Registration Dossier

Diss Factsheets

Administrative data

Description of key information

In vitro skin corrosion (OECD 431): Negative
In vitro skin irritation (OECD 439): Negative
Acute dermal irritation in rabbit (OECD 404): Negative
In vitro eye irritation (BCOP, OECD 437): Inconclusive due to the staining of the test item treated corneas precluding accurate evaluation.
In vivo eye irritation (OECD 405): 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 8th and 11th July 2014.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system 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 2.59 or 2.83 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:
other: clipped
Vehicle:
other: Test material was moistened with 0.5 ml of distilled water.
Controls:
no
Amount / concentration applied:
TEST MATERIAL
On the day of the test a suitable test site was selected on the back of each rabbit. At each test site a quantity of 0.5 g of the test item, moistened sufficiently with 0.5 mL of distilled water to achieve a paste, was introduced 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.

VEHICLE
Not applicable
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
2
Details on study design:
Procedure:
On the day before the test two rabbits were 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. At each test site a quantity of 0.5 g of the test item, moistened sufficiently with 0.5 mL of distilled water to achieve a paste, was introduced 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 distilled water.

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.

Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.


Irritation parameter:
erythema score
Remarks:
Erythema/Eschar Formation
Basis:
animal: 74478 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
Remarks:
Erythema/Eschar Formation
Basis:
animal: 74479 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:
other: Oedema Formation
Basis:
animal: 74478 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:
other: Oedema Formation
Basis:
animal: 74479 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:
No evidence of skin irritation was noted during the study. Please see table 1 in the any other information on results section for more details.
Other effects:
Violet colored staining, not preventing evaluation of skin responses, was noted at both treated skin sites during the study.

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

Table 1:

Skin Reaction

Observation Time
(following patch removal)

Individual Scores – Rabbit Number and Sex

Total

74478Female

74479Female

Erythema/Eschar Formation

Immediately

0STA

0STA

(0 )

1 Hour

0STA

0STA

( 0 )

24 Hours

0

0STA

0

48 Hours

0

0STA

( 0 )

72 Hours

0

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 =    Violet colored 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 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.
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 of0.0and 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 was conducted between the 09 June 2014 and 10th July 2014.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Three 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 2.46 to 3.27 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.
Vehicle:
unchanged (no vehicle)
Controls:
other: 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 material was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball.

- Concentration (if solution):
Undiluted and used as supplied

VEHICLE
Not applicable - test material was used as supplied
Duration of treatment / exposure:
72 hours
Observation period (in vivo):
Approximately 1 hour, 24, 48, 72 hours and 7 days following treatment.
Number of animals or in vitro replicates:
3 animals were tested in total. (After consideration of the ocular responses produced in the first treated animal, two additional animals were treated).
Details on study design:
PROCEDURE:
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, which was found to weigh approximately 86 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. Immediately after administration of the test item, an assessment of the initial pain reaction was made.

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, two additional animals were similarly treated.

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. For more details the draize scale can be found in the any other information on results section.

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: 74365 Female
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: 74431 female
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks:
No effects observed
Irritation parameter:
cornea opacity score
Basis:
animal: 74467 female
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: 74365 female
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
iris score
Basis:
animal: 74431 female
Time point:
other: Mean 24, 48 and 72 hours
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal: 74467 female
Time point:
other: Mean 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
other: redness
Basis:
animal: 74365 Male
Time point:
other: Mean 24, 48 and 72 hours
Score:
0.66
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
other: redness
Basis:
animal: 74431 female
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: 74467 female
Time point:
other: Mean 24, 48 and 72 hours
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal: 74365 female
Time point:
other: Mean 24, 48 and 72
Score:
0.66
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal: 74431 female
Time point:
other: Mean 24, 48 and 72 hours
Score:
1.66
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal: 74467 female
Time point:
other: 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritant / corrosive response data:
Ocular Reactions:
Individual and group mean scores for ocular irritation are given in Table 1 and Table 2*.

Black staining of the cornea, iris and conjunctival membranes was noted in all treated eyes 1 hour after treatment and prevented accurate evaluation of corneal, iridial and conjunctival redness effects.

Black staining of the fur was noted around the treated eye of two animals at all observations.

No corneal effects were noted at the 24-Hour observation and at subsequent observations.

Iridial inflammation was noted in one treated eye at the 24, 48 and 72 Hour observations.

Moderate conjunctival irritation was noted in all treated eyes at the 1 and 24 Hour observations. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in two treated eyes at the 48 Hour observation. Minimal conjunctival irritation was noted in one treated eye at the 72 Hour observation.

Two treated eyes appeared normal at the 72 Hour observation and one treated eye appeared normal at the 7 Day observation.

* All tables can be found in the any other information on results section.
Other effects:
All animals showed expected gain in body weight during the study.

TABLES:

Table 1     Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex

74365Female

74431Female

74467Female

IPR= 0

IPR = 0

IPR = 0

Time After Treatment

1
Hour

24
Hours

48
Hours

72
Hours

1
Hour

24
Hours

48
Hours

72
Hours

7
Days

1
Hour

24
Hours

48
Hours

72
Hours

CORNEA

 

 

 

 

 

 

 

 

 

 

 

 

 

E = Degree of Opacity

?s

0

0

0

?s

0

0

0

0

?s

0

0

0

F = Area of Cornea Involved

?s

0

0

0

?s

0

0

0

0

?s

0

0

0

Score (E x F) x 5

0-80

0

0

0

0-80

0

0

0

0

0-80

0

0

0

IRIS

 

 

 

 

 

 

 

 

 

 

 

 

 

D

?s

0

0

0

?s

1

1

1

0

?s

0

0

0

Score (D x 5)

0-10

0

0

0

0-10

5

5

5

0

0-10

0

0

0

CONJUNCTIVAE

 

 

 

 

 

 

 

 

 

 

 

 

 

A = Redness

?s

1

1

0

?s

2

2

1

0

?s

2

1

0

B = Chemosis

2

1

1

0

2

2

2

1

0

2

2

1

0

C = Discharge

2Sf

2Sf

1Sf

0Sf

2Sf

1Sf

1Sf

1Sf

0Sf

2

1

1

0

Score (A + B + C) x 2

8-14

8

6

0

8-14

10

10

6

0

8-14

10

6

0

Total Score

8-104

8

6

0

8-104

15

15

11

0

8-104

10

6

0

IPR=Initial pain reaction

Sf =        Black staining of fur around treated eye

?s =        Degree of ocular staining prevented accurate evaluation of reactions

- =         Observation not required - considered to be zero for calculation of Group Mean Score

 

Table 2     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

74365Female

8-104

8

6

0

-

74431Female

8-104

15

15

11

0

74467Female

8-104

10

6

0

-

Group Total

24-312

33

27

11

0

Group Mean Score

8-104

11.0

9.0

3.7

0.0

 



Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item produced a group mean score in the range of 8.0 to 104.0 and was considered to be at least 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 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 three rabbits produced iridial inflammation and moderate conjunctival irritation. Thedegree of ocular staining prevented accurate the evaluation of the cornea, iris and conjunctival redness 1 hour after treatment. Two treated eyes appeared normal at the 72‑Hour observation and one treated eye appeared normal at the 7‑Day observation.

 

Conclusion:

The test item produced a group mean score in the range of 8.0 to 104.0 and was considered to be at least 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 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 , two in vitro test on the skin were performed (OECD 431 and OECD 439).

FAT 20341/A TE was classified as non-corrosive to the skin (OECD 431). The following classification criteria apply:

EU DSD (67/548/EEC): Not classified for corrosivity.

EU CLP (1272/2008/EC)/UN GHS: Not classified for corrosivity.

UN Packing Group: Non-Corrosive.

FAT 20341/A TE was classified as non-irritant to the skin for the OECD 439. 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 20341/A TE, the conclusion was:

No prediction of eye irritation can be made. The results of this test are considered to be inconclusive due to the staining of the test item treated corneas precluding accurate evaluation.

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 20341/A TE produced a group mean score in the range of 8.0 to 104.0 and was considered to be at least a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. FAT 20341/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:
Results from both in vitro studies (in vitro skin corrosion (OECD 431) and in vitro skin irritation (OECD 439)) are negative.
The in vivo test is also negative (OECD 404), then as key study we select the in vivo test.

Justification for selection of eye irritation endpoint:
Results from the in vitro study (BCOP, OECD 437) are considered to be inconclusive due to the staining of the test item treated corneas precluding accurate evaluation.
The in vivo test is also negative (OECD 405), then as key study we select the in vivo test.

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).