Registration Dossier

Administrative data

Description of key information

Skin irritation (similar to OECD TG 404): Irritating
Eye irritation (similar to OECD TG 405): Not irritating

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:
01 May 1984 - 04 May 1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Reliability 2 is assigned because the study is conducted according to an equivalent of OECD TG 404 and in compliance with GLP, but has minor deviations that do not influence the quality of the results.
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
24 instead of 4 hours treatment, no reading at 24 and 72 hours after removal of patch, no 14-day observation period
GLP compliance:
yes
Test material information:
Composition 1
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
- Source: Nottingham University, School of Agriculture, Sutton Bonington, Leics and Rosemead Rabbits Limited, Lea Valley, Essex
- Age at study initiation: 12-16 weeks
- Weight at study initiation: 2.25-3.09 kg
- Housing: individually in suspended metal cages
- Diet: standard laboratory diet (Rabbit Diet, A.W. Tindall Limited), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 60-65
- Air changes (per hr): approximately 10
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
occlusive
Preparation of test site:
other: clipped and abraded (one side)
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated skin of test animals
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
Duration of treatment / exposure:
24 hours
Observation period:
48 hours
Number of animals:
6
Details on study design:
TEST SITE
- Area of exposure: dorsal/flank area, approximately 6 cm2
- Type of wrap if used: Absorbant lint BPC, polythene, adhesive and elasticated corset (Setor's Tubigrip)

REMOVAL OF TEST SUBSTANCE
- Washing: gentle swabbing with cotton wool soaked in lukewarm water
- Time after start of exposure: 24 hours

SCORING SYSTEM:
The irritation was assessed according to the numerical scoring system according to Draize J.H. (1959) Association of Food and Drug Officials of the USA, Austin, Texas, "The Appraisal of the Safety of Chemicals in Food, Drugs and Cosmetics". This is comparable to the current OECD scoring system. Readings were performed at 1 hour and 48 hours after removal of the patch.

A primary cutaneous irritation index was calculated for the test material in the original study. Based on the raw data an erythema and oedema score was calculated for each animal (only intact site), based on the 48 hours reading (after removal of patch). This is the only reading that is available in correspondence with the CLP classification criteria and is therefore used for classification. The 1 hour reading (after removal of patch) provides supportive information.
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: 48 hrs
Score:
3
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
erythema score
Basis:
animal #4
Time point:
other: 48 hrs
Score:
2
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
erythema score
Basis:
animal #5
Time point:
other: 48 hrs
Score:
2
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
erythema score
Basis:
animal #6
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: 48 hrs
Score:
3
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
edema score
Basis:
animal #4
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
edema score
Basis:
animal #5
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritation parameter:
edema score
Basis:
animal #6
Time point:
other: 48 hrs
Score:
4
Max. score:
4
Reversibility:
no data
Remarks on result:
other: score for intact skin
Irritant / corrosive response data:
The test item produced considerable cutaneous irritation in 6 out of 6 rabbits. Well-defined to moderate erythema accompanied by moderate to severe oedema, raised greater than 1 mm and extended well beyond the site of application was observed in all animals after 1 hour (intact and abraded sites). At the 48 hours reading areas of skin necrosis or eschar formation surrounded by a well-defined to moderate erythema and accompanied by severe oedema extending beyond the site of application had developed at the intact and abraded sites of two rabbits and only the intact site of one rabbit. The three remaining rabbits revealed a well-defined to moderate erythema, accompanied by a slight to moderate oedema, although still generally extending beyond the site of application. Additionally skin fissures and desquamation were observed in one rabbit.

Individual scores for skin irritation as indicated in the test report:

Skin reaction Skin site Reading (hours after patch removal) Individual scores
Animal 1 Animal 2 Animal 3 Animal 4 Animal 5 Animal 6
Erythema and eschar formation Intact 1 3 2 2 3 2 3
48 4 4 3 2 2 4
Abraded 1 3 2 2 3 2 3
48 4 4 3 2 2 3
Oedema formation Intact 1 4 4 3 4 4 4
48 4 4 3 2 2 4
Abraded 1 4 4 3 4 4 4
48 4 4 3 2 2 4

Scores for abraded skin are considered not relevant as this type of testing is not included in the current OECD guideline and CLP classification criteria. The 48 hours reading has been used primarily for evaluation of the classification, as this is the only time point available in this study that can be used for classification according to CLP. Skin irritation is indicated if 2/3 of the animals show an erythema or oedema score ≥2.3. The skin irritating effects should be regarded as worst case due to the longer exposure period used in the study (24 instead of 4 hours according to OECD guideline).

At the 48 hour reading, 3 out of 6 animals showed signs related to corrosivity (necrosis on the skin). Taking into account the fact that the exposure of the animals was 24 hours instead of 4 hours, the effects are expected to be present to a larger extent as could be expected from a standard exposure time (OECD). Corrosiveness is indicated according to CLP if 2/3 animals show corrosive reactions (a.o. necrosis). Based on this information, it is anticipated that the observed effects are not sufficient to conclude on corrosive properties of the test substance.

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In a skin irritation study with 6 rabbits, performed similar to OECD 404, erythema scores in 4 out of 6 animals were ≥2.3 and oedema scores in 5 out of 6 animals were ≥2.3 as defined in the EU classification criteria. Based on the results of this study, the substance must be regarded as irritating to skin.
Executive summary:

The undiluted substance (0.5 ml) was applied to 2.5 cm2 skin in a skin irritation test in 6 rabbits with a method similar to OECD TG 404. Deviations consider a 24h instead of a 4h exposure time and only a 1 hour and 48 hour reading (after patch removal), with no prolonged observation time. The score for erythema and oedema from gradings at 48 hours after patch removal was ≥2.3 for a majority of the animals as defined in the EU classification criteria. Moderate to severe irritation was seen, with some signs of necrosis in 3 out of 6 animals. Despite of the necrosis, the substance was not considered corrosive as there was a significantly higher exposure period of 24 instead of the 4 hours (as according to OECD TG 404) used and due to the fact that the effect was not seen in a majority of the animals. Overall the substance must be regarded as irritating to skin.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
2 May 1984 - 5 May 1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Reliability 2 is assigned because the study was conducted according to an acceptable method comparable to OECD TG 405 and in compliance with GLP, but with minor deviations that do not influence the test results.
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
(number of animals used, lack of extended observation period, slight differences in grading system which do not influence the relevance of the results)
Qualifier:
according to
Guideline:
other: Federal Register 1973, Volume 38, No. 187, 1500.42
Deviations:
no
GLP compliance:
yes
Test material information:
Composition 1
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Nottingham University, School of Agriculture, Sutton Bonington, Leics.
- Age at study initiation: 12-16 weeks
- Weight at study initiation: 2.21-2.53 kg
- Housing: individually, in suspended metal cages
- Diet (e.g. ad libitum): standard laboratory diet (Rabbit Diet, A.W. Tindall Limited), ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: minimum five days prior to start of study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3°C
- Humidity (%): 61-65 %
- Air changes (per hr): approximately 10
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: contralateral eye of test animals
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
Single instillation
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM:
Assessment of damage/irritation was made at 24, 48 and 72 hours following treatment according to the numerical evaluation according to Draize J.H. 1959, Assoc. of Food and Drug Officials of the USA, Austin, Texas, "The appraisal of the Safety of Chemical in Foods, Drugs and Cosmetics"). Readings were performed at 24, 48 and 72 48 hours after instillation.

TOOL USED TO ASSESS SCORE:
Examination was done by the use of a standard opthalmoscope (Keeler). Observation of corneal opacity was carried out under ultra-violet illumination (Mineralight UVSL-58) preceded by the instillation into the eye or fluorescein B.P.
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal: #1, #2, #3, #4 and #5
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal #6
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
0.33
Max. score:
4
Irritation parameter:
iris score
Basis:
animal: #1, #2, #3, #4 and #5
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #6
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
0.67
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #4
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
0.67
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal: #1 and #6
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
1.67
Max. score:
3
Reversibility:
not fully reversible within: 72 hours
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal: #2, #3 and #5
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
1
Max. score:
3
Reversibility:
not fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #5
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal: #2, #3 and #4
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
1.33
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #6
Time point:
other: 24, 48 and 72 hrs (mean)
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Irritant / corrosive response data:
The test material produced minimal or slight ocular irritation in 6 out of 6 animals.
- Cornea: diffuse opacity was observed in one rabbit, dulling of the normal lustre of the cornea was seen in two other rabbits (24 hours reading). Dulling was still apparent in one rabbit after 48 hours and 72 hours reading.
- Iris: iritis was observed in one rabbit at 24 and 48 hours reading only.
- Conjunctivae/chemosis: diffuse, crimson-red colouration including slight/distinct swelling of eyelids was observed in 3 rabbits at 24 hours reading. Mild reactions were seen in other 3 animals. At 48 hours, mild inflammation with or without distinct swelling and partial eversion of the eyelid was observed in all animals. This persisted in 4 animals up until the 72 hours reading.
Despite slight irritation at the end of the 72h observation period it is anticipated that the irritation will be resolved within 14 days because of the slight nature of the effect.

Individual scores and individual general totals for ocular irritation:

Rabbit number

1 (619)

2 (413)

3 (419)

4 (427)

5 (429)

6 (431)

Hours after treatment

24

48

72

24

48

72

24

48

72

24

48

72

24

48

72

24

48

72

Cornea

 

E = Degree of opacity

0

0

0

D

0

0

D

0

0

0

0

0

0

0

0

1*

D

D

F = Area of opacity

0

0

0

4

0

0

4

0

0

0

0

0

0

0

0

1

1

1

Score (ExF)x5

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

0

0

Iris

 

D

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1*

1*

0

Score (Dx5)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

5

0

Conjunctivea

 

A = Redness

2*

1

1

2*

1

0

1

1

1

1

1

0

1

1

1

2*

1

1

B = Chemosis

2*

1

1

1

1

0

1

1

0

1

1

0

0

0

0

2*

2*

2*

C = Discharge

1

0

0

1

0

0

1

0

0

1

0

0

0

0

0

3

1

0

Score (A+B+C)x2

10

4

4

8

4

0

6

4

2

6

4

0

2

2

2

14

8

6

Individual general totals

10

4

4

8

4

0

6

4

2

6

4

0

2

2

2

24

13

6

 

GT1

GT2

GT3

GT1

GT2

GT3

GT1

GT2

GT3

GT1

GT2

GT3

GT1

GT2

GT3

GT1

GT2

GT3

* = Positive reaction

D – Dulling of the cornea

Based on the classification systems as included in the study report, the classification of the substance is as follows:

- Classification according to Federal Register 1973:

3/6 rabbits showed positive ocular reactions and the test material therefore gave an INCONCLUSIVE result according to the Federal Register.

 

- E.T.A.D. Classification (Scoring system based on Draize 1959):

The test material produced a Group Mean 3 -Day Score of 5.6 and was therefore classified as NON-IRRITANT to the eye according to ETAD.

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an eye irritation study with rabbits, performed according to a method similar to OECD TG 405, the corneal opacity and/or iritis score were not ≥1 and/or the conjunctival redness and/or oedema (chemosis) score were not ≥2. Based on these results, the substance does not need to be regarded as an eye irritant.
Executive summary:

The potential of Floral Super to cause eye irritation was determined in a test according to a method similar to OECD guideline 405. Six New Zealand White female rabbits were exposed to 0.1 ml of the undiluted test article in one of the eyes. The other eye served as a control. The eyes were examined and the changes graded according to a numerical scale 24, 48 and 72 hours after dosing.

The test material produced minimal or slight ocular irritation in 6 out of 6 rabbits. The corneal opacity and/or iritis score were not ≥1 and/or the conjunctival redness and/or oedema (chemosis) score were not ≥2. Reversibility of the effect within 14 days can be anticipated. Based on these results, the substance does not need to be regarded as an eye irritant.

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

Additional information

In vivo skin irritation

The undiluted substance (0.5 ml) was applied to 2.5 cm2 skin in a skin irritation test in 6 rabbits with a method similar to OECD TG 404. Deviations consider a 24h instead of a 4h exposure time and only a 1 hour and 48 hour reading (after patch removal), with no prolonged observation time. The score for erythema and oedema from gradings at 48 hours after patch removal was ≥2.3 for a majority of the animals as defined in the EU classification criteria. Moderate to severe irritation was seen, with some signs of necrosis in 3 out of 6 animals. Despite of the necrosis, the substance was not considered corrosive as there was a significantly higher exposure period of 24 instead of the 4 hours (as according to OECD TG 404) used and due to the fact that the effect was not seen in a majority of the animals. Overall the substance must be regarded as irritating to skin.

In vivo eye irritation

The potential of Floral Super to cause eye irritation was determined in a test according to a method similar to OECD guideline 405. Six New Zealand White female rabbits were exposed to 0.1 ml of the undiluted test article in one of the eyes. The other eye served as a control. The eyes were examined and the changes graded according to a numerical scale 24, 48 and 72 hours after dosing.

The test material produced minimal or slight ocular irritation in 6 out of 6 rabbits. The corneal opacity and/or iritis score were not ≥1 and/or the conjunctival redness and/or oedema (chemosis) score were not ≥2. Reversibility of the effect within 14 days can be anticipated. Based on these results, the substance does not need to be regarded as an eye irritant.


Justification for selection of skin irritation / corrosion endpoint:
The result of the study is sufficiently reliable and sufficiently adequate for covering the endpoint.

Justification for selection of eye irritation endpoint:
The result of the study is suffienctly reliable and sufficiently adequate for covering the endpoint.

Effects on skin irritation/corrosion: irritating

Justification for classification or non-classification

According to the criteria outlined in Annex VI of 67/548/EEC (DSD) and Annex I of 1272/2008/EC (CLP), the substance does not to be classified as eye irritant, but does need to be classified as skin irritant (Skin Irrit. 2, H315).