Registration Dossier

Administrative data

Description of key information

In vitro skin irritation: Negative
In vivo skin irritation: Negative
In vitro eye irritation (BCOP): No prediction of eye irritation can be made. Cause no serious damage.
In vivo eye irritation: Negative

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:
Testing was conducted between the 10th June 2014 and 13th June 2014
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:
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 3.28 or 3.52 kg and were twelve to twenty weeks old. After an acclimatization period of at least five 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.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., 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 twelve hours continuous light (06:00 to 18:00) and twelve 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.


Justification:
The rabbit is the preferred species of choice as historically used for irritation studies and is specified in the appropriate test guidelines. The number of animals used was the minimum required to achieve the objectives of the study. Testing was conducted in two animals and the response in those animals was such that exposure of a third animal would not affect classification of the test item, no further testing was needed.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Remarks:
The test item was moistened sufficiently with 0.5 ml of distilled water.
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
0.5 g

- Concentration (if solution):
Not applicable.

VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable, for the purpose of the study the test item was used as supplied.

- Concentration (if solution):
Not applicable.

- Lot/batch no. (if required):
Not given in study report.
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
2 females
Details on study design:
TEST SITE
- Area of exposure:
On the day of the test a suitable test site was selected on the back of each rabbit.

- % coverage:
Not given in study report.

- Type of wrap if used:
Test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the shorn skin. 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.

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
Any residual test material removed by gentle swabbing with cotton wool soaked in distilled water.

- Time after start of exposure:
4 hours.

SCORING SYSTEM:
Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the table of evaluation of skin reactions (which can be found in the any other information on methods, including tables section).
Irritation parameter:
erythema score
Basis:
animal: 74371 female
Time point:
other: Mean score at 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No evidence of skin irritation
Irritation parameter:
erythema score
Basis:
animal: 74372 female
Time point:
other: Mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No evidence of skin irritation
Irritation parameter:
edema score
Basis:
animal: 74371 female
Time point:
other: mean of 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No evidence of skin irritation
Irritation parameter:
edema score
Basis:
animal: 74372 female
Time point:
other: mean at 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No evidence of skin irritation
Irritant / corrosive response data:
The individual scores for erythema/eschar and edema are given in Table 1 (please see below in any other information on results section).

Orange colored staining, not preventing evaluation of skin responses, was noted at the test sites of both animals throughout the study.
Other effects:
Both animals showed expected gain in body weight during the study.
Table 1     Individual Skin Reactions:

Skin Reaction

Observation Time
(following patch removal)

Individual Scores –
Rabbit Number and Sex

Total

74371 Female

74372 Female

Erythema/Eschar Formation

Immediately

0STA

0STA

(0 )

1 Hour

0STA

0STA

( 0 )

24 Hours

0STA

0STA

0

48 Hours

0STA

0STA

( 0 )

72 Hours

0STA

0STA

0

Edema Formation

Immediately

0

0

( 0 )

1 Hour

0

0

( 0 )

24 Hours

0

0

0

48 Hours

0

0

( 0 )

72 Hours

0

0

0

Sum of 24 and 72‑Hour Readings (S) :        0

Primary Irritation Index (S/4)              :        0/4 = 0.0

Classification                                       :        NON‑IRRITANT

 


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

STA =   Orange coloured staining

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item produced a primary irritation index of 0.0, when tested on 2 females in an in-vivo skin irritation study (OECD guideline 404). The substance was classified as a non irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.
Executive summary:

Introduction:

The study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit.

 

Results:

A single 4‑Hour, semi‑occluded application of the test item to the intact skin of two rabbits produced no evidence of skin irritation.

 

Conclusion:

The test item produced a primary irritation index of 0.0 and was classified as non‑irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

 

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.

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:
Study conducted between 9th September 2014 and 25th September 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
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:
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.96 or 3.23 kg and were twelve to twenty weeks old. After an acclimatization period of at least five 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.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., 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 twelve hours continuous light (06:00 to 18:00) and twelve 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.

Justification
The rabbit is the preferred species of choice as historically used for irritation studies and is specified in the appropriate test guidelines. The number of animals used was the minimum required to achieve the objectives of the study. Testing was conducted in two animals and the response in those animals was such that exposure of a third animal would not affect classification of the test item, therefore, no further testing was needed.

Vehicle:
unchanged (no vehicle)
Remarks:
For the purpose of the study the test item was used as supplied.
Controls:
other: EXAMPLE: The left eye remained untreated and was used for control purposes.
Amount / concentration applied:
TEST MATERIAL

- Amount(s) applied (volume or weight with unit):
A volume of 0.1 mL of the test item, which was found to weigh approximately 96 mg (as measured by gently compacting the required volume into an adapted syringe) 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

VEHICLE
Not applicable, for the purpose of the study the test item was used as supplied.
Duration of treatment / exposure:
72 hours
Observation period (in vivo):
Approximately 1 hour, 24, 48 and 72 hours and 7 days following treatment.
Number of animals or in vitro replicates:
2 males
Details on study design:
PRE STUDY:
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.

TESTING PROCEDURE:
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.

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.

Observations were made, 1, 24, 48 and 72 hours. Any clinical signs of toxicity, if present, were also recorded and bodyweights were recorded too. An additional observation was made in one treated eye on Day 7 to assess the reversibility of the ocular effects.

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
Not applicable
- Time after start of exposure:
Not applicable

SCORING SYSTEM:
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the draize scale which can be found from (Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49). The Draize scoring system has been included in the any other information on methods section (including tables).

TOOL USED TO ASSESS SCORE:
Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Irritation parameter:
cornea opacity score
Basis:
animal: 74708 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
cornea opacity score
Basis:
animal: 74726 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
iris score
Basis:
animal: 74708 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: no effects observed
Irritation parameter:
iris score
Basis:
animal: 74726 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: no effects observed
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 74708 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
1.66
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
other: redness
Basis:
animal: 74726 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
0.66
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal: 74708 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal: 74726 Male
Time point:
other: mean 24, 48 and 72 hours
Score:
0.661
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritant / corrosive response data:
Individual and group mean scores for ocular irritation are given in Table 1*

Orange colored staining of the fur around the treated eye was noted in both animals at all observations.

No corneal or iridial effects were noted during the study.

Moderate conjunctival irritation was noted in both treated eyes 1 hour after treatment. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in the other treated eye at the 24 and 48 Hour observations. Minimal conjunctival irritation was noted in one treated eye at the 72 Hour observation.

One treated eye appeared normal at the 72 Hour observation and the other treated eye appeared normal at the 7 Day observation.

* Table 1 can be found in the any other information on results (including tables section of the robust study summary).
Other effects:
Body weights:
Both animals showed expected gain in body weight during the study.

Table 1     Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

74708Male

74726Male

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

CORNEA

 

 

 

 

 

 

 

 

 

E = Degree of Opacity

0

0

0

0

0

0

0

0

0

F = Area of Cornea Involved

0

0

0

0

0

0

0

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

0

IRIS

 

 

 

 

 

 

 

 

 

D

0

0

0

0

0

0

0

0

0

Score (D x 5)

0

0

0

0

0

0

0

0

0

CONJUNCTIVAE

 

 

 

 

 

 

 

 

 

A = Redness

2

2

2

1

0

2

1

1

0

B = Chemosis

2

2

1

1

0

1

1

1

0

C = Discharge

2Sf

2Sf

1Sf

1Sf

0Sf

1Sf

1Sf

0Sf

0Sf

Score (A + B + C) x 2

12

12

8

6

0

8

6

4

0

Total Score

12

12

8

6

0

8

6

4

0

IPR=Initial pain reaction

Sf =        Orange colored staining of the fur around the treated eye

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.
Executive summary:

Introduction:

The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit.

 

Results:

A single application of the test item to the non-irrigated eye of two rabbits produced moderate conjunctival irritation. One treated eye appeared normal at the 72‑Hour observation and the other treated eye appeared normal at the 7‑Day observation.

 

Conclusion: 

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.

 

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Firstly an in vitro skin irritation test was performed according to OECD 439 and the conclusion was that the test item was classified as non-irritant. The following classification criteria apply:

EU DSD & CLP Not classified for Irritation.
UN GHS Not classified for Irritation (category 3 can not be determined).

In the in vivo skin irritation performed on rabbits according to the OECD 404 (Dermal irritation/corrosion, the test item produced a primary irritation index of 0.0 and was classified as non‑irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

In the BCOP experiment (OECD 437) performed with FAT 40863/A TE, the conclusion was:

No prediction of eye irritation can be made.

The results of this study have identified the test item as not causing serious eye damage, but they do not permit conclusion that the test item does not require classification for eye irritation.

Then an in vivo experiment (OECD 405) was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit.

A single application of the test item to the non-irrigated eye of two rabbits produced moderate conjunctival irritation. One treated eye appeared normal at the 72‑Hour observation and the other treated eye appeared normal at the 7‑Day observation.

 FAT 40863/A TE, does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.


Justification for selection of skin irritation / corrosion endpoint:
A skin in vitro irritation test was performed and was negative. The in vivo test is also negative, then as key study we select the in vivo test.

Justification for selection of eye irritation endpoint:
The results of the BCOP test have identified FAT 40863/A TE as not causing serious eye damage, but they do not permit conclusion that the test item does not require classification for eye irritation.
The in vivo test performed on rabbit concluded that FAT 40863/A TE is not irritating to rabbit eyes.

Justification for classification or non-classification

Based on the above mentioned results the substance does not need to be classified according to CLP regulation (Regulation EC No.1272/2008) and DSD (Directive 67/548/EEC).