Registration Dossier

Administrative data

Description of key information

The substance is not irritating in the in-vitro assays for skin and eye irritation (OECD 439, OECD 492). In addition, in a study with rabbits (OECD 404), it was slightly irritating to skin, meeting the classification criteria for UN GHS Cat 3.

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:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Red solid
expiry date of sample: 2025
The test item was homogeneous by visual inspection.
Storage at room temperature
pH-value: approx. 8 (moistened with water)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS GmbH, Germany
- Age at study initiation: 4 - 8 months
- Weight at study initiation: 3.25 kg – 4.17 kg
- Housing: Single cages
- Diet ad libitum
- Water ad libitum
- Acclimation period: Acclimatization for at least 5 days before application

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C +/- 3°C
- Humidity (%): 30-70%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

IN-LIFE DATES: From: 6 June 2017 To: 10 July 2017
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 g
Duration of treatment / exposure:
single treatment
Observation period:
14 days
Number of animals:
3
Details on study design:
The cutaneous reactions were assessed immediately after removal of the patch,
approximately 1, 24, 48 and 72 hours after removal of the patch and in weekly intervals
until day 14.
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
0
Remarks on result:
other: no indication of edema at any observation time point
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
1.7
Max. score:
2
Reversibility:
other: fully reversible within 7 and 14 days in two animals, not fully reversed after 14 days in one animal

Animal No. : 1
Exposure periode:  4h   
Readings Erythema Edema Additional Findings
0h * 0 *Test item residues; no erythema could be determined 
1h * 0 *Test item residues; no erythema could be determined 
24h 2 0 -
48h 2 0 -
72h 2 0 -
7d 2 0 -
14d 0 0 SD
Mean
24h - 72h
2.0 0.0  
Animal No. : 2
Exposure periode:  4h   
Readings Erythema Edema Additional Findings
0h * 0 *Test item residues; no erythema could be determined 
1h * 0 *Test item residues; no erythema could be determined 
24h * 0 *Test item residues; no erythema could be determined 
48h 1 0 -
72h 1 0 -
7d 0 0 SD
14d      
Mean
24h - 72h
1.0 0.0  
Animal No. : 3
Exposure periode:  4h   
Readings Erythema Edema Additional Findings
0h * 0 *Test item residues; no erythema could be determined 
1h * 0 *Test item residues; no erythema could be determined 
24h 2 0 *Test item residues
48h 2 0 -
72h 2 0 -
7d 1 0 -
14d 1 0 -
Mean
24h - 72h
2.0 0.0  

  Mean Erythema
24h - 72h
Mean Edema
24h - 72h
 
Animal No. 1 2.0 0.0
Animal No. 2 1.0 0.0
Animal No. 3 2.0 0.0
Mean of
3 Animals
1.7 0.0
Interpretation of results:
Category 3 (mild irritant) based on GHS criteria
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
BASF SE Experimental Toxicology and Ecology 67056 Ludwigshafen, Germany
Species:
human
Strain:
other: not applicable
Details on test animals or tissues and environmental conditions:
not applicable

The EpiOcularTM model (OCL-200) is a three-dimensional non-keratinized tissue construct composed of normal human derived epidermal keratinozytes used to model the human corneal epithelium. The EpiOcularTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm ∅) and are commercially available as kits (EpiOcular™ 200), containing 24 tissues on shipping agarose.

To assess the ability of the test material to directly reduce MTT a pretest was performed.
Vehicle:
unchanged (no vehicle)
Controls:
other: yes (tissue incubations for positive and negative controls included)
Amount / concentration applied:
0.05 mL
Duration of treatment / exposure:
6 hours
Duration of post- treatment incubation (in vitro):
18h
Number of animals or in vitro replicates:
Two tissue samples were used per group.
Details on study design:
Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the testsubstance treated epidermal tissues is compared to that of negative control tissues. The
quotient of the values indicates the relative tissue viability.

As the substance showed a potency for direct reduction of MTT by the test substance, the OD570 values were also measured in freeze-killed control tissues (KC) to calculate the mean OD570 of the test substance treated tissues (mean OD570 KC corrected). Since killed tissue might still have a residual enzyme activity that is able to produce some formazan net OD570 KC is calculated by subtracting the OD570 KC of the NC from the OD570 KC of the test substance. In case the net OD570 KC is greater than zero it is subtracted from the respective mean OD570 to result in the
mean OD570 KC corrected. The mean OD570 KC corrected represents the formazan production linked to the tissue viability and therefore indicates the cytotoxic potency of the test substance.

The color of a test substance may interfere with the color density produced by metabolic capacity of the tissue and would falsify the test results when residues of the test substance remain on the tissues after washing and are extracted by the isopropanol.
Due to the pink to red color of the test substance a pretest was performed as follows: the test substance was applied to killed cell tissues, incubated and removed by washing. Thereafter extraction in isopropanol was performed and the OD570 of the extract was determined spectrophotometrically. Based on the pretest it was judged that no color control (CC) was necessary (details are available with the raw data).
Irritation parameter:
other: viability (%)
Run / experiment:
mean of both tissues
Value:
85.3
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: none

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
Other effects:
Slight red discoloration of the test-substance treated tissues and slight compound residues were observed after the washing procedure.

Table 1: Results

tissue 1 tissue 2 mean Inter-tissue variability (%)
Negative control (water) mean OD570 1.806 1.784 1.795
viability (% of NC) 100.6 99.4 100 1.2
test substance mean OD570 1.535 1.525 1.53
viability (% of NC) 85.5 85 85.3 0.6
Positive control (methyl acetate) mean OD570 0.432 0.486 0.46
viability (% of NC) 24.3 27.1 25.6 2.8
Interpretation of results:
GHS criteria not met
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

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

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. A GLP-compliant OECD 439 study is available for skin irritation. A GLP-compliant OECD 492 study (EpiOcular) is available for eye irritation. In both studies, the scores for the test item treated tissues were below the thresholds for classification as an irritant. As a result the substance is not considered to be classified for skin or eye irritation under Regulation (EC) No. 1272/2008, as amended for the fourteenth time in Regulation (EC) No. 2020/217.