Registration Dossier

Administrative data

Description of key information

Neither classified as a skin or eye irritant

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 09 to March 02, 1987
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study according to OECD Guideline 404 under GLP
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: L'Abbaye de Bellefontaine, F-49122 Begrolles en Mauges, France
- Weight at study initiation: 2.5 ± 0.1 kg
- Housing: Individually in polystyrene cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ±3 °C
- Humidity (%): 50 ± 20
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 ml
Duration of treatment / exposure:
4 hours
Observation period:
1h, 24h, 48 h, and 72 h after exposure termination;
Number of animals:
3 animals
Details on study design:
TEST SITE
- Area of exposure: right flank
- coverage: 6 cm²
- Type of wrap if used: gauze patch held in place with semi-occlusive dressing

SCORING SYSTEM:
Erythema score: 0-4
Edema score: 0-4
Irritation parameter:
erythema score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: 24h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: 48h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: 72h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: 24h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: 48h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: 72h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
Under the conditions of this assay, the test article has no skin irritating potential.
Executive summary:

DBE has been tested for skin irritation on 3 New Zealand White rabbits according to OECD guideline n° 404 and EU guideline n° B.4 (Draize Method) in compliance with Good Laboratory Practice. The rabbits received a dermal dose of 0.5 ml of DBE on a gauze on intact and abraded skin sites under an semiocclusive dressing for a period of 24 hours. Scores for erythema and oedema were measured 1, 24, 48 and 72 hours after patch removal and primary cutaneous index was calculated (maximum possible index =12)

No sign of dermal irritation (score = 0 for erythema and oedema) was observed for intact skin site of the 3 rabbits at any reading time. Primary Irritation Index was = 0.0. No other signs of intoxication were observed.

Based on these results DBE is not classified irritating to skin according to the criteria of the Directive 67/548/CEE and according to EU Regulation 1272/2008 (CLP).

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 09 to March 26, 1987
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study according OECD Guideline 405; Study under GLP Regulations;
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Abbaye de Bellefontaine, F49122 Begrolles en Mauges, France;
- Weight at study initiation: 2.5 ± 0.1 kg
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period:5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 50 ± 20
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml;
Duration of treatment / exposure:
one time application of test article to eye
Observation period (in vivo):
1h after application, followed by daily observation for 3 days (minimum) to 21 days (maximum);
Number of animals or in vitro replicates:
3 animals
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not specified

SCORING SYSTEM: Scoring according to Kay and Calandra (1962) J. Soc. Cosmet. Chem 13, 281-289.
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 1h
Score:
0
Max. score:
0
Reversibility:
fully reversible within: 1d
Remarks on result:
other: 1d
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: 1h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 1d
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: 1h
Score:
3
Max. score:
3
Reversibility:
fully reversible within: 1d
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: 1h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 2d
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: 1h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 2d
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: 1h
Score:
3
Max. score:
3
Reversibility:
fully reversible within: 2d

The observed effects on the conjuctivae and on chemosis 1 hour after application were moderate in two animals and marked in one animal.Two days after the applicaton of the test article, all ocular effects have disappeared.

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
Under the conditions of this study, the test article RP/DE is considered not irritating.
Executive summary:

The eye irritating potential of the test article RP/DE has been investigated in the rabbit under the OECD Guideline 405 and under GLP Regulations. Moderate effects on the conjunctiva and on chemosis were observed in two animals 1 hour after application of the test article while the same effects were marked in one animal. Two days after application of the test article, the effects had disappeared. Under the conditions of this study, the test article RD/DE was considered not irritating to the eye.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

Two in vivo studies using rabbits are available for skin irritation. One study (Clouzeau, 1987) was of reliability 1 according to Klimisch cotation criteria and was therefore selected as the key study. The other study (Sarver, 1989) was of reliability 2 according to Klimisch cotation criteria and was used as a supporting study. In the key study, no signs of skin irritation (erythema or edema) was observed following a 4-hour semi-occlusive application of the test substance to the shaved skin of 3 rabbits and observation up to 72 hours after the end of application. In the supporting study, the skin reactions were very variable from one rabbit to another. At the 72-hour observation timepoint, one rabbit exhibited severe erythema with fissuring of the skin and slight edema, whereas no to mild erythema was observed in the 5 other animals. One rabbit exhibited no dermal irritation at all throughout the observation period. Considering the mean values for erythema :

Rabbit Nr.

Erythema mean score

1

2.3

2

2.0

3

0.0

4

1.7

5

1.0

6

3.3

Global mean value

1.7

 

the incidence of scores of 2 or more was below 2/3 animals, and the global mean value over all the animals tested was lower than 2. Therefore the results of the supporting study are consistent with those of the key study and support the absence of classification for skin irritation.

 

Eye irritation:

Three in vivo studies using rabbits are available for eye irritation:

- In the study selected as a key study based on its reliability 1 according to Klimisch cotation criteria (Clouzeau, 1987), moderate conjunctival and chemosis effects were observed in 2 animals one hour after instillation, and only slight conjunctival redness was noted 24 hours after instillation. All effects were fully reversible within 2 days after application. The intensity and duration of the signs of irritation therefore did not warrant any classification.

- In another study considered of reliability 2 according to Klimisch cotation criteria (Valentine, 1993), signs of ocular irritation were observed in 1 out of the 4 rabbits tested. However, a low volume of instillation (10 µL) was used and the observation period was limited to 48 hours after instillation. No reliable conclusion could be made from this study. However, consistently with the key study, no major sign of eye irritation was observed.

- In the third study, considered of reliability 3 according to Klimisch cotation criteria due to the low number of animals used and the low level of details provided regarding the ocular observations (Silber, 1980), signs of irritation were observed in the unwashed eye of a single rabbit, which may have been susceptible. Such data are therefore inappropriate for classification purpose.

Overall, the results of in vivo testing indicate the absence of a need for classification for eye irritation.


Justification for selection of skin irritation / corrosion endpoint:
GLP and OECD guideline 404 compliant study, with the highest Klimish score (1 vs. 2 for the other one)

Justification for selection of eye irritation endpoint:
GLP and OECD guideline 405 compliant study, with the highest Klimish score (1 vs. 2 or 3 for the other ones)

Justification for classification or non-classification

The incidence and severity of the signs of irritation observed among those different studies were below the criteria of Annex VI Directive 67/548/EEC or EU Regulation 1272/2008 (CLP), therefore warranting no classification for skin or eye irritation.