Registration Dossier

Administrative data

Description of key information

The skin irritation/corrosion potential of the test substance, IFF 215 (Floriane) was assessed according to the following guideline studies:

OECD Test Guideline 439 using an in vitro skin irritation: Reconstructed Human Epidermis Model Test: The test item was considered to be not classified as an irritant.

OECD Test Guideline 431 using a Reconstructed Human EpiDermis (RHE): The test item was considered to be non-corrosive to the skin.

OECD Test Guideline 404 using the Acute Dermal Irritation/Corrosion Method: The test item was classified as a Category 2 (irritant) based on GHS criteria. No corrosive effects were noted.

The eye irritation potential of the test substance, IFF 215 (Floriane) was assessed according to the following guideline studies:

OECD Test Guideline 437 using the Bovine Corneal Opacity and Permeability (BCOP) Assay (2016): No prediction of eye irritation could be made.

OECD Test Guideline 492 using the In vitro Eye Irritation Test: Reconstructed Human EpiocularTM Model: The test item was considered to be non-irritant to the eye.

OECD Test Guideline 405 Acute Eye Irritation/Corrosion method: The test item was considered to be non-irritant to the eye.

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:
The study was conducted between 17 March 2016 and 31 March 2016.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study is considered to be a reliability 1 as it has been conducted according to OECD Test Guideline 404 using the Acute Dermal Irritation/Corrosion Method and in compliance with GLP.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Animal Information
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Envigo RMS (UK) Limited, Leicestershire, UK. At the start of the study the animals weighed 2.86 or 3.07 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.

Animal Care and Husbandry
The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, 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 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.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
On the day of the test a suitable test site was selected on the back of each rabbit. A quantity of 0.5 mL of the test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in position with a strip of surgical adhesive tape. To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.
Duration of treatment / exposure:
Four hours.
Observation period:
1, 24, 48 and 72 hours.
Number of animals:
Two male rabbits.
Details on study design:
Study Design
On the day before the test each of a group of two rabbits was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.
On the day of the test a suitable test site was selected on the back of each rabbit. A quantity of 0.5 mL of the test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in position with a strip of surgical adhesive tape. To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.
Four hours after application the corset and patches were removed from each animal and any residual test item removed by gentle swabbing with cotton wool soaked in 74% Industrial Methylated Spirit.
Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the following scale:

EVALUATION OF SKIN REACTIONS
Erythema and Eschar Formation Value
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema 4

Edema Formation
No edema 0
Very slight edema (barely perceptible) 1
Slight edema (edges of area well-defined by definite raising) 2
Moderate edema (raised approximately 1 millimeter) 3
Severe edema (raised more than 1 millimeter and extending beyond the area of exposure) 4

Any other skin reactions and clinical signs of toxicity, if present, were also recorded.
Additional observations were made on Days 7 and 14 to assess the reversibility of skin reactions.
Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Data Evaluation
Calculation of Primary Irritation Index and Grading of Irritancy Potential Using the Draize Scheme
The scores for erythema and edema at the 24 and 72 Hour readings were totaled for the two test rabbits (8 values) and this total was divided by 4 to give the primary irritation index of the test item. The test item was classified according to the following scheme devised by Draize, J.H. (1959):

Primary Irritation Index Classification of Irritancy
0 Non-irritant
> 0 to 2 Mild irritant
> 2 to 5 Moderate irritant
> 5 to 8 Severe irritant

If irreversible alteration of the dermal tissue is noted in any rabbit, as judged by the Study Director, which include ulceration and clear necrosis or signs of scar tissue, the test item is classified as corrosive to rabbit skin. Classification according to Draize may, therefore, not be applicable.

Major Computerized Systems
The following computerized system was used in the study:
Delta Controls – ORCAview (Version 3.4.0)
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
75393 Male
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
probability of moderate irritation
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
75383 Male
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
probability of moderate irritation
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
75393 Male
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
probability of moderate irritation
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
75383 Male
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
probability of moderate irritation

 Skin Reactions

The individual scores for erythema/eschar and edema are given in the following table:

Individual Skin Reactions

Skin Reaction

Observation Time
(following patch removal)

Individual Scores

Total

Rabbit Number and Sex

75393Male

75383Male

Erythema/Eschar Formation

Immediately

1

1

(2 )

1 Hour

1

1

( 2 )

24 Hours

2

2

4

48 Hours

2

2

( 4 )

72 Hours

2

2

4

7 Days

?eD+

0D

( 0-4 )

14 Days

0

0

( 0 )

Edema Formation

Immediately

0

0

( 0 )

1 Hour

1

1

( 2 )

24 Hours

2

2

4

48 Hours

2

2

( 4 )

72 Hours

2

2

4

7 Days

?od

0

( 0-4 )

14 Days

0

0

( 0 )

Sum of 24 and 72‑Hour Readings (S)            :              16

Primary Irritation Index (S/4)                          :              16/4 = 4.0

Classification                                                      :              MODERATEIRRITANT

 


(   ) =    Total values not used for calculation of primary irritation index

D+ =      Severe desquamation

D =         Moderate desquamation

?e =        Presence of adverse dermal reactions (not including staining) precludes macroscopic observation of erythema

?od =     Degree of thickening/scabbing is such that a macroscopic observation of edema is not possible

Very slight erythema was noted at both treated skin sites immediately after patch removal with very slight erythema and very slight edema noted 1 hour after patch removal. Well‑defined erythema and slight edema were noted at both treated skin sites at the 24, 48 and 72‑Hour observations. 

Severe desquamation, preventing accurate evaluation of erythema and edema, was noted at one treated skin site with moderate desquamation noted at the other treated skin site at the 7‑Day observation.

Both treated skin sites appeared normal at the 14‑Day observation.

Body Weight

Individual body weights and body weight change are given in the following table:

Individual Body Weights and Body Weight Change

Rabbit Number
and Sex

Individual Body Weight (kg)

Body Weight Change (kg)

Day 0

Day 14

75393Male

3.07

3.45

0.38

75383Male

2.86

3.11

0.25

 

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

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
The test item produced a primary irritation index of 4.0 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.
Executive summary:

The skin irritation potential of the test substance, IFF 215 (Floriane) was assessed according to OECD Test Guideline 404 using the acute dermal irritation/corrosion Method. The test material was classified as a moderate irritant to rabbit skin according to the Draize classification scheme and classified as a Category 2 (irritant) based on GHS criteria. No corrosive effects were noted.

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:
The study was conducted between 23 May 2016 and 07 June 2016.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study is considered to be a reliability 1 as it has been conducted according to OECD Test Guideline 405 Acute Eye Irritation/Corrosion method and in compliance with GLP.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute 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:
Animal Information
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Envigo RMS (UK) Limited, Leicestershire, UK. At the start of the study the animals weighed 2.94 or 4.40 kg and were 12 to 52 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.

Animal Care and Husbandry
The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, 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.

ENVIRONMENTAL CONDITIONS
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 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:
yes, concurrent negative control
Amount / concentration applied:
A volume of 0.1 mL of the test item was placed into the conjunctival sac of the right eye.

Duration of treatment / exposure:
72 hours
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
2 male rabbits
Details on study design:
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 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 - Initial Pain Reaction, please see "Any other information on materials and methods, inc. tables" for further information.
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 below.
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.
An additional observation was made on Day 7 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.

Draize Scale for Scoring Ocular Irritation
1. CONJUNCTIVAE Score
(A) Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3
(B) Chemosis
No swelling 0
Any swelling above normal (includes nictitating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids half closed to completely closed 4
(C) Discharge
No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs a considerable area around the eye 3
THE TOTAL SCORE = (A + B + C) x 2 MAXIMUM TOTAL = 20

2. IRIS
(D) Values
Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof)
iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, haemorrhage, gross destruction (any or all of these) 2
THE TOTAL SCORE = D x 5 MAXIMUM TOTAL = 10

3. CORNEA
(E) Degree of Opacity (most dense area used)
No opacity 0
Scattered or diffuse areas, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris not discernible through the opacity 4
(F) Area of Cornea Involved
One quarter (or less) but not zero 1
Greater than one quarter but less than half 2
Greater than half but less than three quarters 3
Greater than three quarters, up to whole area 4
THE TOTAL SCORE = (E x F) x 5 MAXIMUM TOTAL = 80
MAXIMUM TOTAL SCORE POSSIBLE = 110
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.66
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
Ocular Reactions
Please see "Any other information on results, incl. tables" for tabulated data.
No corneal effects were noted during the study.
Iridial inflammation was noted in one treated eye 1 hour after treatment and in the other treated eye at the 24-Hour observation.
Moderate conjunctival irritation was noted in both treated eyes 1 and 24 hours after treatment with minimal conjunctival irritation noted at the 48 and 72-Hour observations.
Both treated eyes appeared normal at the 7-Day observation.

Ocular Reactions

Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

5464Male

75443 Male

IPR = 0

IPR = 0

Time After Treatment

1 Hour

24 Hours

48 Hours

72 Hours

7 Days

1 Hour

24 Hours

48 Hours

72 Hours

7 Days

Cornea

0

0

0

0

0

0

0

0

0

0

E = Degree of Opacity

0

0

0

0

0

0

0

0

0

0

F = Area of Cornea Involved

0

0

0

0

0

0

0

0

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

0

0

Iris

 

 

 

 

 

 

 

 

 

 

D

1

0

0

0

0

0

1

0

0

0

Score (D x 5)

5

0

0

0

0

0

5

0

0

0

Conjunctivae

 

 

 

 

 

 

 

 

 

 

A = Redness

2

2

1

1

0

2

2

1

1

0

B = Chemosis

2

1

1

0

0

2

1

1

1

0

C = Discharge

2

1

1

0

0

2

1

0

0

0

Score (A + B + C) x 2

12

8

6

2

0

12

8

4

4

0

Total Score

17

8

6

2

0

12

13

4

4

0

IPR = Initial pain reaction

Individual Total Scores and Group Mean Scores for Ocular Irritation

Rabbit Number and Sex

Individual Total Scores At:

1 Hour

24 Hours

48 Hours

72 Hours

7 Days

75464 Male

17

8

6

2

0

75443 Male

12

13

4

4

0

Group Total

29

21

10

6

0

Group Mean Score

14.5

10.5

5.0

3.0

0.0

Body Weight

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

Interpretation of results:
GHS criteria not met
Conclusions:
The test item produced a maximum group mean score of 14.5 and was classified as a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.
The test item does not meet the criteria for classification according to the Globally Harmonized System of Classification and Labelling of Chemicals or to Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.
Executive summary:

The eye irritation potential of the test item, IFF 215 (Floriane), was assessed according to OECD Test Guideline 405 using the Acute Eye Irritation/Corrosion method. According to the conditions of the test, the test item was considered to be non-irritant to the eye.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Acute Dermal Irritation

Substances can be allocated to one of two skin irritation/corrosion categories based on the skin irritation/corrosion route according to the Globally Harmonized Classification System and Regulation (EC) No. 1272/2008, relating to the classification, Labelling and Packaging of Substances and Mixtures.

Skin irritation/corrosion is assessed based on effects noted during testing with classification being determined by the severity of the effects.

A test substance is considered to be corrosive if up to 4 hours after application it produces irreversible damage to the skin. Classification is further sub-divided based on the severity of the corrosive effects.

A test substance is considered to be irritant if up to 4 hours after application it produces reversible damage to the skin.

Three studies were carried out, two in vitro and one in vivo, to determine the skin irritation/corrosion potential of the test material IFF 215 (Floriane).

In assessing the potential for skin irritation/corrosion, in vitro testing is recommended initially. As the in vitro studies indicated that the substance was not considered an irritant or corrosive, additional in vivo testing was then carried out. The subsequent in vivo testing provided results indicating that the test material should be considered to be an irritant. Although in vitro data is considered to be acceptable for classification purposes, in vivo data is considered to be more reliable for classification and as such as in vivo results were available, these were used for the final classification decision.

Based on the results of the in vivo study carried out according to OECD 404, it can be concluded that the test material IFF 215 (Floriane) is classified as a Category 2 (irritant) based on GHS criteria.

Eye Irritation

Substances can be allocated to one of two serious eye damage/irritation categories based on the serious eye damage/irritation route according to the Globally Harmonized Classification System and Regulation (EC) No. 1272/2008, relating to the classification, Labelling and Packaging of Substances and Mixtures.

Serious eye damage/irritation is assessed based on effects noted during testing with classification being determined by the severity of the effects.

A test substance is considered to cause serious eye damage after application to the anterior surface of the eye if it produces irreversible tissue damage and/or serious physical decay in vision.

A test substance is considered to be an irritant if after application to the anterior surface of the eye it produces changes that are fully reversible within 21 days.

Three studies were carried out, two in vitro and one in vivo, to determine the eye irritation potential of the test material IFF 215 (Floriane).

In assessing the potential for eye irritation, in vitro testing is recommended initially. As one of the in vitro studies could not predict the eye irritation potential of the test substance and the second in vitro study indicated that the substance was not considered an irritant, additional in vivo testing was then carried out. The subsequent in vivo testing provided results indicating that the test material should not be considered to be an irritant. Although in vitro data is considered to be acceptable for classification purposes, in vivo data is considered to be more reliable for classification and as such as in vivo results were available, these were used for the final classification decision.

Based on the results of the in vivo study carried out according to OECD 405, it can be concluded that the test material IFF 215 (Floriane) is not classified as an irritant to the eye based on GHS criteria.