2-(4,6-bis(2,4-dimethylphenyl)-1,3,5-triazin-2-yl)-5-(3-((2-ethylhexyl)oxy)-2-hydroxypropoxy)phenol

Endpoint summary

Description of key information

To evaluate the irritation potential of the test article, skin or eyes of New Zealand rabbits were treated with the test substance and reactions were observed and scored up to 72h (according to OECD guidelines 404 and 405). Application onto skin did not cause irritation. In contact with eyes, slight reactions were observed which were reversible after 24h. Therefore, the test substance is considered to be non-irritating to skin or eyes.

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:

May 1995

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:

17 Jul 1992

Deviations:

yes

Remarks:

The rabbits used in the study were slightly lighter and younger than proposed

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Version / remarks:

31 Jul 1992

Deviations:

yes

Remarks:

The rabbits used in the study were slightly lighter and younger than proposed

Qualifier:

according to guideline

Guideline:

EPA OTS 798.4470 (Acute Dermal Irritation)

Version / remarks:

1 Jul 1993

Deviations:

yes

Remarks:

The rabbits used in the study were slightly lighter and younger than proposed

Principles of method if other than guideline:

The rabbits used in the study were slightly lighter and younger than proposed in the protocol

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: Z2651-01
- Expiration date of the lot/batch: Feb 1997
- Purity test date: ˃ 95%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (ca. 20°C) away from sunlight
- Stability under test conditions: stable at storage conditions
- Solubility and stability of the test substance in the solvent/vehicle (in bidistilled water): not specified; excluded from the Statement of Compliance

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was moistened with bidistilled water before application

OTHER SPECIFICS:
- pH: not assessed (insoluble in water)
- Description: yellowish solid

Species:

rabbit

Strain:

New Zealand White

Remarks:

CRL:KBL(NZW)BR

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Stolzenseeweg 32-36 0-88353 Kisslegg
- Age at study initiation: 10-12 weeks
- Weight at study initiation: males: 1,9 kg, females: 2 kg
- Weight at start of treatment: males: 2,1 kg, females: 2,1 kg
- Identification: By unique cage number and corresponding ear tags
- Housing: Individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Kliba 341, Batch 89/95 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum.
- Water (e.g. ad libitum): Community tap water from Itingen, ad libitum.
- Acclimation period: Four days under test conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23 °C
- Humidity (%): humidity between 40-70%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light (approx. 100 Lux) / 12 hours dark (light period between 6.00 a.m. to 6.00 p.m.), music during the light period.

IN-LIFE DATES:
Treatment May 15, 1995
Observation May 15 -18, 1995
Termination May 18, 1995

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g/animal (left side only)

Duration of treatment / exposure:

4h

Observation period:

72h

Number of animals:

1 male, 2 female

Details on study design:

Before treatment:
Approximately 24 hours before treatment, the dorsal fur was clipped with an electric clipper, exposing an area of approximately 100 square centimeters (10 cm X 10 cm). The skin of the animals was examined once, 1 day prior to test article administration. Only those animals with no signs of skin injury or irritation were used in the test.

TEST SITE
- Area of exposure: dorsal / back
- coverage: 6 m2
- Type of wrap if used: patch of surgical gauze

REMOVAL OF TEST SUBSTANCE
- Washing (if done): skin was flushed with lukewarm tap water to clean the application site so that the reactions (erythema) were clearly visible at that time.
- Time after start of exposure: 4h

OBSERVATIONS
Viability/Mortality and Clinical Signs: Daily during the observation period.
Body Weights: At the start of acclimatization, on day 1 of test (application day) and at termination of observation.

SCORING SYSTEM:

The skin reaction was assessed at approximately 1, 24, 48 and 72 hours after the removal of the dressing, gauze patch and test article.
The possible corrosive properties of the test article and the staining of the treated skin area related to the test article were described and recorded.

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
Maximum possible score 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 mm) 3
Severe edema (raised more than 1 mm and
extending beyond area of exposure) 4
Maximum possible score 4

Maximum cumulative score: 8

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No corrosive effects were noted on the treated skin of any animal at any measuring

Table 1: Results skin irritation/skin corrosion: Mean values of the scores for each type of lesion, calculated for each animal separately

	Mean
Animal No.	Erythema	Edema
31, male	0.00	0.00
32, female	0.00	0.00
33, female	0.00	0.00

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results and on the referred classification criteria (according to EEC Commission Directive 93/21/EEC, April 27, 1993) the test substance is classified as "not irritating" to rabbit skin.

Executive summary:

The primary skin irritation potential of the test material was investigated by topical application of 0.5 g onto 6 cm2 intact dorsal skin of each of three young adult New Zealand rabbits. The scoring of skin reactions was performed 1, 24, 48 and 72 hours after removal of the dressing. The scores of each animal at the following reading times (24, 48, 72 hours) was used in calculating the respective mean values for each type of lesion. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 hours and then divided by the number of figures. Under the conditions of this experiment, the test substance was found to cause a primary irritation score of 0.00 (max. 8.0) when applied to healthy intact rabbit skin. Local signs (mean values from 24 to 72 hours) consisted of grade 0.00 erythema and grade 0.00 edema. The test article caused no staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval.

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:

May 1995

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:

24 Feb 1987

Deviations:

yes

Remarks:

The rabbits used in the study were slightly lighter and younger than proposed

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

31 Jul 1992

Deviations:

yes

Remarks:

The rabbits used in the study were slightly lighter and younger than proposed

Qualifier:

according to guideline

Guideline:

EPA OTS 798.4500 (Acute Eye Irritation)

Version / remarks:

1 Jul 1993

Deviations:

yes

Remarks:

The rabbits used in the study were slightly lighter and younger than proposed

Principles of method if other than guideline:

The rabbits used in the study were slightly lighter and younger than proposed in the protocol

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: Z2651-01
- Expiration date of the lot/batch: Feb 1997
- Purity: ˃ 95%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (ca. 20°C) away from sunlight
- Stability under test conditions: stable at storage conditions

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was applied as delivered by the sponsor, with the exception that it was ground in a mortar and pestle before administration.

OTHER SPECIFICS:
- pH: not assessed (insoluble)
- Description: yellowish solid

Species:

rabbit

Strain:

New Zealand White

Remarks:

CRL:KBL(NZW)BR

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Stolzenseeweg 32-36 0-88353 Kisslegg
- Age at study initiation: 11-12 weeks
- Weight at start of acclimatization: 1,9 kg
- Weight at start of treatment: male: 2.2kg female: 2.0-2.1 kg
- Identification: By unique cage number and corresponding ear number
- Housing: Individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Kliba 341, Batch 89/95 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum.
- Water (e.g. ad libitum): Community tap water from Itingen, ad libitum.
- Acclimation period: Five days under laboratory test conditions after health examination. Only animals without any visual signs of illness were used for the study, May 11 - 15, 1995

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23 °C
- Humidity (%): humidity between 40-70%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light (approx. 100 Lux) / 12 hours dark (light period between 6.00 a.m. to 6.00 p.m.), music during the light period.

IN-LIFE DATES:
Treatment May 16, 1995
Observation May 16 -19, 1995
Termination May 19, 1995

Vehicle:

unchanged (no vehicle)

Controls:

not required

yes, concurrent no treatment

Amount / concentration applied:

0.1 g / animal (left eye only)

Observation period (in vivo):

72h

Number of animals or in vitro replicates:

1 male, 2 female

Details on study design:

The eyes of the animals were examined for intactness once, one day prior to test article administration. Only those animals with no signs of ocular injury or irritation were used on the test.
On test day 1, 0.1 g of the test article (undiluted) was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test article. The right eye remained untreated and served as the reference control.

SCORING SYSTEM:
EVALUATION OF EYE REACTIONS
CORNEAL IRRITATION
Opacity: degree of density (densest area used for assessment)
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacrous area, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity 4

IRIDIC IRRITATION
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcomeal hyperemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, hemorrhage, gross destruction (any or al 1 of these) 2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris).
Blood vessels normal 0
Some blood vessels definitely hyperemic (injected) 1
Diffuse, crimson color, individual vessels not easily discernible 2
Diffuse beefy red 3

Chemosis: lids and/or nictitating membranes
No swelling 0
Any swelling above normal (includes nictitating membranes) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

The maximum scores are:
Cornea 4
Iris 2
Conjunctival Redness 3
Conjunctival Chemosis : 4

Maximum attainable score 13

TOOL USED TO ASSESS SCORE: Eye examinations were made with a Varta Cliptrix diagnostic-lamp (A. Riegger, Basel / Switzerland).

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24, 48 and 72h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48 and 72h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24, 48 and 72h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 24h

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48 and 72h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 24h

Irritant / corrosive response data:

VIABILITY / MORTALITY AND CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.

IRRITATION
The test substance showed a primary irritation score of: 0.67, when applied to the conjunctival sac of the rabbit eye.

COLORATION
No staining of the cornea, sclera or conjunctivae by the test article was observed.

CORROSION
No corrosion of the cornea was observed at any of the reading times.

BODY WEIGHTS
The body weight of the animals was within the normal range of variability.

Table 1: Results Eye irritation / Eye corrosion: Mean values of the scores for each type of lesion, calculated for each animal separately

	Mean values 24 – 72 hours
Animal No.	Cornea Opacity	Iris	Conjunctivae
			Redness	Chemosis
34, male	0	0	0	0
35, female	0	0	0.67	0.67
36. female	0	0	0.33	0.33

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results and according to classification criteria of EEC Commission Directive 93/21/EEC (April 27, 1993), the test article is classified as "not irritating" to the rabbit eye.

Executive summary:

The primary irritation potential of the test article was investigated by instillation of 0.1 g into one eye of each of three young adult New Zealand rabbits. The treated eyes were not rinsed after application. Scoring of irritation effects were performed 1, 24, 48 and 72 hours after test article application. The scores of each animal at the following reading times (24, 48 and 72 hours) were used in calculating the respective mean values for each type of lesion. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 hours and then dividing the resulting total by the number of figures. It was found to be 0.67 (max. 13), when applied to the conjunctival sac of the rabbit eye. No staining of the cornea, sclera or conjunctivae of the treated eyes by the test article was observed. No corrosion was observed at any of the measuring intervals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

Primary skin irritation properties of the test material were investigated by topical application of 0.5 g onto 6 cm2 intact dorsal skin of each of three young adult rabbits (according to OECD guideline 404). The scoring of skin reactions was performed 1, 24, 48 and 72 hours after removal of the dressing. Under the conditions of this experiment, the test substance did not cause irritation, erythema or oedema when applied to healthy intact rabbit skin. Therefore, the test substance is considered to be non-irritating to skin.

Eye irritation

The eye irritation potential of the test article was investigated by instillation of 0.1 g into one eye of each of three young adult rabbits (according to OECD guideline 405). The treated eyes were not rinsed after application. Scoring of irritation effects were performed 1, 24, 48 and 72 hours after test article application. Administration of the test article into the conjunctival sac of the rabbit eye induced slight erythema and swelling. These reactions were fully reversible after 24h. Therefore, the test substance is considered to be non-irritating to eyes.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. A GLP-compliant OECD 404 study is available for skin irritation. A GLP-compliant OECD 405 study is available for eye irritation. In both studies, the scores for the test item treated tissues were below the thresholds for classification as an irritant. As a result the substance is not considered to be classified for skin or eye irritation under Regulation (EC) No. 1272/2008,as amended for the thirteenth time in Regulation (EC) No 2018/1480.