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Description of key information

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

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Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Feb 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
(2002)
Principles of method if other than guideline:
Testing strategy comprised a stepwise approach including the evaluation of existing human and/or animal data showing effects on the skin or mucous membranes, the performance of a SAR evaluation for skin corrosion/irritation, measurement of pH value, the evaluation of data on systemic toxicity vial the dermal route and the performance of a validated in vitro test for skin corrosion (Human 3D Epidermal Skin Model) before in vivo testing for skin irritation/corrosion in rabbits.
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Crl:KBL(NZW)BR
- Sex: female
- Source: Charles River, 88353 Kißleg, Germany
- Age at study initiation: young adult animals
- Weight at study initiation: 3.1-3.2 kg
- Housing: individually in cage units Metall/Noryl by EBECO
- Diet and water: ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3
- Humidity (%): 50 +/- 25
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
other: The surrounding untreated skin served as control
Amount / concentration applied:
0.5 mL
Duration of treatment / exposure:
4 hours
Observation period:
Until changes had completely subsided, however for not more than 14 days after application.
Number of animals:
3
Details on study design:
Due to a possible irritant potential of the test substance, in the first step only one animal was used and three test patches were applied successively to this animal, as described above. The first patch was removed after three minutes. As no serious skin reactions were observed, the second patch was applied and removed after one hour. At this stage the observations indicated that with respect to animal welfare the exposure can be allowed to extend to four hours, therefore the third patch was applied and removed after four hours and the responses were graded one hour later. The test was completed using two additional animals, exposed for four hours.

TEST SITE
On the day before the test, the fur was shorn on the right and left side from the dorso-lateral area of the trunk of each rabbits. 0.5 mL of the pure liquid test substance was applied to the skin of the animal under a gauze patch. The patch was placed on the dorso-lateral areas of the trunk of each animal and was held in place with non-irritant tape for the duration of the exposure period. Thus, access by the animal to the patch and resultant ingestion of the test substance was prevented.
- Area of exposure: approximately 2.5 cm x 2.5 cm

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The exposed skin area was carefully washed with water without altering the existing response, or the integrity of the epidermis.
- Time after start of exposure: after 4 hours

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Remarks:
(redness and eschar formation)
Basis:
animal: #1, #2, #3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal: #1, #2, #3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Other effects:
The control areas did not show any abnormal findings.
There were no systemic intolerance reactions.
Executive summary:

A skin irritation/corrosion test according to OECD 404 was conducted with the test substance. No signs of irritation could be observed at all after 24, 48 and 72 hours postexposure-time (all scores zero). There were no systemic intolerance reactions.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Dec 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
(2004)
GLP compliance:
yes (incl. certificate)
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
The test system EST-1000 (CellSystems, St. Katharinen, Germany) has been validated for skin corrosion and is commercially available.
Vehicle:
unchanged (no vehicle)
Details on test system:
The experiment was carried out on a reconstructed human epidermis EST-1000 (CellSystems, St. Katharinen, Germany). Inserts were of 0.6 cm² size. The model has a functional stratum corneum with an underlying layer of living cells. The barrier function of the stratum corneum is adequate, as has been shown by the supplier.
The corrosive potential of the test item is assessed by determination of its cytotoxic effect on an in vitro reconstructed human epidermis. The test principle is based on the MTT assay reflecting the cell viability after exposure of the epidermal equivalent to topically applied test item.
All tests were performed in triplets for each time point. For testing the inserts were exposed to 50 μL of the undiluted test item for 3 min. (room temperature) and 60 min. (37 +/- 2 °C, 5 % CO2, maximum humidity).
Cell viability was measured by the amount of MTT reduction, i.e. an OD value following exposure to the negative control substance or the test item.
Control samples:
other: as negative control: physiological saline solution (0.9 % NaCl, 50 μL)
Amount/concentration applied:
50 μL per insert
Duration of treatment / exposure:
3 min. (room temperature) and 60 min. (37 °C)
Duration of post-treatment incubation (if applicable):
post-exposure incubation: none
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean of exposure period 3 min.
Value:
91.61
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Remarks on result:
other: no indication of corrosive potential
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean of exposure period 60 min.
Value:
97.91
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Remarks on result:
other: no indication of corrosive potential

 Sample No.

 Test item

 Time [min.]

 % Viability

 1 -3

  control NaCl 0.9 %

 60

 100.00

 4 -6

test substance

 60

 97.91

 10 -12

  control NaCl 0.9 %

 3

 100.00

 13 -15

  test substance

 3

 91.61

The test substance was characterized by no significant impact on cell viability after 3 min. or after the 60 min. period.

Interpretation of results:
other: no corrosive property
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:
March 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
(2002)
Principles of method if other than guideline:
This testing strategy comprised a stepwise approach including the evaluation of existing human and/or animal data showing effects on the eye or the skin, the performance of a SAR evaluation for eye and skin corrosion/irritation, measurement of pH value, the evaluation of data on systemic toxicity via the dermal route, the performance of a validated in vitro test for skin corrosion (Human 3D Epidermal Skin Model) and in vivo testing for skin irritation/corrosion in rabbits before in vivo testing for eye irritation/corrosion in rabbits.
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Strain: Crl:KBL(NZW)BR
- Sex: female
- Source: Charles River, 88353 Kißleg, Germany
- Age at study initiation: young adult animals
- Weight at study initiation: 3.2-3.4 kg
- Housing: individually in cage units Metall/Noryl by EBECO
- Diet and water: ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3
- Humidity (%): 50 +/- 25
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12
Vehicle:
unchanged (no vehicle)
Controls:
other: The other eye, which remained untreated, served as control.
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
The lids were gently held together for about one second in order to prevent loss of the test compound. The eye was rinsed approximately 24 hours following instillation.
Observation period (in vivo):
Animals were monitored after application until the changes had completely subsided, however for not more than 21 days after application.
Number of animals or in vitro replicates:
3
Details on study design:
0.1 ml of the pure liquid test substance was placed into the conjunctival sac of one eye of the first animal after having gently pulIed the lower lid away from the eyeball. The lids were gently held together for about one second in order to prevent loss of the test compound.
Because of the fact that one hour after treatment a severe irritation was not observed, a further two rabbits were treated as described.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The eye was rinsed approximately 24 hours following instillation.

SCORING SYSTEM: Draize
Irritation parameter:
cornea opacity score
Basis:
animal: #1, #2, #3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal: #1, #2, #3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal: #1, #2, #3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal: #1, #2, #3
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Other effects:
The control eyes did not show any abnormal findings.
There were no systemic intolerance reactions.
Executive summary:

An eye irritation/ corrosion test according to OECD 405 was conducted with the test substance. No signs of irritation could be observed at all after 24, 48 and 72 hours postexposure time (all scores zero). There were no systemic intolerance reactions.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The test substance was not irritating to the skin of rabbits (OECD TG 404) and also not irritating to the eye of rabbits (OECD TG 405).

Justification for classification or non-classification

No classification required for skin and eye irritation/corrosion according to Regulation (EC) No 1272/2008, Annex I.