2-Butenedioic acid (Z)-, ester with 1,2-propanediol, compd. with 2-(dibutylamino)ethanol

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented and reported study fully adequate for assessment. The study was conducted according to internationally accepted technical guidelines and in compliance with GLP in a recognized contract research organization.

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:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, D-97633 Sulzfeld.
- Age at study initiation: not stated
- Weight at study initiation: 2,0-2,3 kg
- Housing: individual caging in metal wire cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 -12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): average of 19
- Humidity (%): average of 48.8
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0,5mL
- Concentration (if solution): neat test material

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 animals

Details on study design:

TEST SITE
- Area of exposure: medianon the dorsal thoracal region, 2.5 cm x 2.5 cm
- Type of wrap if used: Pur Zellin-Tupfer

SCORING SYSTEM:
Erythema I Eschar formation
0 No erythema.
1 Very slight erythema (barely perceptible).
2 Well-defined erythema.
3 Moderate to severe erythema.
4 Severe erythema (beet redness) or eschar formation (injuries in depth).

Oedema formation
0 No oedema.
1 Very slight oedema (barely perceptible).
2 Slight oedema (edges of area weil defined by definite raising).
3 Moderate oedema (raised approximately 1 mm).
4 Severe oedema (raised more than 1 mm and extending beyend area of exposure).

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

erythema score

Basis:

animal: 2 and 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

edema score

Basis:

animal: 2 and 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

No symptoms of systemic toxicity were observed in the animals during the test period and no mortalitiy occured.

All areas to be treated with the test substance and all control areas were normal before the application.The control areas were normal at any observation time.

Interpretation of results:

other: not irritating according to Regulation 1272/2008

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to internationally accepted technical guidelines and in compliance with GLP in a recognized contract research organization. The study is scientifically valid and the report is fully adequate for assessment, despite some apparent typing errors regarding the grading of ocular findings.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

of 2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

of 2004

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS

- Number and Sex: 3 males
- Animal supplier: Charles River Deutschland GmbH, D-97633 Sulzfeld, Germany
- Weight at study start or test start*: Minimum 1.9 kg, maximum 2.4 kg
- Hygiene: Optimal hygienic conditions
- Housing: Individual housing in metal wire cages from Ehret GmbH, D-79312 Emmendingen,
cage type KK 016R, 79 cm x 59 cm bottom area, 38 cm height
- Environmental enrichment: Aspen wood blocks
- Diet (ad libitum): Altromin 2123 maintenance diet for rabbits, rich in crude fiber, Altromin, D-32791 Lage, Germany
In addition, dietary supplement of hay-briquettes from SSNIFF, D-59494 Soest, Germany
- Water(ad libitum): Tap water from an automatic watering system
- Acclimation period: 5 days (animal #1), 12 days (animals #2 & #3).

Analysis of the diet for ingredients and contaminants has been performed randomly by the supplier (Altromin, D-32791 Lage, Germany)

* The study report does not clearly specify whether "study or test start" refers to prior to dosing on the day of instillation.

ENVIRONMENTAL CONDITIONS

Air conditioned animal room set at:
- Temperature (°C): Room temperature, average of 19.2°C (continuous control and recording)
- Relative Humidity (%): Average of 48.4 % (continuous control and recording)
- Photoperiod (artificial lighting): 12 hrs day / 12 hrs night

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL of undiluted liquid test material was administered into the conjunctival sac of one eye per rabbit (right eye). The contralateral eye (left eye) remained untreated to serve as a control.

Duration of treatment / exposure:

Rinsing of the eyes after treatment has not been mentioned in the report. Therefore it is assumed that, the treatment/exposure period was equivalent to the entire observation period following instillation or lasted until removal of the test material by physiological mechanisms, whichever happened sooner.

Observation period (in vivo):

up to 21 days (ending individually for each animal, when the animal was free of alterations)

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

EYE EVALUATION:

Within 24 hours before treatment start, both eyes of each animal were investigated to ensure that there was no pre-existing corneal damage, iridial inflammation or conjunctival irritation.

One animal was initially treated and, in the absence of a severe effect (no corrosive effect) until 72 hours post instillation, the remaining two animals were committed to the study.

Eyes were evaluated in all animals at approximately 1, 24, 48 and 72 hours after test substance instillation adopting the numerical scoring system listed in Table 1 in the field below. Additional examinations were performed 7, 14 and/or 21 days after the instillation (ending individually for each animal, by the observation time point the animal was entirely free from alterations). Grades attained at 24, 48 and 72 hours after instillation were included in the mean gradings of ocular lesions and used for evaluation of the necessity of eye irritation/corrosion classification. As from 24 h after test substance instillation, staining with fluorescein (Minims Fluorescein Sodium 2 %) supported the recognition of corneal lesions in all animals.

Equipment used for eye evaluation: Otoscope lamp

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks:

, as in one animal (# 2) corneal opacity grade 2 followed at 7, 14 & 21 days after instillation

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks:

, as corneal opacity grade 2 followed at 7, 14 & 21 days after instillation

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

0.1

Max. score:

2

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

2

Reversibility:

other: Iridic changes were not evident

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

2

Reversibility:

other: Iridic changes were not evident

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

0.3

Max. score:

2

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

3.6

Max. score:

4

Reversibility:

not fully reversible within: 21 days in one (#2) of three animals

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

3.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

3.7

Max. score:

4

Reversibility:

not fully reversible within: 21 days, but clear trend to reversibility as on Days 14 & 21 only grade 1

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

3.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

other: Discharge

Basis:

mean

Remarks:

over 3 animals and 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.2

Max. score:

4

Reversibility:

fully reversible within: 21 days

Irritation parameter:

other: Discharge

Basis:

animal #1

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

other: Discharge

Basis:

animal #2

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 21 days

Irritation parameter:

other: Discharge

Basis:

animal #3

Remarks:

mean over 3 time points

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Iridic lesions were not evident throughout the study, except for one incidence of a grade 1 lesion in male #3 at 24 hours after test material instillation (Day 1). Corneal lesions (opacity grade 1) were seen in all animals at 24, 48 and 72 hours after instillation and by Day 7 had entirely disappeared in two animals (#1 and #3), whereas irreversibly had increased in severity to grade 2 in the other animal (#2). Staining with fluorescein solution revealed damage of the anterior corneal epithelium. Although, severity grade-2 corneal opacity remained in animal #2 until Day 21, the affected corneal area gradually decreased from grade 4 on Day 1 to grade 2 on Days 7, 14 and 21. In addition to the findings specified under "Overall irritation/corrosion results", conjunctival redness grade 1 or 2, chemosis grade 2 or 4 and discharge grade 1, 2 or 3, were also seen at 1 h after instillation and, except for chemosis, were fully reversible in all animals. Chemosis was fully reversible in animals #1 and #3, whereas in animal #2 still evident as grade 1 by Day 21. Control eyes (untreated) were without ocular findings throughout the observation period.

Other effects:

Mortality or signs of systemic toxicity were not evident and bodyweight was unaffected by treatment with the test material.

Interpretation of results:

other: Category 1 (irreversible effects on the eye) according to Regulation 1272/2008

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Effects on eye irritation: corrosive

Justification for classification or non-classification

There were only slight and reversible effects of skin irritation in one animal, and no effects in the other two animals. The result of this in vivo test does not necessitate any classification and labelling regarding skin irritation according to REGULATION (EC) 1272/2008.

The irreversible corneal lesions in one animal in the in vivo eye irritation study necessitate classification of WS400402 as “Category 1 (irreversible effects on the eye)" [REGULATION (EC) 1272/2008].