Registration Dossier

Administrative data

Description of key information

Evaluation in an in vivo acute dermal irritation/corrosion test in rabbits found the test substance to be corrosive after 3-minutes of exposure as full skin destruction developed over the course of two to three days.

An in vivo acute eye irritation/corrosion tests performed in rabbits the test substance elicited positive corneal, iridial and conjunctival responses in all eyes at 1 and 24h. Because of the severe nature of the responses and following a veterinary inspection the rabbits were humanely sacrificed following the 24h reading.

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:
From 2003-09-17 to 2003-10-01
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)
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: CEGAV, Saint mars d'Egrenne, France
- Age at study initiation: 2 to 4 months old
- Weight at study initiation: 2.7 kg
- Housing: individually in a polystyrene cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18±3°C
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 air changes per hour
- Photoperiod : 12 hour light/dark cycle

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
other: each animal served as its own control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied : 0.5 ml undiluted




Duration of treatment / exposure:
Left flank: 3 minutes.
Right flank: 4 hours.
Observation period:
1, 24, 48, 72 hrs after removal the patch and then daily until day 14.
Number of animals:
1 male rabbit
Details on study design:
TEST SITE
- Area of exposure: right flank (application for 4 hours) and left flank (application for 3 minutes)
- % coverage: 2.5x2.5 cm
- Type of wrap if used: semi-occlusive

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test item was wiped off by means of a dry cotton pad.
- Time after start of exposure: 3 minutes or 4 hours

SCORING SYSTEM: according to OECD guideline 404 (draize scoring system)




Irritation parameter:
erythema score
Remarks:
after a 3-minute exposure
Basis:
animal #1
Time point:
other: mean of the 24, 48 and 72-hour readings
Max. score:
4
Reversibility:
not reversible
Remarks on result:
other: The mean irritation score was not calculable because erythema was masked by a withish area at the 48 and 72-hour readings. The score of the 24-hour reading was 2.
Irritation parameter:
edema score
Remarks:
after a 3-minute exposure
Basis:
animal #1
Time point:
other: mean of the 24, 48 and 72-hour readings
Score:
2.3
Max. score:
4
Reversibility:
fully reversible within: 11 days
Irritation parameter:
erythema score
Remarks:
after a 4-hour exposure
Basis:
animal #1
Time point:
other: mean of the 24, 48 and 72-hour readings
Max. score:
4
Reversibility:
not reversible
Remarks on result:
other: The mean irritation score was not calculable because erythema was masked by a withish area at the 48 and 72-hour readings. The score of the 24-hour reading was 2.
Irritation parameter:
edema score
Remarks:
after 4-hour exposure
Basis:
animal #1
Time point:
other: mean of the 24, 48 and 72-hour readings
Score:
3
Max. score:
4
Reversibility:
fully reversible within: day 8
Irritant / corrosive response data:
- The mean scores over 24, 48 and 72 hours were not calculable for erythema (evaluation masked by the whitish area) whatever the exposure period.
- The mean scores over 24, 48 and 72 hours for oedema were 2.0 and 3.0 after a 3-minute and a 4-hour exposure period respectively.
The important cutaneous reactions observed after the 3-minute and 4-hour exposures were considered to be the consequence of a tissular destruction.
See freetext "remarks on results" for detailed descriptions of the test sites.

After a 3-minute exposure, a very slight then well-defined erythema (grade 1 then 2) was observed 1 hour and 24 hours after patch removal. From day 2 up to day 10, the evaluation of the erythema was masked by a whitish (days 2 and 3) or brownish (day 4) area, then by a crust (days 5 to 10). Then a well-defined erythema (grade 2) was noted from day 11 until the end of the observation period (day 14). A slight oedema (grade 2) was recorded 24 and 48 hours after patch removal. The oedema became more pronounced (grade 3) from day 3 up to day 6. The evaluation of the oedema was then masked by the crust from day 7 until day 10. Crusts and/or dryness of the skin were observed from day 5 and persisted at the end of the observation period (day 14).

After a 4-hour exposure, a well-defined erythema (grade 2) was noted 1 hour after patch removal. From day 1 up to day 7, the evaluation of the erythema was masked by a whitish (days 1 to 3) or brownish (day 4) area, then by a crust (days 5 to 7). A moderate to severe erythema (grade 3) was then observed on day 8, and a well-defined erythema (grade 2) persisted until the end of the observation period (day 14). A slight then moderate oedema (score 2 then 3), the evaluation of which was masked by a crust on day 7, was recorded 1 hour after patch removal and from day 1 up to day 5, respectively. Dryness of the skin was noted from day 4 and persisted at the end of the observation period (day 14).

Interpretation of results:
Category 1B (corrosive) based on GHS criteria
Remarks:
Migrated information
Conclusions:
Under the experimental conditions of this study and according to the criteria laid down in CLP (Reg 1272/2008/EC) and directive 67/548/EEC, the test material is classified Skin corrosive category 1B, H314 and Corrosive R35 respectively.
Executive summary:

The potential of the test item to induce skin irritation was assessed in rabbit according to the OECD guideline 404 in compliance with the principles of Good Laboratory Practice regulations.

As possible irritant effetcs were anticipated, the test item was first evaluated in a single animal. The duration of exposure was 3 minutes on one flank and 4 hours on the other flank. A single dose of 0.5 ml was applied to the right flank (application for 4 hours) or the left flank (application for 3 minutes) of the rabbit under a semi-occlusive dressing. Skin reactions were observed 1, 24, 48, 72 hours after removal of the dressing and then daily until day 14.The mean values of the scores for erythema and oedema were calculated when possible.

After a 3-minute exposure, a very slight then well-defined erythema (grade 1 then 2) was observed on days 1 and 2. From day 3 up to day 11, the evaluation of the erythema was masked by a whitish (days 3 and 4) or brownish (day 5) area, then by a crust (days 6 to 11). Then a well-defined erythema (grade 2) was noted from day 12 until the end of the observation period (day 15).

A slight or moderate oedema (grade 2 or 3) was recorded on days 2 and 3 and from day 4 up to day 7, respectively. The evaluation of the oedema was then masked by the crust from day 8 until day 11.

Crusts and/or dryness of the skin were observed from day 5 and persisted at the end of the observation period (day 15).

 

After a 4-hour exposure, a well-defined erythema (grade 2) was noted on day 1. From day 2 up to day 8, the evaluation of the erythema was masked by a whitish (days 2 to 4) or brownish (day 5) area, then by a crust (days 6 to 8). A moderate erythema (grade 3) was then observed on day 9, and a well-defined erythema (grade 2) persisted until the end of the observation period (day 15).

A slight then moderate oedema (score 2 then 3), the evaluation of which was masked by a crust on day 8, was recorded on day 1 and from day 2 up to day 6, respectively.

Dryness of the skin was noted from day 5 and persisted at the end of the observation period (day 15).

 

As the test item showed corrosive properties on this first animal, the study was considered complete and the test item was not evaluated on other animals.

Whatever the exposure duration, well defined erythema was noted along with slight to moderate oedema, but scoring was made difficult by the appearance of white/brown areas and crusting. Symptoms had not disappeared after 14 days. The mean scores over 24, 48 and 72 hours for the animal were not calculable for erythema (evaluation masked by the whitish area); for oedema, the mean scores over 24, 48 and 72 hours were 2.3 and 3.0 after the 3-minute and the 4-hour exposure period respectively.

The test substance was evaluated as causing severe burns after a 3 minute exposure on rabbit skin.

As the full skin destruction developed over the course of two to three days following the 3 minute exposure, it should be classified Cat. 1B for skin corrosion.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (corrosive)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study is not performed under GLP but there was inhouse QA. The study was performed similar to OECD 405 reporting was not done in full detai.CoA is not included.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
no
Remarks:
but inhouse quality assurance authentication was performed
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Cheshire Rabbit Farms Limited
- Age at study initiation: no data
- Weight at study initiation: 2.5-3.0 kg
- Housing: housed individually in cages with a grid floor beneath which was a peat moss filled tray.
- Diet (e.g. ad libitum): Ad libitum: Special Diet Services (S.D.S.) Rabbit Diet of satisfactory analysis
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 ± 2
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

IN-LIFE DATES: no data
Vehicle:
unchanged (no vehicle)
Controls:
other: each animal served as its own control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml
Duration of treatment / exposure:
24h after which the animals were sacrificed for humane reasons
Observation period (in vivo):
observations were done at 1 and 24h
Number of animals or in vitro replicates:
two males and one female
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing : eye not rinsed

SCORING SYSTEM:OECD: Acute Eye Irritation/Corrosion Test i n Rabbits Grades for Ocular Lesions
Cornea
No ulceration or opacity ........................................... 0
Scattered or diffuse areas of opacity ,
details of iris clearly visible .................... .... ........... I*
Easily discernible translucent areas of
opacity, details of iris slightly obscured ........................ 2*
Nacreous areas of opacity, no details of iris
visible, size of pupil barely discernible ......................... 3*
Complete corneal opacity, iris not discernible ...................... 4*
Ulceration, absence of a gross
patch of corneal epithelium ....................................... 4*

Iris
Normal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0
Markedly deepened folds, congestion, swelling,
moderate circumcorneal injection (any of these or
combination of any thereof), iris still reacting
to light (sluggish reaction i s positive) .......................... l*
No reaction to light, haemorrhage, gross
destruction (any or all of these) ................................. 2*

Conjunctivae
Redness (refers to pal pebral and bul bar
conjunctivae excluding cornea and iris) :
Vessels normal ...................................................... 0
Some vessels definitely injected ..................................... 1
Diffuse crimson red, individual vessels
not easily discernible ........................................... 2*
Diffuse beefy red . . . . . . . . . . . . . . . . . . . . .... . . . . . . . . . . . . . . . . . . . . . . . . . . . 3*

Chemosis
No swelling ........................................................ 0
Any swelling above normal (including nictitating membrane) ........... 1
Obvious swelling with partial eversion of lids ....................... 2*
Swelling with lids about half closed ........ . ....... . ............. ... 3*
Swelling with lids more than half closed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4*
Ulceration or necrosis of palpebral and bulbar
conjunctivae ornictitating membrane ............................... 4*

Discharge
Mild ................... ... ........ ................ ...............1
Moderate ........................................................2
Severe ............................................................ 3

* F i gures indicate the grades considered positive.

TOOL USED TO ASSESS SCORE: standard illumination.
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24 h
Score:
1
Max. score:
1
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24 h
Score:
1
Max. score:
1
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24 h
Score:
2
Max. score:
2
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24 h
Score:
4
Max. score:
4
Reversibility:
other: For humane reasons the animal was killed after 24h
Remarks on result:
other: Additionally: severe discharge
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24 h
Score:
2
Max. score:
2
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24 h
Score:
1
Max. score:
1
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24 h
Score:
2
Max. score:
2
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24 h
Score:
4
Max. score:
4
Reversibility:
other: For humane reasons the animal was killed after 24h
Remarks on result:
other: Additionally: severe discharge
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24 h
Score:
2
Max. score:
2
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24 h
Score:
1
Max. score:
1
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24 h
Score:
2
Max. score:
2
Reversibility:
other: For humane reasons the animal was killed after 24h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24 h
Score:
4
Max. score:
4
Reversibility:
other: For humane reasons the animal was killed after 24h
Remarks on result:
other: Additionally: severe discharge
Irritant / corrosive response data:
Cornea and Iris
It was not possible to assess corneal reactions at 1h in any animal because o f intense conjunctival swelling. At 24 h positive corneal reactions (score1 and 2) were noted in all animals. Positive iridial responses (score 1) were noted in all eyes at 1 and 24 h.

Conjunctival
Moderate conjunctival redness (score 2) and severe chemosis (score 3-4) were noted in all eyes at 1 and 24 h. Severe discharge (score 3) was noted
in all eyes at 1 and 24h.
Other effects:
All animals were sacrificed after the 24h reading on humane grounds.

Eye irritation/corrosion test in rabbits.

Rabbit No/Sex

1 hour

24 hours

1

2

3

1

2

3

Cornea

+

+

+

1

2

2

Iris

1

1

1

1

1

1

Conjunctivae:

 

 

 

 

 

 

   redness

2

1

2

2

2

2

   chemosis

4

3

4

4

4

4

   discharge

3

3

3

3

3

3

Total

10

8

10

11

12

12

+ = unable to read due to intense swelling

Interpretation of results:
corrosive
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
The test substance elicited positive corneal, iridial and conjunctival responses in all eyes at 1 and 24h. Because of the severe nature of the responses and following a veterinary inspection the rabbits were humanely sacrificed following the 24h reading.
Executive summary:

The potential of the test substance to induce eye irritation was assessed in 3 rabbits according to a method similar to OECD guideline 405.

A statement that the report was audited by the Quality Assurance Unit of the testing laboratory was provided.

A single dose of 0.1ml of the testsubstance was instilled into one eye, the other eye was not treated and served as control.The eyes were not rinsed after administration of the test item. Ocular reactions were observed 1 hour and 24 hours later.

It was not possible to assess corneal reactions at 1 hour in any animal because of intense conjunctival swelling. At 24 hours corneal scores of 1 or 2 were noted in all animals. Positive iridial scores of 1 were noted in all animals at 1 and 24 hours. Moderate conjunctival redness (score 2) and severe chemosis (score 4) were noted in all rabbits at 1 and 24 hours. Severe discharge was also noted in all rabbits.

Due to the severe nature of the reactions observed, the rabbits were sacrificed after the 24 hour recording on humane grounds. The authors concluded that the test substance was severely irritating to the eye.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion:

Two valid studies were recorded for this endpoint. The study of Ollivier (2003) was reliable without restrictions and the study of Cuthbert (1983) was reliable with some restrictions due to limited information reported about the tested substance and the lack of an observation period for the reversibility. The study of Ollivier (2003) was identified as the key study.

 

In the study of Ollivier (2003), the potential of the test item to induce skin irritation was assessed in rabbit according to the OECD guideline 404 in compliance with the principles of Good Laboratory Practice regulations.

As possible irritant effects were anticipated, the test item was first evaluated in a single animal. The duration of exposure was 3 minutes on one flank and 4 hours on the other flank. A single dose of 0.5 ml was applied to the right flank (application for 4 hours) or the left flank (application for 3 minutes) of the rabbit under a semi-occlusive dressing. Skin reactions were observed 1, 24, 48, 72 hours after removal the dressing and then daily until day 14. The mean values of the scores for erythema and oedema were calculated when possible.

As the test item showed corrosive properties on this first animal, the study was considered complete and the test item was not evaluated on other animals.

Whatever the exposure duration, well defined erythema was noted along with slight to moderate oedema, but scoring was made difficult by the appearance of white/brown areas and crusting, which was visible from day 2 following 4 hour exposure and from day 3 following 3 minute exposure. Symptoms had not disappeared after 14 days. The mean scores over 24, 48 and 72 hours for the animal were not calculable for erythema (evaluation masked by the whitish area); for oedema, the mean scores over 24, 48 and 72 hours were 2.3 and 3.0 after the 3-minute and the 4-hour exposure period respectively. Possibly the removal of the test substance by wiping with a dry cotton pad was not complete, leading to similar residues after both exposures. The test substance was evaluated as causing severe burns after a 3 minute exposure on rabbit skin.

 

The objective of the study of Cuthbert (1983b) was to evaluate the dermal irritation/ corrosion potential of the substance in rabbits according to a method similar to OECD guideline 404. A statement that the report was audited by the Quality Assurance Unit of the testing laboratory was provided. A single dose of 0.5g was applied to the skin of 3 rabbits under an occlusive dressing for a 4 -hour exposure period, after which the skin was wipe with damp tissues. Skin reactions were observed 1, 24, 48 and 72 hours after removal of the dressing but no further observation period was included in the study protocol. The same procedure was applied with the internal positive control, a 10% aqueous solution of lauryl sulphate. The mean values of the scores for erythema and oedema were calculated for each animal.

The mean scores over 24, 48 and 72 hours for individual animals were 2.0, 2.0 and 2.0 for erythema and 4.0, 4.0 and 4.0 for oedema. Responses had not diminished at the 72-hour reading. The positive control resulted in very slight to well defined erythema up to 48 hours and a very slight to slight oedema was noted up to 24 hours. All sites had returned to normal at 72 hours.

Based on these results, the authors concluded that the test substance was severely irritating to skin following a 4-hour occlusive exposure. A major deficiency is lack of observation of possible recovery after 72 hours at which time the maximum response was still observed.

 

Eye Irritation / corrosion:

One study is recorded for this endpoint. The study of Cuthbert (1983) was reliable with some restrictions mainly due to limited information reported about the tested substance.

 

In the study of Cuthbert (1983), the potential of the substance to induce eye irritation was assessed in 3 rabbits according to a method similar to OECD guideline 405. A statement that the report was audited by the Quality Assurance Unit of the testing laboratory was provided.

A single dose of 0.1ml of the test substance was instilled into one eye, the other eye was not treated and served as control. The eyes were not rinsed after administration of the test item. Ocular reactions were observed 1 hour and 24 hours later.

It was not possible to assess corneal reactions at 1 hour in any animal because of intense conjunctival swelling. At 24 hours corneal scores of 1 or 2 were noted in all animals. Positive iridial scores of 1 were noted in all animals at 1 and 24 hours. Moderate conjunctival redness (score 2) and severe chemosis (score 4) were noted in all rabbits at 1 and 24 hours. Severe discharge was also noted in all rabbits.

Due to the severe nature of the reactions observed, the rabbits were sacrificed after the 24 hour recording on humane grounds. Based on this, the results are interpreted as corrosive to the eyes.

Justification for selection of skin irritation / corrosion endpoint:

Study of highest quality and validity.

Justification for selection of eye irritation endpoint:

Only study available.

Effects on skin irritation/corrosion: corrosive

Effects on eye irritation: corrosive

Justification for classification or non-classification

Skin Irritation:

According to the results of the key study (Ollivier, 2003) and the criteria laid down in EU directive67/548/EEC, the substance is classified Corrosive with the risk phrase R35.

In this in vivo dermal corrosion study in rabbits, the effects indicative for corrosion were seen from the second day onward. Consequently, GHS classification 1B is appropriate, with hazard statement H314: Causes severe skin burns and eye damage.

 

Eye Irritation:

According to the results of the study carried out by Cuthbert (1983), the substance must be considered as highly irritating for eyes and warrants a classification in category 1 , eye damage according to CLP regulation (Reg 1272/2008/EC) and a classification severely irritating for eyes Xi, R41 according to the directive 67/ 548/ EEC.

Respiratory tract irritation: 

There is no information is available following exposure via inhalation. The substance is a viscous fluid with a very low vapourpressure of <3 10-7 Pa at 23 °C. Due to its physical state and physical chemical properties, it is unlikely that it will form inhalable dust, mist or fumes limiting the possible extend of exposure via inhalation. Inhalation of aerosols may cause respiratory irritation due to the corrosive properties of the substance.

For STOT-SE Cat 3 Respiratory Tract Irritation (RTI): Although the substance is very corrosive substances, the low vapour pressure prohibits occurrence of respiratory irritation by vapour. The classification of corrosive is already considered to implicitly cover the potential of RTI and additional Cat.3 is considered to be superfluous (Guidance CLP Ch. 3.8.2.5)