Registration Dossier

Administrative data

Description of key information

Not skin irritating

Eye Dam. 1 (H318), according to the CLP Regulation (EC) No 1272/2008

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:
From August 09 to September 06, 1994
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted 17.07.1992
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: Interfauna UK Ltd Wyton, Huntingdon, England
- Age at study initiation: adult
- Weight at study initiation: the animals were weighed immediately before application of the test substance.
- Housing: the rabbits were individually housed in stainless steel cages with flat rod bases or plastica cages with perforated bases, under standardised conventional conditions.
- Diet: standard diet "Ssniff K4" approx. 100-120g per animal/day; once a day in the morning.
- Water: tap water ad libitum.
- Acclimation period: prior to use the animals were acclimated for approx. one week in the animal room.
- Health check: approx. 24 hours before the test fur was shaved from the dorso-lateral area of the trunk (6×6 cm) of each of the rabbits. Care was taken to avoid abrading the skin. Only animals with healty and intact skin were used.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 1.5 °C
- Humidity: 60-80 %
- Air changes:12-15 times per hour
- Photoperiod: from 6 am to 6 pm, 12 hours artifical illumination.
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
500 mg
Duration of treatment / exposure:
4 hours
Observation period:
14 days
Number of animals:
3 rabbits
Details on study design:
TEST SITE
- Area of exposure: test substance was moistened with deionised water and subsequently applied to a hypoallergenic Hansamed patch. A further patch was moistened with water.
- Type of wrap if used: the patches prepared in this way were placed in the opposite dorso-lateral areas of the trunk of each animal and were held in place with semiocclusive dressing for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing: after exposure time the dressing was removed and the exposed skin areas were carefully washed with water without altering the existing response or the integrity of the epidermis.
- Time after start of exposure: 4 hours.

SCORING SYSTEM
For each animal the Draize scores recorded at 24, 48 and 72 h were added up. The total was divided by 3 to give the irritation index. This index was separately calculated for erythema/eschar formation and for oedema formation.

Erythema and eschar formation
No erythema 0
Very slight erythema (barely perceptible) 1
WelI-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

Oedema formation
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimetre) 3
Severe oedema (raised more than I millimetre and extending beyond the area of exposure) 4

Interpretation of results - irritation index:
0.0 - 0.99 no irritation
1.0 - 1.99 slight irritation
2.0 - 2.99 moerate irritation
3.0 - 4.0 severe irritation
Irritation parameter:
edema score
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 2.3
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal: 3/3
Time point:
24/48/72 h
Remarks on result:
not determinable
Irritant / corrosive response data:
Evaluation of erythema was not possible in all three animals due to the intense colouration by test substance. Neverthless no other inflammatory signs (oedema or eschar formation) became apparent within the observation period of 7 days. Signs of toxicity were not observed. This evidence indicates no hazard potential to the skin and test substance may therefore be regarded as "not irritating" to the skin.

Individual reactions

Animal Reactions Score at
1 hr 24 hrs 48 hrs 72 hrs 7 d 14 d
D9 Erythema a a a a a -
D13 Erythema a a a a a -
B7 Erythema a a a a a -
D9 Oedema 0 0 0 0 0 -
D13 Oedema 0 0 0 0 0 -
B7 Oedema 0 0 0 0 0 -

-: not examined

a: exposed skin areas stained in colour of the test substance, evaluation of erythema was not possible.

Interpretation of results:
other: not irritating
Conclusions:
Not irritating.
Executive summary:

The irritant/corrosive potential of test substance was studied on the skin of rabbit in accordance with OECD guideline 404.

A volume of 0.5 g of test material was moistened with deionised water and subsequently applied to a hypoallergenic Hansamed patch. A further patch was moistened with water. The patches were held in place with semiocclusive dressing for the duration of the exposure period. After the exposure time of 4 hours, the dressing was removed and the exposed skin areas were carefully washed with water without altering the existing response or the integrity of the epidermis.

Evaluation of erythema was not possible in all three animals due to the intense colouration by test substance. However, no other inflammatory signs (oedema or eschar formation) became apparent within the observation period of 7 days. Signs of toxicity were not observed. This evidence indicates no hazard potential to the skin and the test substance may therefore be regarded as "not irritating" to the skin.

Conclusion

Not skin irritating.

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:
From August 09th to September 06th, 1994
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted 24.02.1987
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Interfauna UK Ltd Wyton, Huntingdon, England
- Age at study initiation: adult
- Weight at study initiation: the animals were weighed immediately before application of the test substance.
- Housing: the rabbits were individually housed in stainless steel cages with flat rod bases or plastica cages with perforated bases, under standardised conventional conditions.
- Diet: Standard diet "Ssniff K4" approx. 100-120g per animal/day; once a day in the morning.
- Water: tap water ad libitum.
- Acclimation period: prior to use the animals were acclimated for approx. one week in the animal room.
- Health check: approx. 24 hours before the test fur was shaved from the dorso-lateral area of the trunk (6x6 cm) of each of the rabbits. Care was taken to avoid abrading the skin. Only animals with healty and intact skin were used.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 1.5 °C
- Humidity: 60-80 %
- Air changes:12-15 times per hour
- Photoperiod: from 6 am to 6 pm, 12 hours artifical illumination.
Vehicle:
unchanged (no vehicle)
Remarks:
pulverized
Controls:
yes, concurrent no treatment
Amount / concentration applied:
100 µl of the pulverised test substance-equivalent to approx. 34 mg
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
21days
Number of animals or in vitro replicates:
3 rabbits
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: treated eye was rinsed with normale saline.
- Time after start of exposure: 24 hours after instillation.

TOOL USED TO ASSESS SCORE: optical instrument (hand slit lamp) and fluorescein. Only effects persisting for more than 24 hours were included in the evaluation.

SCORING SYSTEM
For each animal the Draize scores (cornea, iris, conjunctivae, erythema and swelling and discharge) and the Mcdonald score (aqueous humor) were used. For each animal the Draize scores recorded approx. 24, 48 and 72h after application were added up. The total of these three values was divided by 3 to give the irritation index.
Irritation parameter:
cornea opacity score
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
1
Reversibility:
fully reversible within: 7 days in 2/3 animals
Irritation parameter:
iris score
Basis:
animal: 2/3
Time point:
24/48/72 h
Score:
< 1
Reversibility:
fully reversible within: 7 days in 3/3 animals
Irritation parameter:
conjunctivae score
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
2
Reversibility:
fully reversible within: 21 days in 2/3 animals
Irritation parameter:
chemosis score
Basis:
animal: 3/3
Time point:
24/48/72 h
Score:
< 2
Reversibility:
fully reversible within: 21 days in 3/3 animals
Irritant / corrosive response data:
Exposure of the eye to test substance caused reactions of the mucous membranes and effects of the cornea in all three animals. Additionally, in all three animals the iris was transiently affected and discharge occured. In one animal signs was fully reversible within 21 days, in the second animal corneal signs did not prove to be completely reversible within 21 days and in the third animal a slight erythema was observed on day 21. This evidence indicates that the product may be considered as "severely irritating to the eye".

Individual reactions

Animal Reactions Score at Mean 24/48/72 hrs
1 hr 24 hrs 48 hrs 72 hrs 7 d 14 d 21 d
D11 Corneal opacity 1 1 1 1 0 0 0 1.00
D7 Corneal opacity 1 1 1 1 1* 1+ 1+ 1.00
E8 Corneal opacity 1 1 1* 1* 0 0 0 1.00
D11 Iris 0 0 0 1 0 0 0 0.33
D7 Iris - 1 1 1 0 1 0 1.00
E8 Iris 0 1 0 1 0 0 0 0.67
D11 Conjunctival redness e 2k 2k 2k 0 1 1 2.00
D7 Conjunctival redness e 2 2k 2k 1 1 0 2.00
E8 Conjunctival redness e 2 2k 2k 0 0 0 2.00
D11 Conjunctival chemosis 2v 2 1 1 0 1 0 1.33
D7 Conjunctival chemosis 2v 2 1 1 1 0 0 1.33
E8 Conjunctival chemosis 2v 2 1 1 0 0 0 1.33

e = evaluation not possible due to colouration by test substance

k = conjunctivae and nictitating membrane: injection of blood vessels

v = conjunctivae and nictitating membrane: strong formation of vesicles

* = lower margin of cornea: vascularisation

+ = lower and upper margin of cornea: vascularisation

Interpretation of results:
other: Eye Dam 1 (H318), according to the CLP Regulation (EC) No 1272/2008
Remarks:
Migrated information
Conclusions:
Eye Dam 1, H318
Executive summary:

The irritant/corrosive potential of test substance was studied on the eye of rabbit in accordance with the OECD guideline 405. After gently pulling the lower lid away from the eyeball a volume of 100 µl of the pulverised test substance-equivalent to approx. 34 mg- was placed into the conjunctival sac of one eye of each of three rabbits. 24 hours after instillation of test substance, the treated eye was rinsed with normale saline.

Exposure of the eye to test substance caused reactions on mucous membranes and effects on cornea in all three animals. Additionally in all three animals the iris was transiently affected and discharge occured. In one animal signs was fully reversible within 21 days, in the second animal corneal signs did not prove to be completely reversible within 21 days and in the third animal a slight erythema was observed on day 21. This evidence indicates that the product may be considered as "severely irritating to the eye".

Conclusion

The mean values from gradings at 24, 48 and 72 hours recorded in the study were equal to 1 for corneal opacity in all animals, lower than 1 for iritis in 2 out of three animals, equal to 2 for conjunctival redness and lower than 2 for oedema in all animals. Corneal opacity was not fully recovered within 21 days in one animals, as well as in another animal the conjunctival redness was not recovered withing 21 days.

Due to the fact that at least in one animal effects on the cornea have not fully reversed within an observation period of normally 21 days, a classification as capable to causes serious eye damage (i.e. category 1, H318), according to the CLP Regulation (EC) No 1272/2008, is applied.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

SKIN IRRITATION

The irritant/corrosive potential of Acid Blue 193 was studied on the skin of rabbit, in accordance with OECD guideline 404, under semiocclusive conditions. Evaluation of erythema was not possible in all three animals due to the intense colouration by test substance. Neverthless, no other inflammatory signs (oedema or eschar formation) became apparent within the observation period of 7 days. Signs of toxicity were not seen. This evidence indicates no hazard potential to the skin and the test substance may therefore be regarded as "not irritating" to the skin.

A second experiment conducted following test procedures in accordance with generally accepted scientific standards and well described is available. It was not possible to retreive the detailed composition of tested substance, thus it cannot be confirmed if the composition was comparable to those currently placed on the market; it's only known that it was a concentrated sample. Study outcomes resulted to be consistent with those obtained in the key study.

Very slight erythema was noted at all treated skin sites one hour after treatment and at two treated skin sites at the 24 -hour observation. Very slight oedema was also noted at two treated skin sites at the one-hour observation only. AlI treated skin sites were normal at the 48 and 72-hour observation.

In addition, a study was available on a diluted sample; it has been included for completeness sake. Blue-coloured staining of the skin was noted at all treated skin sites during the observation period. Very slight erythema was noted at one treated skin site one hour after removal of the patches. Very slight or well-defined erythema, with or without very slight oedema, was noted at all treated skin sites at the 24 and 48-hour observations and at two treated skin sites at the 72-hour observation. No evidence of skin irritation was noted at the remaining treated skin site at this time or at any treated skin site on day seven.

EYE IRRITATION

The irritant/corrosive potential of the Acid Blue 193 was studied on the eye of rabbit, in accordance with OECD guideline 405. Exposure of test substance to the eye caused reactions of the mucous membranes and effects of the cornea in all three animals. Additionally, in all three animals the iris was transiently affected and discharge occured. In one animal signs were fully reversible within 21 days, in the second animal corneal signs did not prove to be completely reversible within 21 days and in the third animal a slight erythema was observed on day 21. This evidence indicates that the product may be considered as "severely irritating to the eye".

Also in this case, a second experiment conducted following test procedures in accordance with generally accepted scientific standards and well described is available. It was not possible to retreive the detailed composition of tested substance, thus it cannot be confirmed if the composition was comparable to those currently placed on the market; it's only known that it was a concentrated sample. Study outcomes resulted to be consistent with those obtained in the key study.

Blue staining of the corneal, iridial and conjunctivae tissues was noted in all treated eyes one hour after treatment. Corneal and iridial observations were impeded in one treated eye at the one-hour observation by an extensive blue staining of the tissues. Diffuse, translucent or opalescent corneal opacity was noted in all treated eyes during the observation period. Circumcorneal vascularisation was also apparent in all treated eyes on days 7 and 14 and in two treated eyes on day 21. Iridial inflammation was noted in all treated eyes during the observation period. No iridial inflammation was apparent in any of the treated eyes on day 21. Moderate to severe coniunctivitis was noted in all treated eyes one, 24, 48 and 72 hours after treatment. This condition regressed following the 72-hour observation and minimal coniunctivitis only persisted on days 7 and 14. No evidence of conjunctival irritation was noted on day 21 in any rabbit. A pale appearance of the nictitating membrane was also noted in two of the three treated eyes at the 72-hour observation.

In addition, two studies on a diluted sample of test substance were available; they were included for completeness sake.

In one case, no adverse corneal effects were noted in any treated eye during the observation period. Iridial inflammation was confined to one treated eye at the one-hour observation. Minimal conjunctival irritation was noted in all treated eyes one hour after treatment and in one treated eye at the 24-hour observation. No evidence of conjunctival irritation was noted in the remaining treated eyes at this time or in any treated eye at the 48 and 72-hour observations.

In the second case the complete composition of tested substance was not known, thus a possible contribute to effects by impurity(ies) cannot be excluded. Diffuse corneal opacity, iridial inflammation and moderate conjunctival irritation were noted one hour after treatment; the ocular reactions increased and opalescent or total corneal opacity, iridial inflammation and severe conjunctival irritation was noted at the 72-hour observation. The ocular reactions regressed in two treated eyes that appeared normal on day fourteen. Severe ocular effects including circumcorneal vascularisation persisted in the remaining treated eye on day seven.

Justification for classification or non-classification

According to the CLP Regulation (EC) No 1272/2008, 3.2 Skin corrosion/irritation section, skin irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

Based on the available information, the substance appears as not skin irritating.

According to the CLP Regulation (EC) No 1272/2008, serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance, which is not fully reversible within 21 days of application.

In the key study, the mean values from gradings at 24, 48 and 72 hours recorded in the study were equal to 1 for corneal opacity in all animals, lower than 1 for iritis in 2 out of three animals, equal to 2 for conjunctival redness and lower than 2 for oedema in all animals. Corneal opacity did not fully recover within 21 days in one animals, as well as in another animal the conjunctival redness did not recover within 21 days. Due to the fact that at least in one animal effects on the cornea have not fully reversed within an observation period of normally 21 days, a classification as capable to cause serious eye damage (i.e. category 1, H318), seems to be appropriate.

In conclusion, the substance is not classified for skin irritation; however, a classification as Eye Dam 1 (H318) is proposed, according to the CLP Regulation (EC) No 1272/2008.