Registration Dossier

- Skin irritation: not irritating, female, New-Zealand White rabbit, OECD TG 404, Schneider 1988

- Eye irritation: not irritating, male, New-Zealand White rabbit, OECD TG 405, Schneider 1988

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 Feb 1988 to 05 Feb 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

May 1981

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

KFM-NZW

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: approx. 12 – 14 weeks old.
- Weight at study initiation: 2250 - 2510 g.
- Housing: Rabbits were individually housed in metal wire rabbit cages.
- Diet: standard rabbit pellet, ad libitum
- Water: fresh water, ad libitum. The quality of the drinking water was according to the specifications of the "Schweizerisches Lebensmittelbuch".
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g of the test substance

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

STUDY DESIGN
- The test was performed on 3 female rabbits, checked for normal skin conditions.
- The animals were checked daily for systemic symptoms and mortality.
- The body weight was recorded at start and on day 3 of the test.

TEST SITE
- Area of exposure: An area of at least 36 cm2 was shaved on both flanks of the animals approximately 24 hours before treatment.
- Type of wrap: A gauze patch (20 cm2) was applied to the flank. A control gauze patch was applied to the contralateral flank. Both gauze patches were moistened before application with distilled water containing 0.5 % carboxymethylcellulose and 0.1 % polysorbate 80. The patches were loosely covered with an aluminium foil (36 cm2) and held in place by an adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
Approximately 1, 24, 48 and 72 hours after the end of exposure

SCORING SYSTEM:
- Method of calculation: following the OECD TG 404 system. For criteria, see 'Any other information on materials and methods incl. tables'.

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Two animals exhibited slight erythema reactions (score 1) one hour after treatment with test substance. Scores of erythema and oedema for the recordings 24 to 72 hours in all 3 animals were zero (considered to be non-irritating).

Other effects:

BODY WEIGHT
The body weights of all rabbits were considered to be within the normal range of variability.

DURATION OF THE IN-LIFE PHASE
Because no reactions were observed at 24 hours to 72 hours after removing the bandages, the test was ended after the 72 hours evaluation.

Table 2 Individual skin reaction score

	E R Y T H E M A			E D E M A
animal	#1	#2	#3	#1	#2	#3
animal	CF/TF	CF/TF	CF/TF	CF/TF	CF/TF	CF/TF
after 1 hr	0/0	0/1	0/1	0/0	0/0	0/0
after 24 hrs	0/0	0/0	0/0	0/0	0/0	0/0
after 48 hrs	0/0	0/0	0/0	0/0	0/0	0/0
after 72 hrs	0/0	0/0	0/0	0/0	0/0	0/0
mean 24-72 hrs.	0/0	0/0	0/0	0/0	0/0	0/0

CF = control flank TF = test flank

M = male F = female

Table 3 Body Weights

Animal	#1	#2	#3
At start of test	2250	2510	2480
After 3 days (end)	2310	2570	2490

Interpretation of results:

GHS criteria not met

Conclusions:

Because the mean values of the recordings 24 to 72 hours after application were scored zero, the test substance can be classified as non-irritant.

Executive summary:

The primary skin irritation potential of the test substance was investigated according to the GLP compliant OECD TG 404. An amount of 0.5 g unchanged test material was applied under occlusive conditions for 4 hours to the clipped backs of 3 females albino New Zealand White rabbits (KFM-NZW), using a gauze patch of 20 cm2. A control gauze patch was applied to the contralateral flank. Both gauze patches were moistened before application with distilled water containing 0.5 % carboxymethylcellulose and 0.1 % polysorbate 80. The patches were loosely covered with an aluminium foil (36 cm2) and held in place for 4 hours by an adhesive tape. Scoring was performed at 1, 4, 24, 48 and 72 hours and after 3 days of application following OECD TG 404.

Two animals exhibited slight erythema reactions (score 1) one hour after treatment with test substance. Since no reactions were observed at 24 hours to 72 hours after removing the bandages, the test was terminated after the 72 hours evaluation. Scores of erythema and oedema for the recordings 24 to 72 hours in all 3 animals were zero. Under the conditions of this study, the test substance was considered not to be irritating to the skin of rabbits.

Endpoint conclusion:

no adverse effect observed (not irritating)

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 Feb 1998 to 12 Feb 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Feb 1987

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

KFM-NZW

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: 12 - 14 weeks
- Weight at study initiation: 2410 - 2640 g
- Housing: The animals were housed individually in metal cages.
- Diet: standard rabbit pellet, ad libitum (analysed for nutritive ingredients and contaminants)
- Water: Ad libitum.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 09 Feb 1998 To 12 Feb 1988

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- 0.1 mL (70 mg)

Duration of treatment / exposure:

Single treatment

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 males

Details on study design:

INSTILLATION
The test substance was placed into the conjunctival sac of the left eye of each animal, after gently pulling away the lower lid from the eyeball. The lids were then held together for about one second in order to prevent loss of the test material. The right eye remained untreated and served as a control.

OCULAR SCORING
The ocular reactions were evaluated 1, 24, 48, and 72 hours after the instillation of the test substance according to the OECD scoring system (see Table 1 at ‘Any other information on methods and materials incl. tables’). A slit-lamp was used to facilitate the evaluation.
The irritant/corrosive potency of test substance was classified according to the EC Commission Directive No. 83/467, 1983. Ocular lesions which occur within 72 hours after exposure and which persist for at least 24 hours and correspond to the following values determined on the rabbit according to the eye irritation method:
- The mean value of the scores for each type of lesion, calculated over all the animals tested, is one of the following:
(1) cornea opacity two or more
(2) iris lesion one or more
(3) redness of conjunctivae 2.5 or more
(4) oedema of conjunctivae
(5) (chemosis) two or more
- or, in the case where the Annex V test has been completed using three animals, either cornea opacity, iris lesion, redness of conjunctivae or oedema of conjunctivae (chemosis) equivalent to a mean value such as is quoted above, but calculated for each animal separately, has been observed in two or more animals.
In both cases all scores at each of the reading times (24, 48, and 72 hours) and for an effect should be used in calculating the respective means values.

CAGE SIDE OBERVATIONS
The animals were checked daily for systemic symptoms and mortality..

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The test substance induced reactions in the cornea (score 1), iris (score 1) and conjunctiva (redness score 1 and/or chemosis score 1) in all 3 animals at the 1-hour reading. At the 24-hour reading, only 1 animal still exhibited redness of the conjunctiva (score 1). The eye reactions observed were reversible until the end of the observation period on day 3.

Other effects:

BODY WEIGHT
The body weights of all rabbits were considered to be within the normal range of variability.

Table 2 Scores of eye irritation after instillation of test substance

	Cornea	Iris	Conjunctiva
	Cornea	Iris	redness	chemosis
score (3 animals investigated)	0 -4	0 - 2	0 - 3	0 - 4
60 min	1/1/1	1/1/1	1/1/1	1/0/1
24 hours	0/0/0	0/0/0	0/0/1	0/0/0
48 hours	0/0/0	0/0/0	0/0/0	0/0/0
72 hours	0/0/0	0/0/0	0/0/0	0/0/0
average 24h, 48h, 72h	0/0/0	0/0/0	0/0/0.33	0/0/0
Reversibility*	c	c	c	c
time for reversion	24 h	24 h	48 h	24 h

* c : completely reversible

Table 3 Body weight

Animal no.	#1	#2	#3
At start of test	2410	2630	2640
After 3 days (end)	2460	2710	2670

Interpretation of results:

GHS criteria not met

Conclusions:

Based on this study, it was concluded that test substance was considered to be not irritating to the eyes of rabbits.

Executive summary:

In a primary eye irritation study performed in accordance with OECD TG 405 following GLP principles. An amount of 0.1 mL (70 mg) of the undiluted test substance into the conjunctival sac of the left eye of each the 3 males albino New Zealand White rabbits (KFM-NZW), after gently pulling away the lower lid from the eyeball. The lids were then held together for about one second in order to prevent loss of the test material. The right eye remained untreated and served as a control. The animals were checked daily for systemic symptoms and mortality. The ocular reactions were evaluated 1, 24, 48, and 72 hours after the instillation of test article. A slit-lamp was used to facilitate the evaluation. The animals were checked daily for systemic symptoms and mortality.

The test substance induced reactions in the cornea (score 1), iris (score 1) and conjunctiva (redness score 1 and/or chemosis score 1) in all 3 animals at the 1-hour reading. At the 24-hour reading, only 1 animal still exhibited redness of the conjunctiva (score 1). The eye reactions observed were reversible until the end of the observation period on day 3. The body weights of all rabbits were considered to be within the normal range of variability.

Under the conditions of this study, the test substance was considered to be not irritating to the eyes of rabbits.

Endpoint conclusion:

no adverse effect observed (not irritating)

Endpoint conclusion:

no study available

Skin irritation in rabbits, Schneider 1988

The primary skin irritation potential of the test substance was investigated according to the GLP compliant OECD TG 404. An amount of 0.5 g unchanged test material was applied under occlusive conditions for 4 hours to the clipped backs of 3 females albino New Zealand White rabbits (KFM-NZW), using a gauze patch of 20 cm2. A control gauze patch was applied to the contralateral flank. Both gauze patches were moistened before application with distilled water containing 0.5 % carboxymethylcellulose and 0.1 % polysorbate 80. The patches were loosely covered with an aluminium foil (36 cm2) and held in place for 4 hours by an adhesive tape. Scoring was performed at 1, 4, 24, 48 and 72 hours and after 3 days of application following OECD TG 404.

Two animals exhibited slight erythema reactions (score 1) one hour after treatment with test substance. Since no reactions were observed at 24 hours to 72 hours after removing the bandages, the test was terminated after the 72 hours evaluation. Scores of erythema and oedema for the recordings 24 to 72 hours in all 3 animals were zero. Under the conditions of this study, the test substance was considered not to be irritating to the skin of rabbits.

Eye irritation in rabbits, Schneider 1988

In a primary eye irritation study performed in accordance with OECD TG 405 following GLP principles. An amount of 0.1 mL (70 mg) of the undiluted test substance into the conjunctival sac of the left eye of each the 3 males albino New Zealand White rabbits (KFM-NZW), after gently pulling away the lower lid from the eyeball. The lids were then held together for about one second in order to prevent loss of the test material. The right eye remained untreated and served as a control. The animals were checked daily for systemic symptoms and mortality. The ocular reactions were evaluated 1, 24, 48, and 72 hours after the instillation of test article. A slit-lamp was used to facilitate the evaluation. The animals were checked daily for systemic symptoms and mortality.

The test substance induced reactions in the cornea (score 1), iris (score 1) and conjunctiva (redness score 1 and/or chemosis score 1) in all 3 animals at the 1-hour reading. At the 24-hour reading, only 1 animal still exhibited redness of the conjunctiva (score 1). The eye reactions observed were reversible until the end of the observation period on day 3. The body weights of all rabbits were considered to be within the normal range of variability.

Under the conditions of this study, the test substance was considered to be not irritating to the eyes of rabbits.

Based on the available information, classification for skin and eye irritation is not warranted in accordance with EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation No. (EC) 1272/2008.