1,3,5-tris[[4-tert-butyl-3-hydroxy-2,6-xylyl]methyl]-1,3,5-triazine-2,4,6(1H,3H,5H)-trione

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April - June 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Also according to GLP

Justification for type of information:

Until 2016, Annex IX of REACH required an in vivo test for eye irritation to confirm negative results of in vitro tests. In the interest of animal welfare and to minimize any testing likely to produce severe responses in animals, a weight of evidence analysis was performed, prior to the start of this in vivo eye irritation study in the rabbit. As recommended in the test guidelines, all available information was evaluated (e.g. existing human and animal data, literature, substance data, analysis of structure-activity relationships, physicochemical properties and reactivity (pH, buffering capacity) and in vitro, ex-vivo and in vivo tests) to determine the need for in vivo eye testing.
Because no skin effects were noted in the in vivo skin irritation study and since a negative result was anticipated for the possible in vitro study, it was concluded that there is a need to perform this in vivo eye irritation study in rabbit in order to establish the possible eye irritating properties of the test substance.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1973

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Pre-current guideline, pre GLP study.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Animals were dosed orally with test material in a dosing solution, and observed for 14 days..

GLP compliance:

no

Test type:

other: single oral dose

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

Ten nonfasted male Harlan-Wistar albino rats (90-120g)

Route of administration:

oral: unspecified

Vehicle:

other: agar and Tween 80

Details on oral exposure:

Test animals were treated with a single oral dose of 10.0 g/kg. The dosing solution contained 0.2 g test material in a total volume of 1 ml of 0.25% agar and 0.10% Tween 80.

Doses:

Single oral dose of 10.0 g/kg

No. of animals per sex per dose:

Ten males

Control animals:

no

Details on study design:

Animals were observed for toxicity over a period of 14 days, after which they were euthanized and subjected to gross necropsy.

Statistics:

No statistics.

Sex:

male

Dose descriptor:

LD50

Effect level:

> 10 000 mg/kg bw

Based on:

test mat.

Mortality:

None

Clinical signs:

other: No adverse signs.

Gross pathology:

All organs appeared normal at necropsy.

Other findings:

There were no signs of intoxication

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of this study, an LD50 of > 10 000 mg/kg bw has been calculated for males for the test material.

Executive summary:

In an acute oral toxicity study, the test item was administered to a group of 10 male Sprague Dawley rats at a single dose level of 10 000 mg/kg bw.

No mortalities or clinical signs were noted.

In conclusion, the LD50 of the test item is determined to be > 10 000 mg/kg body weight by oral route in the rat.

Cross-reference 2

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1973

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Not a guideline study. Predated GLP. Male animals only tested. Otherwise adequate study.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Standard acute dermal study of the period.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rabbit

Strain:

other: albino

Sex:

male

Details on test animals or test system and environmental conditions:

5 male albino rabbits (avg. wt 2492 g) were used.

Type of coverage:

other: The test site was then covered (the material used was not listed). It therefore is not known whether the application was occlusive or semi-occlusive.

Vehicle:

water

Details on dermal exposure:

Test material was applied to clipped skin of 5 male albino rabbits (avg. wt 2492 g). The material that was applied was weighed onto Vinylite and moistened with a volume of water equal to two times the weight of the test material). The test site was then covered (the material used was not listed). Animals were observed for 14 days, after which they were weighed, euthanized and subjected to gross necropsy.

Duration of exposure:

14 days

Doses:

One dose level: 5000 mg/kg

No. of animals per sex per dose:

Five males

Control animals:

no

Details on study design:

Test material was applied to clipped skin of 5 male albino rabbits (avg. wt 2492 g). The material that was applied was weighed onto Vinylite and moistened with a volume of water equal to two times the weight of the test material). The test site was then covered (the material used was not listed). Animals were observed for 14 days, after which they were weighed, euthanized and subjected to gross necropsy.

Statistics:

None employed

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths.

Clinical signs:

other: There were no signs of intoxication or irritation. No clinical effects were noted.

Gross pathology:

There were no remarkable necropsy findings.

Other findings:

None noted.

Interpretation of results:

GHS criteria not met

Conclusions:

No mortalities; LD50 >5000 mg/kg.

Executive summary:

An acute dermal toxicity test with male rabbits was performed. Male rabbits were exposed to 5000 mg/kg bw and observed for 14 days after removal of the test substance. No mortality, clinical signs or particular findings at necropsy were observed.

Based on these results, the LD50 of the test substance was found to be > 5000 mg/kg bw and therefore the test substance is not classified.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,3,5-tris[[4-tert-butyl-3-hydroxy-2,6-xylyl]methyl]-1,3,5-triazine-2,4,6(1H,3H,5H)-trione

EC Number:

254-996-9

EC Name:

1,3,5-tris[[4-tert-butyl-3-hydroxy-2,6-xylyl]methyl]-1,3,5-triazine-2,4,6(1H,3H,5H)-trione

Cas Number:

40601-76-1

Molecular formula:

C42H57N3O6

IUPAC Name:

tris[(4-tert-butyl-3-hydroxy-2,6-dimethylphenyl)methyl]-1,3,5-triazinane-2,4,6-trione

Constituent 2

Reference substance name:

Cyanox (TM) 1790 Antioxidant

IUPAC Name:

Cyanox (TM) 1790 Antioxidant

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: at least 6 weeks old
- Weight at study initiation: at least 1 kg
- Housing: cages with perforated floors and shelters
- Diet: ca. 100 g per day of pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy). Hay and wooden sticks were available during the study period
- Water: free access to tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24
- Humidity (%): 40-70
- Air changes (per hr): ca. 15
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: one eye remained untreated and served as reference control

Amount / concentration applied:

TEST MATERIAL
Amount applied: 74 mg (= approx. 0.1 ml)

Duration of treatment / exposure:

single instillation

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 males

Details on study design:

STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner three weeks later, after considering the degree of eye irritation observed in the first animal.

TREATMENT
Animals were treated by instillation of, on average, 74.0 mg (range 73.9 - 74.2 mg) of the test substance (a volume of approximately 0.1 mL) in the conjunctival sac of one of the eyes 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 substance.

REMOVAL OF TEST SUBSTANCE
-Washing (if done): No

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to instillation) and after the final observation.
- Necropsy: No necropsy was performed according to protocol.
- Irritation: The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 days after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to OECD 405.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Results 2

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Reversibility:

fully reversible within: 48h

Results 3

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Reversibility:

fully reversible within: 7d

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Reversibility:

fully reversible within: 7d

Results 6

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Reversibility:

fully reversible within: 48h

Results 7

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Results 8

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Results 9

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Reversibility:

fully reversible within: 48h

Results 10

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Results 11

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Results 12

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritant / corrosive response data:

Effects on cornea, iris and conjunctivae were observed:
The corneal injury consisted of slight dulling of the normal lustre on Day 1 and no epithelial damage was noted. Iridial irritation grade 1 was observed and resolved within 24 hours in two animals and within 48 hours in one animal. The irritation of the conjunctivae consisted of redness, chemosis and discharge and completely resolved within 48 hours in one animal and within 7 days in the other two animals.

Other effects:

No staining of ocular tissues by the test substance was oberved. Remnants of the test substance were present in the eye and on the outside of the eyelids in all three animals at 1 hour after exposure.
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the above results, the test substance does not have to be classified and has no obligatory labelling requirement for eye irritation.

Executive summary:

An eye irritation study was performed according to OECD TG 405 and under GLP conditions. An instillation of 74 mg of the test item in one eye of 3 rabbits. One eye remained untreated and served as a control. No irreversible effects were noted.

Based on the results above, the test material is not classified for eye irritation.