A mixture of: esters of C14-C15 branched alcohols with 3,5-di-t-butyl-4-hydroxyphenyl propionic acid; C15 branched and linear alkyl 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoate; C13 branched and linear alkyl 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoate

Endpoint summary

Description of key information

Skin Irritation - in vivo

NON IRRITANT for the skin.

Eye irritation - in vivo

NON IRRITANT for the eye.

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:

January 22, 1991 to January 25, 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

European Economic Community Guidelines – VI Amendment, Annex V, Directive 84/449/EEC

Deviations:

not specified

GLP compliance:

yes

Specific details on test material used for the study:

No further details specified in the study report.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species and strain: New Zealand White Rabbit
Justification for the selection of the test system: New Zealand White rabbit was chosen as lagomorph species since it is widely accepted and recommended by health Authorities as an experimental model for dermal irritation studies.
Number and sex: 3 males
Bodyweight on receipt: 2.5 – 3 kg
Age on receipt: 2-3 months
Supplier: Conelli Luciano, Via Milano, 61, 28041 Arona (Novara), Shipping slip No. 309 dated December 19, 1990
Acclimation: about 5 weeks. Animal were observed daily to ascertain their fitness for the study.
Housing:
Individual caging in air conditioned rooms.
-Temperature: 20 °C ± 2
-Air changes: about 15/hour
-Relative humidity: 50% ± 15
-Light: artificial lighting with a circadian cycle of 12 hours of light (7am – 7 pm)
-Each rabbit was caged in metal cages measuring cm, 62x47.5x38h, with stainless feeder. The cages were hung on metal racks over stainless waste system. The waste was periodically flushed away by a time programmed siphon system from above.
Animal identification: by an individual number borne on an ear stud.
Animal number: 1M, 2M and 3M
Cage identification: by cage card giving indelible experiment number, animal number and sex.

Diet: 2RB15 pelleted diet produced by Charles River Italia’s feed licensee Mucedola S.r.l., Settimo Milanese.
The declared contents, on the label, on dry matter basis (moisture 12%), were:
Crude protein 13.20%
Crude fat 2.50%
Crude fiber 22.00%
Ash 9.50%
The diet was supplemented by the Producer with vitamins and trace elements.
The diet is analysed by the Producer for nutrients and contaminants.
The diet was available “ad libitum” to the animals.

Water: from the municipal water main system.
Water is filtered and distributed “ad libitum” to the animals by an automatic value system.
Periodically drinking water is analysed for microbiologic count, heavy metals, other contaminants (e.g. solvents, pesticides) and other chemical and physical characteristics. The acceptable limits of quality of the drinking water were those defined in EEC Directive 80/778.

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 ml/animal

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 animals

Details on study design:

Administration route: epidermal
Reason for selection of administration route: possible accidental exposure for humans
Administration frequency: single
Dose administration: 0.5 ml/animal
Preparation of animals and administration of the test article:
Approximately 24 hours before the test, fur was clipped from the trunk of the animals. The clipped area was approximately 10% of the whole body surface. Care was taken not to abrade the skin.
Only animal with intact skin were used.
The test article was applied to a small area (approximately 6 sq. cm) of skin on the right side of the back of each animal. The left side, remain untreated, served as control. The treated and the control area were covered with a gauze patch which was loosely held in contact with the skin by an impermeable, non-irritant, hypoallergenic tape (Blenderm-3M Italia, Milano).
Since the test article was a solid, it was applied directly onto the gauze patch.
At the end of the exposure period the residual test substance was wiped off.
Exposure time: 4 hours
Observation of the test site: at 1, 24, 48 and 72 hours after patch removal.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

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

Reversibility:

fully reversible within: 24h

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

Reversibility:

fully reversible within: 24h

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

Reversibility:

fully reversible within: 24h

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

Reversibility:

fully reversible within: 24h

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

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Skin evaluation: A very slight erythema (graded 1) was observed in all treated animals only 1 hour after patch removal.
No dermal reactions were noted at the subsequent readings (24, 48 and 72 hours after patch removal) in any animal.

Other effects:

-Mortality: No animals died.
-Observation of clinical signs and behaviour: No clinical signs or behavioural alterations were noted in an animal.

Dermal reactions (individual)

	Observation made at:	No. of animals
		1M	2M	3M
1) Erythema and eschar	60 mins 24 hrs 48 hrs 72 hrs	1 0 0 0	1 0 0 0	1 0 0 0
2) Edema	60 mins 24 hrs 48 hrs 72 hrs	0 0 0 0	0 0 0 0	0 0 0 0

 

Interpretation of results:

GHS criteria not met

Conclusions:

The test article [ANOX BF] when administered be dermal route to rabbits under the experimental conditions adopted, is to be considered “NON IRRITANT” for the skin.

Executive summary:

Experimental data obtained from an acute dermal irritation study after a single administration of 0.5 ml of the test article [ANOX BF] in the New Zealand White rabbit (3 animals) are given in this report.

 

No deaths occurred as a result of treatment, nor were observed untoward clinical signs or behavioural alterations.

At the application site, a very slight erythema (graded 1) was observed on all treated animals only 1 hour after patch removal.

 

The test article [ANOX BF] when administered be dermal route to rabbits under the experimental conditions adopted, is to be considered “NON IRRITANT” for the 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:

January 29, 1991 to February 1, 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

European Economic Community Guidelines – VI Amendment, Annex V, Directive 84/449/EEC

Deviations:

not specified

GLP compliance:

yes

Specific details on test material used for the study:

No further details specified in the study report.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species and strain: New Zealand White rabbit
Justification for the selection of the test system: New Zealand White rabbit was chosen as lagomorph species since it is widely accepted and recommended by health Authorities as an experimental model for eye irritation studies.
Number and sex: 3 males
Bodyweight on receipt: about 2.0-3.0 kg
Age on receipt: about 2-3 months
Sullpier: Conelli Luciano, Via Milano, 61, 28041, Arnoa (Novara). Shipping slip No. 22 dated January 14, 1991
Acclimation: about 5 weeks. Animals were observed daily to ascertain their fitness for the study.
Housing: individual caging in air conditioned rooms.
-Temperature: 20 °C ± 2
-Relative humidity: 50% ± 15
-Air changes: about 15/hour
-Light: artificial lighting with a circadian cycle of 12 hours of light (7am – 7pm)
-Each rabbit was caged in metal cages measuring cm, 62x47, 5x38h, with stainless feeder. The cages were hung on metal racks over a stainless waste system. The waste was periodically flushed away by a time programmed siphon system from above.
Animal identification: by an individual number borne on an ear stud.
Animal number: 4M, 5M and 6M
Cage identification: by cage number giving indelible experiment number, animal number and sex

Diet: 2RB15 pelleted diet produced by Charles River Italia’s feed licensee Mucedola S.r.l., Settimo Mialanese.
The declared contents, on the label, on dry matter basis (moisture 12%), were:
Crude protein 13.20%
Crude fat 2.50%
Crude fiber 22.00%
Ash 9.50%
The diet was supplemented by the Producer with vitamins and trace elements.
The diet is analysed by the Producer for nutrients and contaminants.
The diet was available “ad libitum” to the animals.

Water: from the municipal water main system.
Water is filtered and distributed “ad libitum” to the animal by an automatic valve system.
Periodically drinking water is analysed for microbiologic count, heavy metals, other contaminants (e.g. solvents, pesticides) and other chemical and physical characteristics.
The acceptable limits of quality of the drinking water were those defined in EEC Directive 80/778.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 ml/animal

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 animals

Details on study design:

Administration route: ocular
Reason for selection of administration route: possible accidental exposure route for humans
Administration frequency: single
Dose administered: 0.1 ml/animal
Selection of animals and administration: both eyes of each experimental animal selected for the testing were examined within 24 hours before testing started.
Only animals without eye irritation, ocular defects or pre-existing corneal injury were used.
The test article was placed in the conjunctival sac of the right eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then in order to prevent loss of the material.
The other eye (the left), remaining untreated, served as control.
The eyes of the test animals were washed out at 24 hours following the test article instillation.
Observation of the eyes: at 1, 24, 48 and 72 hours after the test article application/
After the 24-hour reading the cornea were examined after instillation of one drop of 1% solution fluorescein and subsequent washing out with sterile saline.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24h

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

CLINICAL OBSERVATIONS
- Observation of clinical signs and behevior
No clinical signs or behavioral alterations were noted.

Other effects:

Cornea: At the direct examination of the cornea no signs were noted at any time.
At the examination after instillation of one drops of 1% sodium fluorescein, performed 24 hours after the test article application, no signs suggestive of epithelial defects were noted in any animal.
Iris: No signs were noted.
Conjunctivae: At the examination performed at 1 hour after treatment in all treated rabbits, slight redness of the conjunctivae (graded 1) was noted. At the subsequent observations carried out 24, 48 and 72 hours after treatment in all rabbits no changes were evident in this tissue.

 Eye reactions (individual)

	Observation made at	No. of animals
		4M	5M	6M
1) CORNEA	60 mins. 24 hrs 48 hrs 72 hrs	0 0 0 0	0 0 0 0	0 0 0 0
2) IRIS	60 mins. 24 hrs 48 hrs 72 hrs	0 0 0 0	0 0 0 0	0 0 0 0
3) CONJUNCTIVAE Redness	60 mins. 24 hrs 48 hrs 72 hrs	1 0 0 0	1 0 0 0	1 0 0 0
4) CONJUNCTIVAE Chemosis	60 mins. 24 hrs 48 hrs 72 hrs	0 0 0 0	0 0 0 0	0 0 0 0

 

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, the test substance when administered by ocular route to rabbits under the experimental conditions adopted, is to be considered “NON IRRITANT” for the eye.

Executive summary:

Experimental data obtained from an acute eye irritation study after a single administration of 0.1 ml of the test article [ANOX-BF] in the New Zealand White rabbit (3 animals) are given in this report.

 

No clinical signs of behavioral alterations were observed.

 

Locally the test compound induced slight conjunctival redness (graded 1) in all treated animals at the 1^st observation (1 hour). At the subsequent observations carried out 24, 48 and 72 hours after test article application no changes were evident in all rabbits.

 

At the examination after instillation of one drop of 0.1% sodium fluorescein performed 24 hours after the test article application, no signs suggestive of epithelial defects were noted in an animal.

 

In conclusion, the test substance [ANOX-BF] when administered by ocular route to rabbits under the experimental conditions adopted, is to be considered “NON IRRITANT” for the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin Irritation - in vivo

Experimental data obtained from an acute dermal irritation study after a single administration of 0.5 ml of the test article [ANOX BF] in the New Zealand White rabbit (3 animals) are given in this report.

 

No deaths occurred as a result of treatment, nor were observed untoward clinical signs or behavioural alterations.

At the application site, a very slight erythema (graded 1) was observed on all treated animals only 1 hour after patch removal.

 

The test article [ANOX BF] when administered be dermal route to rabbits under the experimental conditions adopted, is to be considered “NON IRRITANT” for the skin.

Eye irritation - in vivo

Experimental data obtained from an acute eye irritation study after a single administration of 0.1 ml of the test article [ANOX-BF] in the New Zealand White rabbit (3 animals) are given in this report.

 

No clinical signs of behavioral alterations were observed.

Locally the test compound induced slight conjunctival redness (graded 1) in all treated animals at the 1^st observation (1 hour). At the subsequent observations carried out 24, 48 and 72 hours after test article application no changes were evident in all rabbits.

At the examination after instillation of one drop of 0.1% sodium fluorescein performed 24 hours after the test article application, no signs suggestive of epithelial defects were noted in an animal.

 

In conclusion, the test substance [ANOX-BF] when administered by ocular route to rabbits under the experimental conditions adopted, is to be considered “NON IRRITANT” for the eye.

Justification for classification or non-classification

Based on the available irritating studies reported for the substance, classification for skin corrosion/irritation and eye damage/irritation in accordance with Regulation 1272/2008 is not required.