4-oxo-4-(p-tolyl)butyric acid adduct with 4-ethylmorpholine

Endpoint summary

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:

05.05.1995 - 08.05.1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Three New Zealand White rabbits supplied by David Percival Ltd., Moston, Sandbach, Cheshire, UK were used. At the start of the study the animals weighed 2.60 to 2.65 kg and were twelve to sixteen weeks old. After a minimum acclimatisation period of five days each animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.
The animals were individually housed in suspended metal cages. Free access to mains drinking water and food (STANRAB SQC Rabbit Diet, Special Diets Services Ltd., Witham, Essex, UK) was allowed throughout the study.
The animal room was maintained at a temperature of 18 to 22 °C and relative humidity of 58 to 66%. The rate of air exchange was approximately 15 changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

no

Amount / concentration applied:

0.5 g of the test material, moistened with 0.5 mL of distilled water

Duration of treatment / exposure:

4 h

Observation period:

1h, 24 h, 48 h and 72 h

Number of animals:

3

Details on study design:

On the day before the test each rabbit was clipped free of fur from the dorsal flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.
On the day of the test a suitable test site was selected on the back of each rabbit. A quantity of 0.5 g of the test material, moistened with 0.5 mL of distilled water, was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the shorn skin. The patch was secured in position with a strip of surgical adhesive tape (BLENDERM: approximate size 2.5 cm x 4.0 cm). To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset (TUBIGRIP) and the animals were returned to their cages for the duration of the exposure period.
Four hours after application the corset and patches were removed from each animal and any residual test material removed by gentle swabbing with cotton wool soaked in distilled water.
Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the followong scale from Draize J. H., (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington D. C., p. 31.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

48 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 1 h

Score:

1

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

48 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 1 h

Score:

1

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

48 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

48 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 1 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

48 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 1 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

48 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Reversibility of any observed effect: Changes fully reversible within 1 days

Interpretation of results:

GHS criteria not met

Conclusions:

The test material produced a primary irritation index of 0.0 and was classified as NON-IRRITANT to rabbit skin according to Draize classification scheme. No corrosive effects were noted.

Executive summary:

A study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The method used followed the recommendations of the OECD Guidelines for Testing of Chemicals No. 404 "Acute Dermal Irritation/Corrosion" (adopted 17 July 1992) and Method B4 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

The results may be used as a basis for classification and labelling under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 93/21/EEC) relating to the classification, packaging and labelling of dangerous substances.

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced very slight erythema confined to two treated skin sites one hour after patch removal. No dermal reactions were noted at the 24-hour observation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation, other

Remarks:

in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17.05.1995 - 19.05.1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

evaluation of eye response refering to Draize's method: 1959

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

One New Zealand White rabbit supplied by David Percival Ltd., Moston, Sandbach, Cheshire, UK, was used. At the start of the study the animal weighed 3.08 kg and was approximately twelve to six weeks old. After a minimum acclimatisation period of five days the animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.
The animal was housed in a suspended metal cage. Free access to mains drinking water and food (STANRAB SQC Rabbit Diet, Special Diets Services Ltd., Witham, Essex, UK) was allowed throughout the study.
The animal room was maintained at a temperature of 17 to 20 °C and relative humidity of 49 to 58%. The rate of air exchange was approximately 15 changes per hour and the lighting was controlled by a time switch to give 12 hours continous light and 12 hours darkness.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

about 1 s

Observation period (in vivo):

1 h, 24 h, 48 h

Number of animals or in vitro replicates:

1 animal

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbit was examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope.
A volume of 0.1 mL of the test material, which was found to weigh approximately 17 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released. The left eye remained and was used for control purposes. Immediately after administration of the test material, an assassment of the initial pain reaction was made.
Assessment of ocular damage/irritation was made approximately 1, 24 and 48 hours following treatment, according to the numerical evaluation given in Appendix I, (from Draize J.H. (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Acedemy of Sciences, Washington D.C. p.48 to 49).
Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 1 h

Score:

5

Max. score:

10

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24 h

Score:

5

Max. score:

10

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

iris score

Basis:

animal #1

Time point:

48 h

Score:

5

Max. score:

10

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 1 h

Score:

40

Max. score:

80

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24 h

Score:

40

Max. score:

80

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

48 h

Score:

40

Max. score:

80

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 1 h

Score:

2

Max. score:

6

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24 h

Score:

2

Max. score:

6

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

48 h

Score:

2

Max. score:

6

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 1 h

Score:

2

Max. score:

8

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24 h

Score:

2

Max. score:

8

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

48 h

Score:

2

Max. score:

8

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

other: discharge score

Basis:

animal #1

Time point:

other: 1 h

Score:

3

Max. score:

6

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

other: discharge score

Basis:

animal #1

Time point:

24 h

Score:

2

Max. score:

6

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

other: discharge score

Basis:

animal #1

Time point:

48 h

Score:

3

Max. score:

6

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

maximum mean total score (MMTS)

Basis:

animal #1

Time point:

other: 1 h

Score:

59

Max. score:

110

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

maximum mean total score (MMTS)

Basis:

animal #1

Time point:

24 h

Score:

57

Max. score:

110

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritation parameter:

maximum mean total score (MMTS)

Basis:

animal #1

Time point:

48 h

Score:

59

Max. score:

110

Reversibility:

other: On humane grounds the study was determined prematurely after the 48 hour observation time due to the severity of the eye irritation.

Irritant / corrosive response data:

Areas of translucent corneal opacity were noted in the treated eye one hour after treatment and at the 24 and 48-hour observations. Sloughing of the cornea was noted in the treated eye one hour after treatment.

Other effects:

Sloughing of the cornea and lower conjunctival membrane together with pale green discolouration of the nictitating membrane was observed.

Iridial inflammation was noted in the treated eye one hour after treatment and at the 24 and 48-hour observations.
Moderate conjunctival irritation was noted in the treated eye one hour after treatment and at the 24 and 48-hour observations.
Pale green discolouration of the nicitating membrane was noted in the treated eye one hour after treatment. Sloughing of the lower conjunctival membrane was noted in the treated eye at the 48-hour observation.

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The test material produced a maximum score of 59.0 and was classified as at least a severe irritant (Class 6 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.
The test material was also considered to be irritant according to EU labelling regulations. It is reasonable to assume that the symbol "Xi", the indication of danger "irritant" and the highest risk phrase R41 "RIKS OF SERIOUS DAMAGE TO EYES" are therefore required.

Executive summary:

A study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion" (adopted 24 February 1987) and Method B5 of Comission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

The results may be used as a basis for classification and labelling under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 93/21/EEC) relating to the classification, packaging and labelling of dangerous substances.

A single application of the test material to the non-irrigated eye of one rabbit produced areas of translucent corneal opacity, iridial inflammation and moderate conjunctival irritation. Other ocular effects noted were sloughing of the cornea and lower conjunctival membrane and pale green discolouration of the nicitating membrane. The animal was killed for humane reasons immediately after the 48-hour observation in accordance with current UK Home Office guidelines. No further animals were treated.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)