Dibutyl fumarate

Endpoint summary

Description of key information

Skin irritation/corrosion in vivo: OECD 404, EU B.4: not irritating.
Eye irritation/corrosion in vivo: OECD 405, EU B.5: not irritating.

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:

1 April 1999 to 14 July 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP 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

Details on test animals or test system and environmental conditions:

Species and strain: Rabbits, New Zealand White.Supplier: J. Schitkovits, A-2620 Natschbach.Number and sex: 3 females.Body weight (at the start and at the termination of the study):Animal No. 101: 3.1 kg and 3.4 kg.Animal No. 102: 2.8 kg and 3.1 kg.Animal No. 103: 2.4 kg and 2.7 kg.Hygiene: Optimal hygienic conditions.Room number: EH1-17.Room temperature: Average of 20 °C.Relative humidity: Average of 55 %.Air exchange: 12 per hour.Light: Artificial light from 6 a.m. to 6 p.m.Cages: Individual caging in metal wire cages, Ehret GmbH, D-79312 Emmendingen, type KK 016R, 79 cm x 59 cm x 35 cm.Feed: Altromin 2023 diet for rabbits, ad libitum. Random samples of the feed are analysed for contaminants by Altromin, D-32791 Lage.Water: Tap water from an automatic watering system, ad libitum.Identification: Labelling with felt-tipped pen in a pinna. Acclimatisation: 11 days.

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 mL per animal

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48, 72 h and 6 and 8 d after patch removal (ending individually).

Number of animals:

3

Details on study design:

Hair was clipped on the dorsal area of the trunk one day before the application. An electric hair clipper, Aesculap GH 204, with a 1 mm cutterhead was used.The test sites were located on the dorsal thoracal region.Samples of 0.5 ml of the test substance were spread on cellulose patches 1) in a size of about 2.5 cm x 2.5 cm and were applied to the test sites. They were held in place by fixing them marginally with non irritating tapes 2). The application sites were covered semi-occlusively by a dressing 3).Access by the animal to the application sites was prevented by a plastic collar.The duration of exposure was 4 hours.At the end of the exposure period the dressing, the tapes with the patches and the collar were removed, residual test substance was removed with wet Kleenex-tissues, if necessary.1) Pur Zellin-Tupfer, obtained by Fa. Hartmann , A-2355 Wiener Neudorf.2) "Blenderm ®" surgical tape, hypoallergenic, 3M, Medical Products Division, St. Paul, MN 551444, USA.3) Self adhesive non woven fabric, hypoallergenic, made by Beiersdorf AG, D-20245 Hamburg.The treated areas and the surrounding untreated skin (control area) of the animals were examined for erythema/eschar and oedema as well as for other local alterations approximately 1, 24, 48, 72 h and 6 and 8 d after patch removal (ending individually).The scores noted, 24, 48 and 72 h after the patch removal were decisive for the interpretation of the results and the conclusion.The skin was examined using a cold light source KL 1500 electronic.Dermal irritation was described and recorded according to the scoring scheme for acute dermal irritation/corrosion tests.

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h, 6d

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 6 d

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h, 6d, 8d

Score:

1.3

Max. score:

2

Reversibility:

fully reversible within: 8 d

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h, 6d, 8d

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 8 d

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h, 6d

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 6 d

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h, 6d, 8d

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h, 6d, 8d

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 6 d

Irritant / corrosive response data:

All areas treated with the test substance and all control areas were normal before the application.The control areas were normal at any observation time.Erythema/eschar: Very slight erythema to well-defined erythema (scores "1" and "2") were noted in all animals. The lesions were observed from 1 h p.a. to a maximum of 6 d p.a.Oedema: Very slight oedema (score "1") and moderate oedema (score "3") was noted in 1/3 animals each. The lesions then gradually returned to normal and 6 d p.a. no swellings were detectable.

Other effects:

None

 

	Mean Scores for Animal No.:
	101	102	103
Erythema / Eschar:	2.0	1.3	2.0
Oedema:	3.0	0.0	1.0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test substance caused transient skin irritation. Based on the mean scores calculated from the individual examinations performed 24, 48 and 72 h p.a. and referring to Commission Directive 2001/59/EC the test substance does require classification for skin irritation (R 38 - Irritating to skin).

Executive summary:

Aim of the study

The aim of this study was to investigate possible irritation or corrosion by the test material following a single application to the intact skin of rabbits.

Methods

Methods and investigations were performed in conformance with the OECD Guideline 404 and the Directive 92/69/EEC, method B.4.

Administration of the test substance

0.5 ml "DIBUTYLFUMARAT" was applied via a patch to a site of about 2.5 cm x 2.5 cm of the intact skin of each of 3 rabbits and covered by semi-occlusive dressing.

The duration of the exposure was 4 hours.

Investigations

·     Body weights: at the start and at the termination of the test.

·     General signs of toxicity: once daily.

·     Skin examination: 1, 24, 48, 72 h and 6 and 8 d after patch removal (ending individually).

Results

General signs of toxicity

No general toxic effects were observed.

Skin examination

Very slight to well-defined erythema was noted in all animals. The lesions lasted until a maximum of 6 days after the patch removal.

Moderate oedema was seen in one animal and very slight oedema in another animal, lasting until a maximum of 72 hours after the end of the exposure.

Conclusion

The following mean scores were calculated for each animal from the individual examinations performed 24 h, 48 h and 72 h p.a.:

 

	Mean Scores for Animal No.:
	101	102	103
Erythema / Eschar:	2.0	1.3	2.0
Oedema:	3.0	0.0	1.0

 

Lesions were reversible in all animals within 8 days p.a.

  

The test substance

"DIBUTYLFUMARAT"

caused transient skin irritation. Based on the mean scores calculated from the individual examinations performed 24, 48 and 72 h p.a. and referring to Commission Directive 2001/59/EC the test substance does require classification for skin irritation (R 38 - Irritating to skin). No classification is derived under CLP, as none of the criteria for classification in category 2 is met.

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:

1 April 1999 to 14 July 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP 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

Details on test animals or tissues and environmental conditions:

Species and strain: Rabbits, New Zealand WhiteSupplier: J. Schitkovits, A-2620 NatschbachNumber and sex: 3 femalesBody weight (at the start and at the termination of the test):Animal No. 111: 2.7 kg and 2.8 kg.Animal No. 112: 2.8 kg and 2.9 kg.Animal No. 113: 3.4 kg and 3.5 kg.Hygiene : Optimal hygienic conditions.Room number: EH1-17.Room temperature: Average of 20 °C.Relative humidity: Average of 57 %.Air exchange: 12 per hour.Light: Artificial light from 6 a.m. to 6 p.m.Cages: Individual caging in metal wire cages, Ehret GmbH, D-79312 Emmendingen, type KK 016R, 79 cm x 59 cm x 35 cm.Feed: Altromin 2023 diet for rabbits, ad libitum. Random samples of the feed are analysed for contaminants by Altromin, D-32791 Lage.Water: Tap water from an automatic watering system, ad libitum.Identification: Labelling with a felt-tipped pen in an auricle. Acclimatisation: 10 or 16 days.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL per animal

Observation period (in vivo):

1, 24, 48 and 72 hours

Number of animals or in vitro replicates:

3

Details on study design:

0.1 ml of the test substance was administered into the conjunctival sac of the right eye by gently pulling the lower lid away from the eyeball for instillation. The eye was held closed for about one second to prevent a loss of test substance.The left eye remained untreated and served as a control.Firstly, the test substance was given to one animal. As there was no evidence for a serious damage to the eye of this animal noted until 72 h p.a., the test substance was administered to the other two animals too.Both eyes of the animals were examined within 24 hours before the instillation and approximately 1, 24, 48 and 72 hours p.a.No examinations were performed thereafter.The scoring terms of 24, 48 and 72 h after the instillation were decisive for the interpretation of the results and the conclusion of the study.The whole eye, especially the cornea, the iris and the conjunctivae were examined, using an otoscope lamp. Eye irritation was scored and recorded using the scheme for eye lesions.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

all animals

Time point:

other: 1, 24, 48, 27 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

mean

Remarks:

all animals

Time point:

other: 1, 24, 48, 72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 24 h

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 24 h

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

mean

Time point:

other: 1, 24, 48, 72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Irritant / corrosive response data:

All eyes, treated with the test substance ("test eyes") were normal before the instillation.All untreated eyes ("control eyes") were normal at any observation time.Test eyes:Corneae and Irises: No lesions were seen in any animal at all examination terms.Conjunctivae, redness: All animals were affected 1 h or 24 h or 48 p.a., with a score of "1". No changes were observed later on.Conjunctivae, chemosis: 1/3 animals was affected 1 h p.a., with a score of "1". No lesions were seen at the other animals and at the other examination terms.

Other effects:

None

   

	Mean Scores for Animal No.:
	111	112	113
Cornea:	0.0	0.0	0.0
Iris:	0.0	0.0	0.0
Conjunctivae, redness:	0.3	0.3	0.0
Conjunctivae, chemosis:	0.0	0.0	0.0

 Scores of the test eyes.

Examination of cornea and iris. Individual data and means.

Time after instillation (p.a.)	Cornea animal No.			Iris animal No.
	111	112	113	111	112	113
1 h	0	0	0	0	0	0
24 h	0	0	0	0	0	0
48 h	0	0	0	0	0	0
72 h	0	0	0	0	0	0
mean (24-72 h)	0.0	0.0	0.0	0.0	0.0	0.0

 

Examination of conjunctivae. Individual data and means.

Time after instillation (p.a.)	Conjunctivae                     Redness                                           Chemosis                    animal No.                                          animal No.
	111	112	113	111	112	113
1 h	0	0	1	0	1	0
24 h	1	0	0	0	0	0
48 h	0	1	0	0	0	0
72 h	0	0	0	0	0	0
mean (24-72 h)	0.3	0.3	0.0	0.0	0.0	0.0

Interpretation of results:

not irritating

Remarks:

Migrated informationCriteria used for interpretation of results: EU

Conclusions:

The test substance caused slight transient ocular irritation. Based on the mean scores calculated from the individual examinations performed 24, 48 and 72 h p.a. and referring to Commission Directive 2001/59/EC the test substance does not require classification for eye irritation.

Executive summary:

Aim of the study

The aim of this study was to investigate possible irritating or corrosive effects of the test substance following a single administration into the conjunctival sac of one eye of rabbits.

Methods

Methods and investigations were performed in conformance with the OECD-Guideline 405 and the Directive 92/69/EEC, method B.5.

Administration of the test substance

0.1 ml of "DIBUTYLFUMARAT" was instilled into the conjunctival sac of one eye of each of 3 rabbits.

Investigations

·     Body weights: at the start and at the termination of the test.

·     General signs of toxicity: once daily.

·     Eye examination: 1, 24, 48 and 72 h p.a.

Results

General signs

No other than ocular alterations were noted.

Eye examination

·     Corneae and irises were not affected.

·  Conjunctivae: Some injected blood vessels were observed in all animals and a slight swelling in 1/3 animals at 1, 24 or 48 hours p.a.

Conclusion

The following mean scores were calculated for each animal from the individual examinations performed 24 h, 48 h and 72 h p.a.:

 

	Mean Scores for Animal No.:
	111	112	113
Cornea:	0.0	0.0	0.0
Iris:	0.0	0.0	0.0
Conjunctivae, redness:	0.3	0.3	0.0
Conjunctivae, chemosis:	0.0	0.0	0.0

 

Ocular lesions were reversible in all animals within 72 hours p.a.

  

The test substance

DIBUTYLFUMARAT

is only slightly irritating to the eyes. Based on the mean scores calculated from the individual examinations performed 24, 48 and 72 h p.a. and referring to Commission Directive 2001/59/EC the test substance does not require classification for eye irritation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Justification for classification or non-classification

According to the criteria of the CLP, DBF was not irritating to the skin, and not irritating to the eyes of rabbits.