Registration Dossier

Administrative data

Description of key information

Acute irritation studies in the rabbit show moderate skin and slight eye irritation (OECD 404 and 405, respectively). However, teh substance is not classified.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Remarks:
The study has been conducted according to guideline OECD TG 404.
Type of information:
experimental study
Adequacy of study:
key study
Study period:
between 23 July 2008 and 26 August 2008.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
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)
Remarks:
Date of inspection: 21/08/2007 Date of signature: 15/10/07
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
Harlan UK Limited, Bicester, Oxon, UK

- Age at study initiation:
twelve to twenty weeks old

- Weight at study initiation:
At the start of the study the animals were in the weight range of 2.0 to 3.5 kg

- Housing:
The animals were individually housed in suspended cages

- Diet (e.g. ad libitum):
Free access to food (2030 Teklad Global Rabbit diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK) was allowed throughout the study

- Water (e.g. ad libitum):
Free access to mains drinking water was allowed throughout the study

- Acclimation period:
An acclimatisation period of at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
The temperature was set to achieve limits of 17 to 23°C

- Humidity (%):
The relative humidity was set to achieve limits of 30 to 70%

- Air changes (per hr):
The rate of air exchange was at least fifteen changes per hour

- Photoperiod (hrs dark / hrs light):
lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.


IN-LIFE DATES: From: Day 1 To: Day 14
Type of coverage:
semiocclusive
Preparation of test site:
other: clipped free of fur from the dorsal/flank area using veterinary clippers.
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
At each test site a quantity of 0.5 ml of the test material was introduced

- Concentration (if solution):
Not applicable

VEHICLE
Not applicable
Duration of treatment / exposure:
One patch was applied to the back of each rabbit, and was allowed to remain in contact with the skin for a period of four hours.
Observation period:
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

Additional observations were made on Days 7 and 14 to assess the reversibility of skin reactions.
Number of animals:
One rabbit was initially treated

After consideration of the skin reactions produced in the first animal, two additional animals were treated
Details on study design:
TEST SITE
- Area of exposure:
On the back of the rabbits

- % coverage:
Not recorded

- Type of wrap if used:
The test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the shorn skin. Each patch was secured in position with a strip of surgical adhesive tape. To prevent the animal interfering with the patches, the trunk of the rabbit was wrapped in an elasticated corset for the duration of the exposure period


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

- Time after start of exposure:
Four hours


SCORING SYSTEM:
See Table A

Interpretation of Results
Calculation of Primary Irritation Index and Grading of Irritancy Potential Using the Draize Scheme

The scores for erythema and oedema at the 24 and 72-hour readings were totalled for the three test rabbits (12 values) and this total was divided by six to give the primary irritation index of the test material. The test material was classified according to the following scheme devised by Draize J H (1959) "Dermal Toxicity" In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Association of Food and Drug Officials of the United States, Austin, Texas, p.46 59:

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 material is classified as corrosive to rabbit skin. Classification according to Draize may, therefore, not be applicable.
Irritation parameter:
erythema score
Remarks:
Erythema/Eschar Formation
Basis:
animal: 67624 Male
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 14 days.
Remarks on result:
other: Light brown discolouration of the epidermis, Loss of skin elasticity and Loss of skin flexibility at 24, 48 and 72 hours. Crust formation at 7 days.
Irritation parameter:
edema score
Remarks:
Oedema Formation
Basis:
animal: 67624 male
Time point:
24/48/72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 7 days.
Irritation parameter:
erythema score
Remarks:
Erythema/Eschar Formation
Basis:
animal: 67679 male
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days.
Remarks on result:
other: Light brown discolouration of the epidermis, Loss of skin elasticity and Loss of skin flexibility at 48 and 72 hours. Crust formation at 7 days.
Irritation parameter:
edema score
Remarks:
Oedema Formation
Basis:
animal: 67679 male
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
Erythema/Eschar Formation
Basis:
animal: 67680 male
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: Light brown discolouration of the epidermis, Loss of skin elasticity and Loss of skin flexibility at 48 and 72 hours. Crust formation at 7 days.
Irritation parameter:
edema score
Remarks:
Oedema Formation
Basis:
animal: 67680 male
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
Skin Reactions
3-Minute Exposure Period
The individual scores for erythema/eschar and oedema are given in Table 1.

Well-defined erythema and very slight oedema were noted at the treated skin site at the 24, 48 and 72-hour observations. Loss of skin elasticity was also noted at the treated skin site at the 72 hour observation and moderate desquamation was noted at the treated skin site at the 7-day observation.

The treated skin site appeared normal at the 14-day observation.

1-Hour Exposure Period
The individual scores for erythema/eschar and oedema are given in Table 1.

Well-defined erythema and very slight oedema were noted at the treated skin site at the 24, 48 and 72-hour observations. Loss of skin elasticity was also noted at the treated skin site at the 72 hour observation and crust formation was noted at the treated skin site at the 7-day observation.

The treated skin site appeared normal at the 14-day observation.

4-Hour Exposure Period
The individual scores for erythema/eschar and oedema are given in Table 2.

Well-defined erythema and very slight oedema were noted at all treated skin sites one hour after patch removal and at the 24, 48 and 72-hour observations with very slight erythema noted at the 7 day observation. Light brown discolouration of the epidermis and loss of skin flexibility and/or elasticity were noted at one treated skin site at the 24 hour observation and at all treated skin sites at the 48-hour and 72-hour observations. Crust formation was noted at all treated skin sites at the 7 day observation.

All treated skin sites appeared normal at the 14-day observation.
Other effects:
Bodyweight
Individual bodyweights and bodyweight changes are given in Table 3.
All animals showed expected gain in bodyweight during the study.

Table1               Individual Skin Reactions Following 3-Minute and 1-Hour Exposures

Skin Reaction

Observation Time

Individual Scores - Rabbit Number and Sex

67624Male

3-Minute Exposure

1-Hour Exposure

Erythema/Eschar Formation

1 Hour

0

0

24 Hours

2

2

48 Hours

2

2

72 Hours

2Le

2Le

7 Days

0D

0Cf

14 Days

0

0

Oedema Formation

1 Hour

0

0

24 Hours

1

1

48 Hours

1

1

72 Hours

1

1

7 Days

0

0

14 Days

0

0


D= Moderate desquamation

Le = Loss of skin elasticity

Cf = Crust formation

Table2               IndividualSkin ReactionsFollowing 4-Hour Exposure

Skin Reaction

Observation Time

Individual Scores – Rabbit Number and Sex

Total

67624Male

67679Male

67680Male

Erythema/Eschar Formation

1 Hour

2

2

2

(6 )

24 Hours

2BrLeLf

2

2

6

48 Hours

2BrLeLf

2BrLeLf

2BrLeLf

( 6 )

72 Hours

2BrLeLf

2BrLeLf

2BrLeLf

6

7 Days

1Cf

1Cf

1Cf

( 3 )

14 Days

0

0

0

( 0 )

Oedema Formation

1 Hour

1

1

1

( 3 )

24 Hours

1

1

1

3

48 Hours

1

1

1

( 3 )

72 Hours

1

1

1

3

7 Days

0

0

0

( 0 )

14 Days

0

0

0

( 0 )

Sum of 24 and 72-hour Readings (S):              18

Primary Irritation Index (S/6)                             :              18/6 = 3.0

Classification                                                       :              MODERATE IRRITANT


Introduction. The study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

§        OECD Guidelines for the Testing of Chemicals No. 404 “Acute Dermal Irritation/Corrosion” (adopted)

§        Method B4 Acute Toxicity (Skin Irritation) of Commission Directive 2004/73/EC

Results. 3-minute and 1-hour semi-occluded applications of the test material to the intact skin of one rabbit produced no corrosive effects.

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced well-defined erythema and very slight oedema. Other skin reactions noted were light brown discolouration of the epidermis, loss of skin elasticity and flexibility and crust formation. All treated skin sites appeared normal at the 14-day observation. No corrosive effects were noted.

Conclusion. The test material produced a primary irritation index of 3.0 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme.

Rabbit Number
and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 14

67624Male

2.72

3.09

0.37

67679Male

2.87

3.40

0.53

67680Male

2.54

2.91

0.37


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

Cf = Crust formation

Br = Light brown discolouration of the epidermis

Le = Loss of skin elasticity

Lf = Loss of skin flexibility

Interpretation of results:
GHS criteria not met
Conclusions:
Moderate irritation but the effects recorded were reversible. Not classified.
Executive summary:

Introduction. 

The study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

*       OECD Guidelines for the Testing of Chemicals No. 404 “Acute Dermal Irritation/Corrosion” (adopted)

*        Method B4 Acute Toxicity (Skin Irritation) of Commission Directive 2004/73/EC

Results. 

3-minute and 1-hour semi-occluded applications of the test material to the intact skin of one rabbit produced no corrosive effects.

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced well-defined erythema and very slight oedema. Other skin reactions noted were light brown discolouration of the epidermis, loss of skin elasticity and flexibility and crust formation. All treated skin sites appeared normal at the 14-day observation. 

.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Remarks:
The study has been conducted according to guideline OECD TG 405.
Type of information:
experimental study
Adequacy of study:
key study
Study period:
between 26 August 2008 and 04 September 2008.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Date of inspection: 21/08/2007 Date of signature: 15/10/07
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source:
Harlan UK Limited, Bicester, Oxon, UK

- Age at study initiation:
twelve to twenty weeks old

- Weight at study initiation:
At the start of the study the animals were in the weight range of 2.0 to 3.5 kg

- Housing:
The animals were individually housed in suspended cages

- Diet (e.g. ad libitum):
Free access to food (2030 Teklad Global Rabbit diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK) was allowed throughout the study

- Water (e.g. ad libitum):
Free access to mains drinking water was allowed throughout the study

- Acclimation period:
at least five days


ENVIRONMENTAL CONDITIONS
- Temperature (°C):
The temperature awas set to achieve limits of 17 to 23°C

- Humidity (%):
The relative humidity was set to achieve limits of 30 to 70%

- Air changes (per hr):
The rate of air exchange was at least fifteen changes per hour

- Photoperiod (hrs dark / hrs light):
the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness
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

- Concentration (if solution):
For the purpose of the study the test material was used as supplied.

VEHICLE
Not applicable
Duration of treatment / exposure:
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours
Observation period (in vivo):
Not applicable
Number of animals or in vitro replicates:
Initially, a single rabbit was treated

After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.
Details on study design:
REMOVAL OF TEST SUBSTANCE
Not applicable

SCORING SYSTEM:
The numerical values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae were designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of cornea involved). For each tissue the score was calculated as follows:

Score for conjunctivae = (A + B + C) x 2
Score for iris = D x 5
Score for cornea = (E x F) x 5

Using the numerical data obtained a modified version of the system described by Kay J H and Calandra J C (1962), J. Soc. Cosmet. Chem. 13, 281 289 (see Appendix 3) was used to classify the ocular irritancy potential of the test material. This was achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation were calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled classification of the eye irritancy potential of the test material.

If evidence of irreversible ocular damage is noted, the test material will be classified as corrosive to the eye.

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: 67702 Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
cornea opacity score
Basis:
animal: 67719 Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
cornea opacity score
Basis:
animal: 67720 Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
iris score
Basis:
animal: 67702 Male
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: No effects observed
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
iris score
Basis:
animal: 67719 Male
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: No effects observed
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
iris score
Basis:
animal: 67720 Male
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: No effects observed
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
conjunctivae score
Basis:
animal: 67702 Male
Time point:
24/48/72 h
Score:
0.3
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
conjunctivae score
Basis:
animal: 67719 Male
Time point:
24/48/72 h
Score:
0.3
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
conjunctivae score
Basis:
animal: 67720 Male
Time point:
24/48/72 h
Score:
0.3
Max. score:
3
Reversibility:
fully reversible within: 48hours
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
chemosis score
Basis:
animal: 67702 Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
chemosis score
Basis:
animal: 67719 Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
other: Initial pain reaction = 2
Irritation parameter:
chemosis score
Basis:
animal: 67720 Male
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
other: Initial pain reaction = 2
Irritant / corrosive response data:
Ocular Reactions
Individual and group mean scores for ocular irritation are given in Table 1 and Table 2.

No corneal or iridial effects were noted during the study.

Moderate conjunctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation noted at the 24-hour observation.

All treated eyes appeared normal at the 48-hour observation.
Other effects:
Individual bodyweights and bodyweight changes are given in Table 3.

Two animals showed expected gain in bodyweight during the study while the remaining animal showed no gain in bodyweight.

Table1               IndividualScores and Individual Total Scoresfor Ocular Irritation

Rabbit Number and Sex

67702Male

67719Male

67720Male

IPR= 2

IPR = 2

IPR = 2

Time After Treatment

1

Hour

24

Hours

48

Hours

72

Hours

1

Hour

24

Hours

48

Hours

72

Hours

1

Hour

24

Hours

48

Hours

72

Hours

CORNEA

 

 

 

 

 

 

 

 

 

 

 

 

E = Degree of Opacity

0

0

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

0

0

Score (E x F) x 5

0

0

0

0

0

0

0

0

0

0

0

0

IRIS

 

 

 

 

 

 

 

 

 

 

 

 

D

0

0

0

0

0

0

0

0

0

0

0

0

Score (D x 5)

0

0

0

0

0

0

0

0

0

0

0

0

CONJUNCTIVAE

 

 

 

 

 

 

 

 

 

 

 

 

A = Redness

2

1

0

0

2

1

0

0

2

1

0

0

B = Chemosis

1

0

0

0

1

0

0

0

1

0

0

0

C = Discharge

1

0

0

0

1

0

0

0

1

0

0

0

Score (A + B + C) x 2

8

2

0

0

8

2

0

0

8

2

0

0

Total Score

8

2

0

0

8

2

0

0

8

2

0

0

 


Table2               Individual Total Scores and Group Mean Scoresfor Ocular Irritation

Rabbit Number

and Sex

Individual Total Scores At:

1 Hour

24 Hours

48 Hours

72 Hours

67702Male

8

2

0

0

67719Male

8

2

0

0

67720Male

8

2

0

0

Group Total

24

6

0

0

Group Mean Score

8.0

2.0

0.0

0.0

 


Table3               Individual Bodyweights and Bodyweight Change

Rabbit Number
and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 14

67702Male

3.05

3.09

0.04

67719Male

2.67

2.67

0.00

67720Male

2.63

2.68

0.05


IPR= Initial pain reaction

Interpretation of results:
GHS criteria not met
Conclusions:
Slightly irritating and all recorded effects reversible within 48 hours. Not classified.
Executive summary:

Introduction. The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

* OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted)Method B5 Acute

* Toxicity (Eye Irritation) of Commission Directive 2004/73/EC

Result. A single application of the test material to the non-irrigated eye of three rabbits produced moderate conjunctival irritation. All treated eyes appeared normal at the 48-hour observation.

Conclusion. The test material produced a maximum group mean score of 8.0 and was classified as a minimal irritant (Class 3 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

In a skin irritation/corrosion study, 0.5 ml of Oxooil LS9 was applied under a semi-occluded dressing to the intact shaven dorsal skin of three New Zealand White rabbits for 4 hours, followed by a 14 day observation period. All three rabbits showed well-defined erythema and very slight oedema. Other skin reactions noted were light brown discolouration of the epidermis, loss of skin elasticity and flexibility and crust formation. All treated skin sites appeared normal at the 14-day observation. The mean score for each animal at 24, 48 and 72 hours after removal of the dressing was 2 for erythema and 1 for oedema. No corrosive effects were recorded.

In an eye irritation study, Oxooil LS9 (0.1 ml) was instilled into the right eye of three New Zealand White rabbits. Responses to treatment were assessed using the Draize method 1, 24, 48 and 72 hrs after instillation and again on Days 6, 8, 10, 14, 17 and 21. Reactions to treatment were confined to slight or moderate conjunctival redness, very slight oedema and slight ocular discharge in all three rabbits at the 1and/or 24 hour examinations. The mean scoresfor each animal at the 24, 48 and 72 hour examinations were 0.3 for conjunctival redness and 0 for oedema.


Justification for classification or non-classification

In an acute skin irritation study, the mean score for erythema at the 24, 48 and 72 hour examinations for all three animals (2) was equal to the classification threshold (2) for classification as irritant to skin, according to the general classification and labelling requirements for dangerous substances and preparations (Directive 67-548-EEC). However, the mean score for erythema at the 24, 48 and 72 hour examinations for all three animals (2), is below the classification threshold (2.3) for classification as irritant to skin, according to the requirements of the classification, labelling and packaging (CLP) regulation (EC) No 1272/2008.

In an acute eye irritation study, effects of treatment were confined to transient changes to the conjunctivae in the first 24 hours. Oxooil LS9 is therefore classified as non-irritant to eyes, according to the general classification and labelling requirements for dangerous substances and preparations.