R-AMINR

Endpoint summary

Description of key information

Reliable OECD guideline studies 404/405 are available for the test substance assessing the potential for skin and eye irritation. Both studies were conducted in accordance with GLP.

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Species:

rabbit

Strain:

New Zealand White

Remarks:

Crl:NZW/Kbl(BR)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Ltd, Manston.
- Age at study initiation: Fourteen to sixteen weeks old
- Weight at study initiation: 2.7 to 2.94 Kg
- Housing: Single floow pens with a minimum area of 0.6 sq m.
- Diet (e.g. ad libitum): SQC standard rabbit diet (pelleted) was freely available at all times.
- Water (e.g. ad libitum): Mains water provided ad libitum via water bottles.
- Acclimation period: At least 1 month

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 to 22°C
- Humidity (%): 40 to 80%
- Air changes (per hr): At least 10
- Photoperiod (hrs dark / hrs light): Twelve hours light/dark cycle

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 mg spread uniformly over the designated area.

Duration of treatment / exposure:

4 hours

Observation period:

Not less than 3 days

Number of animals:

3 males (1 sentinel followed by 2 additional animals)

Details on study design:

TEST SITE
- Area of exposure: 30 x 20 mm
- Type of wrap if used: Area covered by a dense gauze patch which in trun was covered by a larger gauze patch and an open weave elasticated adhesive bandage firmly wrapped around the torso.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated areas of the skin were swabbed with moist cotton wool.
- Time after start of exposure: Straight away after the 4 hour exposure period.

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) 1, 24, 48 and 72 hours after patch removal

SCORING SYSTEM:
- Method of calculation: Standard as per OECD guidelines.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #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

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

There were no dermal changes or irritation reactions at sites of application of the test substance.

Other effects:

There were no signs of toxicity or ill health in any rabbit during the observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

A single semi-occluded application of the test substance to intact rabbit skin for 4 hours did not elicit any dermal changes or irritation reactions.

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

Crl:NZW/Kbl(BR)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Ltd.
- Age at study initiation: Sixteen to nineteen weeks old
- Weight at study initiation: 3.16 to 3.57 Kg
- Housing: Single floor pens accomodating 1 rabbit with a minium floor area of 0.6 sq m.
- Diet (e.g. ad libitum): SQC standard rabbit diet (pelleted) was freely available at all times.
- Water (e.g. ad libitum): Mains water was provided ad libitum.
- Acclimation period: At least 1 month.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 to 22°C
- Humidity (%): 40 to 80% RH
- Air changes (per hr): At least 10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle

Vehicle:

water

Remarks:

Dispersed in distilled water

Controls:

not required

Amount / concentration applied:

The test article was dispersed in distilled water and the pH of the 50% preparation was confirmed not to exceed 11.5 or to be less than 2 before dosing commenced. The test article was administered as a known weight (67 mg) with a compacted volume of 0.1 mL. Doses were expressed in terms of test article received and all doses were used on the day or preparation.

Duration of treatment / exposure:

A singular ocular instillation

Observation period (in vivo):

Not less than 3 days

Number of animals or in vitro replicates:

3 rabbits (one sentinel followed by a further two exposures)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
Not performed

SCORING SYSTEM:
Standard as per OECD 405 guideline. The 'initial sting' response was assessed and recorded immediately after instillation of the test article. Treated animals were inspected for indications of distress or unacceptable ocular changes frequently during Day 1 (the day of dosing) and at least daily until completion of the observation period. Ocular changes were assessed and recorded immediately after treatment, one half hour, one and four hours after treatment on Day 1, and 24, 48 and 72 hours after treatment.

TOOL USED TO ASSESS SCORE: The treated eyes were examined visually or with the aid of a pencil-beam torch, biomicroscope or other device to illuminate or magnify the eye during examination. At examinations carried out 24 hours after treatment the cornea was subject to application of 2% aqueous fluorescein solution followed by irrigation with water for irrigation and illumination by an ultraviolet source. This facilitated detection of damage to the corneal epithelium as indicated by absorption of the fluorescing dye into the epithelium or stroma. The application of fluorescein was repeated in two rabbits 48 hours after treatment.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

48 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

mean

Time point:

48 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Instillation of the substance caused practically no initial sting response or a slight initial sting response. All rabbits developed a crimson-red appearance of the conjunctivae shortly after treatment. This was associated with slight chemosis in two rabbits and with a slight ocular discharge in one of the same animals. No conjunctival reactions of greater intensity developed later in the study but resolution of conjunctival irritation remained incomplete until three days after treatment.

The cornea of one rabbit showed an area (approximately 2 x 5 mm) of slight opacity that was also permeable to applied fluorescein on the day after ocular instillation of the test article. This corneal change resolved within two days of treatment. The iris remained overtly unaffected by the test substance.

Other effects:

No observations indicative of systemic toxicity or ill health were noted for any rabbits during the course of the study.

Group mean values for ocular reactions

Time of observation	Ocular reaction
	Corneal opacity	Iris	Conjunctival redness	Chemosis
1 hour	0	0	1	0
4 hour	0	0	1.3	0.7
24 hour	0.3	0	0.7	0.3
48 hour	0	0	0.7	0.3
72 hour	0	0	0	0
Mean of 24, 48, and 72 hour scores	0.1	0	0.4	0.2

Interpretation of results:

GHS criteria not met

Conclusions:

Single instillation of the test substance to the conjunctival sac of three rabbits caused reversible changes of the conjunctivae in all animals and a transient and minor opacity of the cornea of one animal. However, the effects observed did not merit classification under UN GHS or the CLP Regulation (EC No. 1272/2008).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

No skin effects or signs of irritation were observed in the available in vivo skin irritation study and accordingly the criteria for classification under the CLP Regulation (EC 1272/2008, as amended) is not met. Similarly effects were observed in the available in vivo eye irritation study did not met the CLP classification criteria based on the mean values at 24, 48 and 72 hours.