5,7-Di-t-butyl-3-[3,5-dimethyl-4-[(1,3,7,9-tetra-t-butyl-5-methyl-5H-benzo[d][1,3,2]benzodioxaphosphocin-11-yl)oxy]phenyl]-3H-benzofuran-2-one

Endpoint summary

Description of key information

Based on results of a GLP and OECD 404 compliant in vivo study, the test item is not considered to be skin irritating in rabbits. Furthermore, based on results of a GLP and OECD 405 compliant in vivo study, the test item is not considered to be eye irritating in rabbits.

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:

2016-06-08 to 2016-09-20

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:

2015

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nousan No. 8147, as this is in line with OECD 404

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature; under light exclusion ; no direct sunlight

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo CRS (Switzerland) Limited
- Age at study initiation: approx. 7-9 months
- Weight at study initiation: 3.70 - 4.51 kg
- Housing: individual housing
- Diet: STANRAB (P) SQC; SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 +/- 3 °C
- Humidity: 30 - 70 %
- Air changes: approx. 10 per hr
- Photoperiod: 12 /12 hrs dark / hrs light

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g was loaded on the test patch (2.5 x 2.5 cm)

Duration of treatment / exposure:

4 h

Observation period:

72 h

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: dorsolateral part of the trunk
- Type of wrap if used: The item was covered with a test patch and adhesive fleece

REMOVAL OF TEST SUBSTANCE
- Washing: The test item was removed at the end of the exposure period with polyethylenglycol
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
Immediately after removal of the patch, approx. 1, 24, 48, 72 h after removal of the patch

SCORING SYSTEM: Evaluation of skin reactions was performed according to the quoted guidelines.
- Method of calculation: For evaluation mean values of erythema and edema were calculated based on readings at 24, 48, 72 h. Calculations were performed using Microsoft Excel 2010 and checked with a calculator.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Very slight erythema (grade 1) was observed in two animals immediately after removal of the patch and persisted until hour 1 or hour 24. The third animal was free of any findings. The cutaneous reactions were reversible in two animals within 24 or 48 hours after removal of the patch.

Other effects:

- Other adverse local effects: none
- Other adverse systemic effects: none

Table 1. Tabulation of irritant response data for each individual animal at each observation time as well as calculation of the means

Readings	Animal No.	Erythema	Edema
0 h	1	1	0
	2	0	0
	3	1	0
1 h	1	1	0
	2	0	0
	3	1	0
24 h	1	1	0
	2	0	0
	3	0	0
48 h	1	0	0
	2	0	0
	3	0	0
72 h*	1	0	0
	2	0	0
	3	0	0
Mean 24 h – 72 h	1	0.3	0.0
	2	0.0	0.0
	3	0.0	0.0
Mean	-	0.1	0.0

*Study discontinued because the animals were free of findings

Interpretation of results:

GHS criteria not met

Conclusions:

Considering the described cutaneous reactions as well as the average score for irritation, the test item does not show a skin irritating potential under the test conditions chosen.

Executive summary:

The potential of the test item to cause acute dermal irritation or corrosivity was assessed by a single topical application of an amount of 0.5 g of the test item for 4 hours to the intact skin of three New Zealand White rabbits (stepwise procedure starting with one animal and supplementing two additional animals), using a patch of 2.5 cm x 2.5 cm, covered with semi-occlusive dressing. After removal of the patch the application area was washed off. The cutaneous reactions were assessed immediately after removal of the patch, approximately 1, 24, 48 and 72 hours after removal of the patch. The following test item-related clinical observations were recorded during the course of the study: Very slight erythema (grade 1). The cutaneous reactions were reversible in two animals within 24 or 48 hours after removal of the patch. The third animal did not show any findings. Mean scores over 24, 48 and 72 hours for each animal were 0.3, 0.0 and 0.0 for erythema and 0.0, 0.0 and 0.0 for edema. Considering the described cutaneous reactions as well as the average score for irritation, the test article does not show a skin irritating potential under the test conditions chosen.

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:

2016-06-08 to 2016-09-20

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:

2012

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan/MAFF 8147

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature; under light exclusion ; no direct sunlight

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Envigo CRS (Switzerland) Limited
- Age at study initiation: approx. 3 months
- Weight at study initiation: 2.49 - 2.81 kg
- Housing: individual housing
- Diet: STANRAB (P) SQC; SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 +/- 3 °C
- Humidity: 30 - 70 %
- Air changes: approx. 10 per hr
- Photoperiod: 12 / 12 hrs dark / hrs light

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL (about 17 mg of the comminuted test item)

Duration of treatment / exposure:

24 h

Observation period (in vivo):

approx. 1, 24, 48, 72 and 96 h after application

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: rinsing with 3 to 6 mL of lukewarm tap water for 1 to 2 minutes using a syringe with a blunt probe
- Time after start of exposure: 24 h

SCORING SYSTEM: according to guidelines and, in addition, evaluations were made of discharge from the eye and the area of cornea affected by the lesions:
Area of cornea involved (ar) Grading:
1 > 0 ≤ ¼
2 > ¼ < ½
3 > ½ < ¾
4 > ¾

Discharge (di) Grading:
0 No discharge
1 Slight: any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2 Obvious: discharge with moistening of the lids and hairs just adjacent to lids
3 Severe: discharge with moistening of the lids and hairs, and considerable area around the eye


TOOL USED TO ASSESS SCORE: hand-slit lamp, otoscope lamp and fluorescein

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

No corneal lesions were noted in any animal at any examination time point. Even after instillation of a fluorescein solution no corneal lesions were detectable after 24 and 48 hours. Moderate iritis (grade 1) was observed in one animal 1 hour after application only. Slight conjunctival redness (grade1) was noted in all animals from 1 hour until hour 24 after application. Slight conjunctival chemosis (grade 1) was noted in two animals 1 hour after application. Additional findings like injected scleral vessels in a circumscribed or circular area were noted within 48 hours after application. The ocular reactions were reversible in all animals within 48 hours after application.

Other effects:

- Lesions and clinical observations: none
- Ophthalmoscopic findings: none
- Histopathological findings: not performed
- Effects of rinsing or washing: none

Table 1. Irritant response, data for each individual animal at each observation time as well as calculation of the mean

		Cornea		Iris	Conjunctiva
Readings	Animal No.	Opacity	Area of cornea involved		Redness	Chemosis	Discharge
1 h	1	0	0	0	1	0	0
	2	0	0	1	1	1	0
	3	0	0	0	1	1	0
24 h	1	0	0	0	1	0	0
	2	0	0	0	1	0	0
	3	0	0	0	1	0	0
48 h	1	0	0	0	0	0	0
	2	0	0	0	0	0	0
	3	0	0	0	0	0	0
72 h	1	0	0	0	0	0	0
	2	0	0	0	0	0	0
	3	0	0	0	0	0	0
96 h	1	0	0	0	0	0	0
	2	0	0	0	0	0	0
	3	0	0	0	0	0	0
Mean 24 h- 72h	1	0.0	-	0.0	0.3	0.0	-
	2	0.0	-	0.0	0.3	0.0	-
	3	0.0	-	0.0	0.3	0.0	-
Mean	-	0.0	-	0.0	0.3	0.0	-

Interpretation of results:

GHS criteria not met

Conclusions:

Considering the described ocular reactions as well as the average score for irritation, the test item does not show an eye irritating potential under the test conditions chosen.

Executive summary:

The potential of the test item to cause damage to the conjunctiva, iris or cornea was assessed by a single ocular application of 0.1 mL bulk volume (about 17 mg) of the undiluted test item to one eye of three New Zealand White rabbits (stepwise procedure starting with one animal and supplementing two additional animals). About, and not less than 24 hours after application the eye was rinsed with tap water. The ocular reactions were assessed approximately 1, 24, 48, 72 and 96 hours after application. Additional eye examinations were performed at 24 and 48 h after application with the instillation of a fluorescein solution. Due to absence of detectable lesions following fluorescein instillation, all subsequent readings were performed without the aid of fluorescein. No corneal lesions were noted in any animal at any examination time point. Even after instillation of a fluorescein solution no corneal lesions were detectable after 24 and 48 hours. The following test item-related clinical observations were recorded during the course of the study: Moderate iritis (grade 1), slight conjunctival redness (grade 1) and slight conjunctival chemosis (grade 1). Additional findings like injected scleral vessels in a circumscribed or circular area were noted in the animals within the first 48 hours. The ocular reactions were reversible in all animals within 48 hours after application. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0, 0.0 and 0.0 for corneal opacity and for iris lesions, 0.3, 0.3 and 0.3 for redness of the conjunctiva and 0.0, 0.0 and 0.0 for chemosis. Considering the described ocular reactions as well as the average score for irritation, the test article does not show an eye irritating potential under the test conditions chosen.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation in vivo

The potential of the test item to cause acute dermal irritation or corrosivity was assessed by a single topical application of an amount of 0.5 g of the test item for 4 hours to the intact skin of three New Zealand White rabbits (stepwise procedure starting with one animal and supplementing two additional animals), using a patch of 2.5 cm x 2.5 cm, covered with semi-occlusive dressing. After removal of the patch the application area was washed off. The cutaneous reactions were assessed immediately after removal of the patch, approximately 1, 24, 48 and 72 hours after removal of the patch. Very slight erythema (grade 1) was observed in two animals. The cutaneous reactions were reversible in both animals within 24 or 48 hours after removal of the patch.The third animal did not show any findings. Mean scores over 24, 48 and 72 hours for each animal were 0.3, 0.0 and 0.0 for erythema and 0.0, 0.0 and 0.0 for edema. Considering the described cutaneous reactions as well as the average score for irritation, the test item does not show a skin irritating potential under the test conditions chosen.

Eye irritation in vivo

The potential of the test item to cause damage to the conjunctiva, iris or cornea was assessed by a single ocular application of 0.1 mL bulk volume (about 17 mg) of the undiluted test item to one eye of three New Zealand White rabbits (stepwise procedure starting with one animal and supplementing two additional animals). About, and not less than 24 hours after application the eye was rinsed with tap water. The ocular reactions were assessed approximately 1, 24, 48, 72 and 96 hours after application. Additional eye examinations were performed at 24 and 48 h after application with the instillation of a fluorescein solution.

Due to absence of detectable lesions following fluorescein instillation, all subsequent readings were performed without the aid of fluorescein.

No corneal lesions were noted in any animal at any examination time point. Even after instillation of a fluorescein solution no corneal lesions were detectable after 24 and 48 hours. The following test item-related clinical observations were recorded during the course of the study:

- Moderate iritis (grade 1)

- Slight conjunctival redness (grade 1)

- Slight conjunctival chemosis (grade 1)

 Additional findings like injected scleral vessels in a circumscribed or circular area were noted in the animals within the first 48 hours.

The ocular reactions were reversible in all animals within 48 hours after application. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0, 0.0 and 0.0 for corneal opacity and for iris lesions, 0.3, 0.3 and 0.3 for redness of the conjunctiva and 0.0, 0.0 and 0.0 for chemosis.

Considering the described ocular reactions as well as the average score for irritation, the test item does not show an eye irritating potential under the test conditions chosen.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. The test item revealed no eye irritating nor skin irritating potential in two in vivo studies, respectively. As a result the substance is not classified as skin or eye irritant under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.