Irritation

INTRODUCTION

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of OECD Guidelines for the Testing of Chemicals No 405 "Acute Eye Irritation/Corrosion" (adopted 24 April 2002) and Commission Directive 92/69/EEC Method B.5 Acute Toxicity (Eye Irritation).

RESULT

A single application of the test material to the non-irrigated eye of three rabbits produced scattered or diffuse to translucent corneal opacity, iridial inflammation and moderate conjunctival irritation. Pale appearance of the nictitating membrane was also noted in one treated eye. One treated eye appeared normal at the 72 hour observation and the remaining treated eyes appeared normal at the 14 day observation.

CONCLUSION

The test material produced a maximum group mean score of 24.3 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.Since the mean scores for conjunctival redness and chemosis calculated from observations at 24, 48 and 72 hours were ≥ 2 in two out of three animals, classification as irritating to eyes (category 2) is required in accordance with Regulation (EC) No 1272/2008.


Effects on skin irritation/corrosion: irritating

Effects on eye irritation: irritating

Justification for classification or non-classification

Skin

The test material is not classified under the terms of Regulation (EC) No 1272/2008 because the mean scores for three animals calculated using data from 24, 48 and 72 hour observations were below the stated limits for erythema and edema and all treated sites appeared normal at the 14 day observation.

Eye

The mean scores for conjunctival redness and chemosis calculated from observations at 24, 48 and 72 hours were ≥ 2 in two out of three animals. Classification as irritating to eyes (category 2) is therefore required in accordance with Regulation (EC) No 1272/2008.