Alkali salt of (sulfonato oxyethane sulfonyl) arylamino triazinylamino sulfonato (sulfonato arylazo) aryl sulfonate

Endpoint summary

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:

From June 27, 2006 to June 30, 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD Guideline 404, EU Method B.4 and EPA OPPTS 870.2500 Method, in compliance with GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

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:

TEST ANIMALS
- Source: Ferenc Sandor breeder, 2173 Kartal, Csaszar ut 135, Hungary
- Date of receipt: June 20, 2006
- Age at study initiation: 10 weeks old, adult albino rabbit
- Body weight range at treatment: 2,553-2,687g (i.e., at the beginning of the study), 2,668-2,858 g (i.e., at the end of the study)
- Sex: Male.
- Housing: Animals were housed individually in metal cages
- Diet: Purina Base – Lap gr. diet for rabbit produced by Agribrands Europe Hungary PLC, H-5300 Karcag, Madarasi road, Hungary, ad libitum
- Water: Tap water from self- service water system, ad libitum
- Animal health: Only animals in acceptable health condition were used for the test.
- Acclimation period: 7 d
- Animal identification: The individual identification was performed by numbered ear tag. The cages were marked with individual identity cards with information about study code, sex, dose group, cage number and individual animal number.

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3°C
- Relative humidity: 30-70%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: June 27, 2006 to June 30, 2006

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated skin of each animal

Amount / concentration applied:

500 mg

Duration of treatment / exposure:

4 h

Observation period:

1, 24, 48 and 72 h after the patch removal

Number of animals:

3

Details on study design:

ADMINISTRATION OF THE TEST SUBSTANCE

Dosage
The test substance was used in pure state, in a single dose. 0.5 g of test substance was applied to the test site. The test substance was moistened sufficiently with water to ensure good contact with the skin. Untreated skin of each animal served as control.

TEST PROCEDURE
Patch testing was used to detect primary irritating effects of the test substance. Three male animals in acceptable health condition were selected for this test. Each animal was examined 24 h before starting the test. Approximately 24 h prior to the test the hair was clipped from the back and flanks of the animals.
The test substance was applied to an approximately 6 cm² area of intact skin. Sterile gauze pads (i.e., 5x5 cm) were placed on the skin of rabbits. These gauze patches were kept in contact with the skin by a patch with a surrounding adhesive hypoallergenic plaster. The entire trunks of the animals were wrapped with plastic wrap for 4 h. In the first step an initial test was performed using one animal. 1 h after application of the test substance to the sentinel animal, the application site was examined. Severe irritation or corrosive effect could not be found in the initial test, therefore the bandage was replaced and the exposure continued for a further 3 h (i.e., a total 4 h exposure). Two additional animals were involved for this study.

Duration of Exposure
Duration of exposure: 4 h. After the treatment period the rest of the test substance was removed with body temperature water.

OBSERVATIONS AND SCORING

Clinical Observations
Animals were examined for signs of erythema and oedema, and the responses scored at 60 minutes and then at 24, 48 and 72 h after the patch removal. At the end of the observation period euthanasia of the animal was by intramuscular injections of Ketamin and Xylazin followed by i.v. Euthanyl anaesthesia.

Scoring and Assessment of Local Reactions
The dermal irritation scores were evaluated according to the scoring system by Draize (1959). The animals were observed for 72 h, the duration of the study was sufficient to evaluate fully the reversibility or irreversibility of the effects observed.

SCORING SYSTEM:
Erythema and eschar formation
No erythema........................................................................................................................0
Very slight erythema (barely perceptible) ......................................................................1
Well-defined erythema ......................................................................................................2
Moderate to severe erythema............................................................................................3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) .......4

Oedema
No oedema..............................................................................................................................0
Very slight oedema (barely perceptible).............................................................................1
Slight oedema (edges of area well defined by definite raising).......................................2
Moderate oedema (raised approximately 1 mm) ..............................................................3
Severe oedema (raised more than 1 mm and extending beyond area of exposure)...4

Measurement of Body Weight
Body weights were recorded at the beginning and at the end of experiment.

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

- 1 h after the patch removal in two animals very slight oedema appeared on the treated skin surface. The erythema could not be evaluated in consequence of covered up colour of the test substance, but the possibility of the severe erythema and corrosion were precluded.
- 24, 48 and 72 h after the patch removal all animals were symptom-free, so the study was terminated 72 h after the patch removal.
- During the study the control area was symptom-free in all animals.

Other effects:

- General state and the behaviour of animals were normal throughout the study period.
- There were no notable body weight changes during the contact and observation period.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions, the test substance was not considered to be irritating to skin.

Executive summary:

A study was performed to assess the skin irritation potential of the test substance in male New Zealand White rabbits according to OECD Guideline 404, EPA OPPTS 870.2500 and EU Method B.4, in compliance with GLP.

500 mg of test substance moistened sufficiently with water was applied to an approximately 6 cm² area of intact skin. Sterile gauze pads (i.e., 5x5 cm) were placed on the skin of rabbits surrounded by an adhesive hypoallergenic plaster. The entire trunks of the animals were wrapped with plastic wrap for 4 h. After the exposure period, all remnants of test substance were carefully removed from the skin with body temperature water. Animals were scored for irritation reactions after 1, 24, 48 and 72 h.

Very slight oedema was observed in two animals at 1 h after the patch removal. Although, the erythema could not be evaluated in consequence of covered up colour of the test substance, the possibility of the severe erythema and corrosion was excluded. All animal were symptom free at 24, 48 and 72 h. Further, overall mean scores for dermal irritation were calculated to be 0.00 for erythema and 0.00 for edema formation. In addition, no clinical signs of systemic toxicity and no notable body weight changes were observed during the study period.

 

Under the test conditions, the test substance was not considered to be irritating to skin.

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:

From July 07, 2006 to July 10, 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD Guideline 405, EPA OPPTS 870.2400 and EU Method B.5, in compliance with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: 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:

TEST ANIMALS
- Source: Ferenc Sandor breeder, 2173 Kartal, Csaszar ut 135, Hungary
- Date of receipt: June 27, 2006
- Age at study initiation: 10 weeks old, adult albino rabbit
- Body weight range at treatment: 2,706-2,838 g (i.e., at the beginning of the study), 2,746-2,908 g (i.e., at the end of the study)
- Sex: Male.
- Housing: Animals were housed individually in metal cages
- Diet: Purina Base – Lap gr. diet for rabbit produced by Agribrands Europe Hungary PLC, H-5300 Karcag, Madarasi road, Hungary, ad libitum
- Water: Tap water from automatic self-supplying watering system, ad libitum
- Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined 24 h prior to the start of study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.
- Acclimation period: 10 d
- Animal identification: The individual identification was performed by numbered ear tag. The cages were marked with individual identity cards with information about study code, sex, dose group, cage number and individual animal number.

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3°C
- Relative humidity: 30-70%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: July 07, 2006 to July 10, 2006

Vehicle:

unchanged (no vehicle)

Controls:

other: contralateral eye

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 24, 48 and 72 h following administration of the test substance.

Number of animals or in vitro replicates:

3 males

Details on study design:

Test Procedure
Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition, any with ocular lesions were rejected.
In the first step an initial test was performed using one animal. The test substance was instilled into the conjunctival sac of the left eye. The eyelids were held closed gently for several seconds to prevent the loss of the test substance. The contralateral eye served as control. Immediately after the administration of the test substance, an assessment of the initial pain reaction was made.

After consideration of the ocular responses produced in the first animal, two additional animals were treated. Before the administration the treated eyes of animals were not anaesthetised, because the score of initial pain reaction in the first animal indicated practically no initial pain.

Duration of Exposure
The eyes of the test animals were not washed out after the application of test substance.

OBSERVATIONS AND SCORING

Clinical Observations
The eyes were examined at 1, 24, 48 and 72 h after the treatment. The duration of the observation period was sufficient for the statement of reversibility or irreversibility of changes. Any clinical signs of toxicity or signs of ill-health during the study were recorded. At the end of the observation period all animals were sacrificed by intramuscular injections of Ketamin and Xylazin followed by i.v. Euthanyl anaesthesia.

Scoring and Assessment of Local Reaction
The eye irritation scores were evaluated according to the scoring system by Draize (1959) and OECD 405 (24 April 2002).

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

(mean score)

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

2

Irritant / corrosive response data:

- 1h after treatment in all animals the vessels were more diffuse, with obvious conjunctival swelling with partial eversion of the lids and the discharge with moistening of the lids and hairs on considerable area around the eye was found.
- Corneal and iris alterations were not recorded throughout the study period.
- 24 h after treatment in all animals displayed some hyperaemic blood vessels. In two animals the discharge from the eye was greater than normal. In one animal some swelling above normal was observed at this time.
- 48 h after treatment every animal was symptom-free.
- 72 h after treatment the study was terminated, since no primary irritation symptoms occurred.
- During the study the control eyes of animals were symptom-free.

Other effects:

- General state and the behaviour of animals were normal throughout the study period.
- There were no notable body weight changes during the contact and observation period.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions, the test substance was not considered to be irritating to eyes.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in male New Zealand White rabbits according to OECD Guideline 405, EPA OPPTS 870.2400 and EU Method B.5, in compliance with GLP.

A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of each animal. The treated eyes were not washed out after admnistration. The untreated eyes served in each case as a control.

Moderate redness, chemosis and severely increased discharge were observed 1 h after the single application. All animals were free of irritation symptoms after 48 h. Further, no corneal and iris alterations were observed throughout the study period. In addition, no clinical signs of systemic toxicity and no notable body weight changes were seen during the study period.

Under the test conditions, the test substance was not considered to be irritating to eyes. 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

A study was performed to assess the skin irritation potential of the test substance in male New Zealand White rabbits according to OECD Guideline 404, EPA OPPTS 870.2500 and EU Method B.4, in compliance with GLP.

500 mg of test substance moistened sufficiently with water was applied to approximately 6 cm² area of intact skin. Sterile gauze pads ( 5x5 cm) were placed on the skin of rabbits surrounded by an adhesive hypoallergenic plaster. The entire trunks of the animals were wrapped with plastic wrap for 4 h. After the exposure period, all remnants of test substance were carefully removed with body temperature water. Animals were scored for irritation reactions after 1, 24, 48 and 72 h. Very slight oedema was observed in two animals at 1 h after the patch removal. Although, the erythema could not be evaluated in consequence of covered up colour of the test substance, the possibility of severe erythema and corrosion was excluded. All animal were symptom free at 24, 48 and 72 h. Further, overall mean scores for dermal irritation were calculated to be 0.00 for erythema and 0.00 for edema formation. In addition, no clinical signs of systemic toxicity and no notable body weight changes were observed during the study period. Under the test conditions, the test substance was considered not to be irritating to skin (Stahl, 2006a).

A study was conducted to assess the eye irritancy potential of the test substance in male New Zealand White rabbits according to OECD Guideline 405, EPA OPPTS 870.2400 and EU Method B.5, in compliance with GLP.

A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of each animal. The treated eyes were not washed out after administration. The untreated eyes served in each case as a control. Moderate redness, chemosis and severely increased discharge were observed 1 h after application. All animals were free of irritation symptoms after 48 h. Further, no corneal and iris alterations were observed throughout the study period. In addition, no clinical signs of systemic toxicity and no notable body weight changes were seen during the study period. Under the test conditions, the test substance was not considered to be irritating to eyes (Stahl, 2006b).

Justification for selection of skin irritation / corrosion endpoint:
Guideline-compliant study conducted according to GLP.

Justification for selection of eye irritation endpoint:
Guideline-compliant study conducted according to GLP.

Effects on eye irritation: slightly irritating