Registration Dossier

Administrative data

Description of key information

Skin: 
(1) In vitro skin irritation assay, reconstructed three dimensional human epidermis model (EpiDermTM), 1 hour: not irritating (Val 1, GLP, OECD 439, BASF 2011)
(2) Rabbit, occlusive, clipped, 1, 5, and 15 min, 20 hours: not irritating (Val 2, non-GLP, Internal standarized test method, BASF 1972).
(3) In vitro skin corrosion assay, reconstructed three dimensional human epidermis model (EpiDermTM), 1 hour: not corrosive (Val 2, non-GLP, OECD 431, BASF 2002)
Eye:
Rabbit: not irritating (Val 2, non-GLP, Internal standarized test method, comparable to OECD 405, BASF 1972).

Key value for chemical safety assessment

Skin irritation / corrosion

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:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study comparable to guideline study (partly limited documentation, e.g. no details about the test substance; 50 µL instead of 100 µL test substance instilled; eye not washed out).
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
(no details about the test substance; 50 µL instead of 100 µL test substance instilled.)
Principles of method if other than guideline:
Internal standardized test method (BASF-Test): 50 µL of the test substance were applied to the conjunctival sac of one eye in each of 2 animals. The animals were examined after 10 min, 1 and 3 h on the day of treatment and up to 8 days afterwards. Findings were recorded daily. The eyes were not washed out after 24 hours as specified in OECD Guideline 405.
GLP compliance:
no
Species:
rabbit
Strain:
Vienna White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: M. Gaukler, Offenbach, Germany
- Weight at study initiation: 2.66 kg (males); 3.12 kg (female)
Vehicle:
unchanged (no vehicle)
Controls:
other: The adjacent eye served as control treated with NaCl.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µL
- Concentration: undiluted

Duration of treatment / exposure:
single application (not washed out)

Observation period (in vivo):
8 days
Number of animals or in vitro replicates:
2 (1 male and 1 female)
Details on study design:
APPLICATION
Single application to the conjunctival sac of the right eyelid.

REMOVAL OF TEST SUBSTANCE
- The test substance was not washed out.

READINGS:
- 24, 48, 72 hours and 8 days after instillation.

SCORING SYSTEM: Draize scoring system, as recommended by the OECD guideline 405

TOOL USED TO ASSESS SCORE: fluorescein (at the end of exposure period)

Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
out of 2 animals
Time point:
other: mean over 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: reversibility: not applicable
Irritation parameter:
iris score
Basis:
mean
Remarks:
out of 2 animals
Time point:
other: mean over 24, 48 and 72 h
Score:
0
Max. score:
2
Reversibility:
other: reversibility: not applicable
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
out of 2 animals
Time point:
other: mean over 24, 48 and 72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 8 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
out of 2 animals
Time point:
other: mean over 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: reversibility: not applicable
Irritant / corrosive response data:
The treatment led to conjunctivae redness with a score of 1 which persisted from 24 to 72 hours and was fully reversible within 8 days in 1/2 animals.
Other effects:
No other findings were observed.

Table 1. Results of eye irritation study.

Rabbit #

 

Time

 

conjunctivae

iris

 

cornea

 

redness

swelling

1

 

 

 

 

1 h

1

0

0

0

24 h

1

0

0

0

48 h

1

0

0

0

72 h

1

0

0

0

8 d

0

0

0

0

average over 24, 48 and 72 h

1.0

0.0

0.0

0.0

2

 

 

 

 

1

0

0

0

0

24 h

0

0

0

0

48 h

0

0

0

0

72 h

0

0

0

0

8 d

0

0

0

0

average over 24, 48 and 72 h

0.0

0.0

0.0

0.0

 

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

There are valid data available for the assessment of the skin and eye irritation potential.

Skin Irritation

The in vitro skin irritation study was performed according to OECD Test Guideline 439 (GLP conditions). A three dimensional human epidermis model (EpiDermTM) was treated with the undiluted test substance for 1 h (irritation test). The skin irritation potential is evaluated by a colourimetric assay (MTT-assay) for measuring the induced cytotoxicity. The test substance is considered irritant if the mean relative tissue viability compared to the negative control tissues is less than or equal to 50%. In the irritation test the mean viability of the test-substance treated tissues determined after an exposure period of 1 hour with about 42 hours post-incubation was 116% (positive control: 6%). Thus, the substance is considered non-irritating in the EpiDerm™ skin irritation test under the test conditions chosen (BASF SE, 61V0587/11A316, 2011). The study is suitable and reliable for assessment of skin irritation as it was performed according to OECD test guideline.

Conclusion: The undiluted test item was not irritating in an in vitro human epidermis model.

A non GLP in vivo skin irritation study according to an internal testing protocol which is generally consistent with the current OECD guideline 404 (BASF SE, XXII/42, 1972) was performed. In brief, 1 mL of the undiluted test substance (analytical purity: unknown) was applied to the clipped, intact skin of two adult male New Zealand White rabbits and covered by an occlusive dressing for the exposure period of 1, 5, and 15 min as well as 20 hours. Except of the 20 hour exposure, the skin was rinsed after removal of test substance at the end of the exposure period and reading of skin reaction was performed 24, and 48 hours as well as 6 days after test item application. Scoring of the findings was done by converting descriptive scores of the raw data to Draize numerical scores. The mean erythema and edema scores for each animal at the reading times 24, and 48 hours were considered for evaluation of the irritating potential according to current criteria.

No signs of irritation were detected at all reading time points. Additionally, no mortality and no signs of intoxication were observed.

Under the experimental conditions chosen, the undiluted test item was not irritating to the skin of rabbits taking into account the "worst case" considerations of the test performance (20h occlusive application of test substance).

An in vitro skin corrosion test was performed similar to OECD Test Guideline 431 (GLP conditions). A three dimensional human epidermis model (EpiDermTM) was treated with the undiluted test substance for 3 min and 1 h. The skin corrosion potential is evaluated by a colourimetric assay (MTT-assay) for measuring the induced cytotoxicity. In the corrosion test the mean viability of the test-substance treated tissues determined after an exposure period of 3 min hour was 99% (positive control: 15%). Thus, the substance is considered non-corrosive in the EpiDerm™ skin corrosion test under the test conditions chosen (BASF SE, 61H0880/002259, 2002). The study is suitable and reliable for assessment of skin corrosion as it was conducted similar to OECD test guideline.

Conclusion: The undiluted test item was not corrosive in an in vitro human epidermis model.

Eye Irritation

A non GLP eye irritation study was performed similar to OECD guideline 405 (1981). In brief, 0.05 mL of the undiluted test substance (purity unknown) was applied into one eye of each of two adult New Zealand White rabbits (BASF SE, XXII/42,1972). The treated eyes were not rinsed after application and reading of eye effects was performed 1, 24, 48 and 72 hours, and for one animal 7 days after test item application. Scoring of the findings was done by converting descriptive scores of the raw data to Draize numerical scores. The mean scores for conjunctivae (redness and chemosis), cornea and iris for each animal at the reading times 24, 48 and 72 hours were considered for evaluation of the irritating potential according to current criteria. Slight conjunctivae redness was seen at reading time points 1 hour (grade 1) up to 72 hours in 1/2 animals, which was reversible within 8 days after application. The average score (24, 48 and 72 hours) for irritation was calculated to be 1.0 (for conjunctivae redness). Slight, reversible conjunctivae redness (grade 1) was noted in the second animal at 3 hours after application. Therefore, average score (24, 48 and 72 hours) for irritation was calculated to be 0.0 (for conjunctivae redness) for the second animal. For corneal opacity, iris and chemosis the average score (24, 48 and 72 hours) for irritation was calculated to be 0.0. The eye irritation study is acceptable (reliability 2), but does not fully satisfy the guideline requirements for an eye irritation test (OECD 405) in rabbits (no rinsing, test substance characterization; reduced test substance bulk - 0.05 mL instead of 0.1 mL; decreased number of animals - 2 animals instead of at least 3 animals).

Under the experimental conditions chosen, the test substance was considered to be not irritating to the eye of rabbits.

This test result was confirmed by two screening assays (BASF SE, 2002): (1) an in vivo assay with only one animal and (2) an in vitro HET-CAM assay both resulted in no indications for an irritating potential of the test substance.

Respiratory system:

There are no specific data available concerning the irritating potential to the respiratory system. Though, exposure of rats to saturated vapors of the test substance did not result in any indications of respiratory tract irritation (see BASF SE, 1972, Inhalation Hazard Assay).

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available experimental test data for skin and eye irritation are reliable and suitable for the purpose of classification under Directive 67/548/EEC. As a result the substance is not warranted to be classified for skin and eye irritation under Directive 67/548/EEC.

 

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data for skin and eye irritation are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. As a result the substance is not warranted to be classified for skin and eye irritation under Regulation (EC) No.1272/2008.